Biology of blood and marrow transplantation : journal of the American Society for Blood and Marrow Transplantation, Jan 12, 2018
Currently, no consensus has been reached on the optimal blood compartment to be used for surveill... more Currently, no consensus has been reached on the optimal blood compartment to be used for surveillance of cytomegalovirus (CMV) and Epstein-Barr virus (EBV) DNAemia. Although several comparative studies have been performed correlating CMV and EBV DNA loads in whole blood (WB) versus plasma, to our knowledge, no studies to date have analyzed the kinetics of both viruses in the 2 blood compartments. In this retrospective noninterventional multicenter cohort study, the kinetics of CMV and EBV DNA in 121 hematopoietic stem cell transplantation (HSCT) recipients were investigated by analyzing in parallel 569 and 351 paired samples from 80 and 58 sequential episodes of CMV and EBV DNAemia, respectively. Unlike previous studies, this study used a single automated molecular method that was CE-marked and Food and Drug Administration-approved for use in quantifying CMV and EBV DNA in both plasma and WB. Furthermore, the complete viral replication kinetics of all episodes (including both the as...
Aim: To analyze the relation between clinical resistance to ganciclovir (GCV) therapy and isolate... more Aim: To analyze the relation between clinical resistance to ganciclovir (GCV) therapy and isolated cytomegalovirus (CMV) strain and identify the mutation coding GCV resistance in patients after alogeneic hematopoietic stem cell transplantation (HSCT) treated with GCV. Background: Treatment failure is defined as a persistent DNAemia or increasing CMV load after 2 weeks of GCV treatment. CMV resistance to GCV is most frequently caused by mutation in UL 97 gene and develops mostly within 2-3 months after HSCT. Ganciclovir-resistant CMV strain is confirmed by genotype testing. Methods: The group of 68 retrospectively followed patients after HSCT in 2012-2014 (34 males, 34 females, median age 53 years), 40 of them (58.8%) treated with GCV for CMV reactivation with clinical disease. The frequency of blood collection was in noncomplicated course once a weak up to day 100 after HSCT and then according to clinical status of patient. Sequence analysis was performed only in patients with suspected resistence to GCV. CMV viremia was determined by quantitative real-time PCR from full peripheral blood-DNA extraction performed by QIAamp DNA Mini Kit (Qiagen), quantitative DNA determination by CMV RG PCR (Qiagen) with 500 cp/ml as the quantification cut off and 100 cp/ml as the detection cut off. Positive CMV DNA samples were stored at −20 • C for additional testing and determination of mutation time development. The sequence analysis determined UL 97 and UL 54 genes parts, extracted sequences were compared with the reference CMV strain AD169 using the home designed program for detection of differencies by automatical search of mutations and polymorphisms of CMV in UL 97 and UL 54. This program detects the conformity and displays the differences in sequenced DNA. The results were compared with the reference datebase of CMV mutations and polymorphisms. Results: Treatment failure with persistant viraemia occurred in 7 patients. Ganciclovir-resistance based on coding mutation was detected only in 2 patients (mutations L595F and M460I in gene UL 97). Another mutation (N510S in gene UL 97) was found in 1 patient with recurrent CMV replication although he did not meet the criteria for clinical treatment failure. In 5 patients the cause of treatment failure was different. Conclusion: The low incidence of drug resistant CMV isolates in our patients in relatively common clinical treatment failure suggests that slower viral clearance is likely more frequently different from UL 97 and UL 54 genes mutation development.
Little is known about late-onset hemorrhagic cystitis (HC) in children, its relationship to BK vi... more Little is known about late-onset hemorrhagic cystitis (HC) in children, its relationship to BK virus, and treatment with cidofovir (CDV) following hematopoietic stem cell transplantation (HSCT). We prospectively investigated BK virus reactivation in children who underwent HSCT from a matched related donor for thalassemia or sickle cell anemia following busulfan-cyclophosphamide-based conditioning regimens and analyzed risk factors for development of HC and its treatment with CDV. Grade 2-4 HC occurred in 30 patients with a cumulative incidence of 26% (95% confidence interval [CI] 5 18%-34%). The cumulative incidences of BK viruria and viremia were 81% (95% CI 5 69%-89%) and 28% (95% CI 5 18%-40%), respectively. Multivariate analysis revealed that use of antithymocyte globulin (ATG) (hazard ratio [HR] 5 10.5; P 5.001), peak BK viruria .100,000 copies/mL (HR 5 6.2; P 5.004), and grade II-IV acute graft-versus-host disease (HR 5 5.3; P 5 .007) were predictive factors for HC. Nineteen patients with HC were given CDV at 1.5 mg/kg/day 3 times a week, or 5 mg/kg/week. The median duration of therapy was 27 days (range, 21-180 days), and a median of 9 doses were given (range, 6-22). All patients had a complete clinical response (CCR), and 69% had a microbiological response at 4 weeks. Eleven patients with BK virus-related HC receiving supportive care also had CCR. The median duration of HC in these patients was similar to that in patients treated with CDV. None of the patients with HC cleared BK viruria when CCR was achieved. We conclude that late-onset HC is more prevalent in children with sustained high BK viruria who are treated with ATG or who develop graftversus-host disease. Randomized clinical trials are urgently needed to better define the role of CDV in treating BK virus-related HC.
According to World Health Organization (WHO)-defined criteria, patients presenting clinically as ... more According to World Health Organization (WHO)-defined criteria, patients presenting clinically as essential thrombocythemia (ET) may show early primary myelofibrosis (PMF) with accompanying thrombocythemia [1]. Previous clinicopathological studies revealed that laboratory parameters like gender-matched hemoglobin (Hb), white blood cell (WBC) count, and particularly lactate dehydrogenase (LDH) values are significantly different in PMF [2]. By strictly applying the WHO criteria, our investigation was aimed to study sensitivity and specificity of these features in an exploratory cohort of 536 patients and to validate the results on an independently recruited series of 321 strictly corresponding patients. The discriminatory power of these parameters (Hb, WBC, and LDH) was tested by plotting their receiver operating characteristic curves. The best performance was found for LDH (areas under the curve, AUC 5 0.7059). WBC and Hb had superimposable curves, with AUC of 0.6279 and 0.6257, respectively. A diagnostic algorithm was generated by applying these parameters in a stepwise fashion. Nearly half of the patients could be correctly allocated to WHOdefined ET or early PMF in both cohorts investigated. It is important to note that this result does not substitute bone marrow morphology with hematological parameters, however, in clinical practice may alert physicians to get more suspicious of early PMF in a patient presumably presenting with ET. Following current criteria established by the World Health Organization (WHO), patients presenting with a clinical picture of essential thrombocythemia (ET) can actually have an early primary myelofibrosis (PMF) [3]. Incidences may vary depending on the accuracy of applied diagnostic guidelines [4][5][6] and was recently reported in about 18% of cases of socalled ET [7]. Laboratory tests which are significantly different in early PMF as compared with histologically confirmed ET (WHO-ET) include decreased Hb, increased white blood cell (WBC) and lactate dehydrogenase (LDH) values. Although bone marrow biopsy represents the gold standard for differentiating WHO-ET from early PMF [8], in clinical practice it might be very useful to know whether one or more of these baseline laboratory parameters may help to differentiate these two entities. To tackle this issue, we evaluated sensitivity (SE) and specificity (SP) of blood cells counts and LDH for the diagnosis of early PMF versus WHO-ET in an exploratory cohort of 536 histologically diagnosed patients aimed to propose an algorithm for the practical clinical use of these tests. This algorithm was then applied to an independently recruited validation cohort of 321 patients presenting with similar diagnosis of early PMF versus WHO-ET. The starting point of this study was an international database of 1,071 patients with ET either confirmed by WHO criteria (891 cases) or revised to early PMF (180 cases) as detailed elsewhere [7]. From this database, 536 patients (50%) who had complete laboratory data measured at diagnosis were extracted and constituted the training set of our study. Clinical and laboratory characteristics of the 536 selected patients were not different from that of the initial population. To recognize early PMF, the salient parameters SE and SP have been evaluated by receiver operating characteristic (ROC) curves (Fig. ). The worst performance was registered for platelets (PLT) count: its areas under the curve (AUC) was only 0.5628, not significantly different from the reference value of 0.50 (P 5 0.154). Thresholds of Hb, WBC and LDH were searched to achieve at least 90% of SE or SP. Table shows that Hb < 12 g/dL for women or <13 g/dL for men, or WBC >513 3 10 9 /L had higher SP (92 and 91%, respectively). High SP of these two tests is associated with few false positives (WHO-ET cases, incorrectly identified as early PMF), thus it is highly related to the presence of early PMF. Concerning high levels of LDH, we could not find an upper threshold with a SP >90% for diagnosis of early PMF and including at least 5% of patients. On the contrary, LDH < 200 mU/mL and WBC < 7 3 10 9 /L had good SE (91 and 94%, respectively). High SE produces few false negatives (early PMF cases not recognized and incorrectly classified as ET), thus it is highly related to the confirm of WHO-ET diagnosis. To optimize the use of this data in daily clinical practice, a step-by-step algorithm was proposed. By applying the criterium ''anemia,'' SP was 92% indicating that 46/536 (9%) of patients presenting anemia were correctly classified in 92% of cases as early PMF. In the 490 remaining patients, we applied two different WBC cut-offs: 97 (18%) and 43 (8%) patients were classified as WHO-ET or early PMF. Finally, in the remaining patients, LDH value (<200 mU/mL) classified as WHO-ET in another 13% of patients. Thus, following these steps, nearly half of patients (48%) could be classified as WHO-ET (by SE) or early PMF (by SP), assuming at each step a margin of error of about 10% (Fig. ). For the remaining 50% of patients, laboratory results did not allow to suspect or exclude the presence of early PMF. To validate these results, the performance of the same thresholds of laboratory parameters was tested in 321 patients classified as WHO-ET or early PMF (Cologne cohort). SP of anemia was 84%, WBC below 7 3 10 9 /L or above 13 3 10 9 /L had 91 and 81% of SE and SP, respectively. LDH values < 200 revealed a SE of 85%. By applying the above reported flow chart, the percentage of patients classified as WHO-ET or early PMF was almost similar (46%), even though the margin of error at each step was higher, likely due to smaller number of patients. In conclusion, while patients presenting clinically with ET can now be discriminated as true ET or early PMF by adopting the WHO 2008 criteria that require bone marrow histology, considering Hb, WBC, and LDH, this differentiation can be recognized in about 50% of patients with a good approximation. On the other hand, for a definitive proof, bone marrow histology remains still an integral part for final diagnosis. For this reason, it should be emphasized that results of our study do not substitute bone marrow biopsy with laboratory parameters to distinguish true ET from early PMF. Instead, it is to provide clinicians with laboratory parameters that should increase suspicion of early PMF in a patient with a working clinical diagnosis of ET. Laboratory parameters considered in this study were those already found significantly different between early PMF and WHO-ET: Hb (g/dL), WBC (310 9 /L), PLT (310 9 /L), and LDH (mU/mL). The discriminatory ability of these parameters in correctly classifying patients in the early PMF or WHO-ET groups was initially tested by plotting their ROC curves and comparing the relative AUC with the value of 0.50 (which stands for the completely useless application of the test) [9,10]. Three parameters with statistically significant discriminatory power were chosen (Hb, WBC, and LDH) and opportune thresholds searched to guarantee at least 90% of SE or SP [11]. Finally, a diagnostic algorithm was designed. The validation set of this analysis was constituted by 321 patients with WHO-ET (n 5 62) or early PMF (n 5 259) diagnosed by the same pathologist who confirmed the training set cohort and collected in the Institute for Pathology,
Almost all cervical cancers are human papillomavirus (HPV)-positive. Some aspects of HPV carcinog... more Almost all cervical cancers are human papillomavirus (HPV)-positive. Some aspects of HPV carcinogenesis, such as factors involved in the transformation process and the mono-or polyclonal origin of the carcinogenic process, need to be defined. The latter aspect is addressed in our study. Cervical samples were collected from 102 patients with squamous cell carcinoma. The HPV positivity was established by PCR analysis performed using consensus and specific primers for the L1 and E6/E7 regions, respectively. Eightyseven samples were positive for the L1 gene and 5 for the E6/E7 genes. Overall, 92 samples contained segments of HPV-DNA (90.2%). HPV-16 was most frequently found either alone or associated with other genotypes (63%). All genotypes identified as a single infection, except HPV-73, belonged to the high-risk HPV group. Among multiple infections, the HPV-31+54 couple was the most frequent. The presence of two genotypes in a primary tumor raises the question of their distribution in a single tumor cell. We attempted to answer this question by comparing the HPV patterns in primary tumors and metastases, considering that metastases derive from cell clones released from the primary tumor. The HPV patterns of primary tumors and metastases overlapped in most patients, even when primary tumors contained a double genotype, thus suggesting that single tumor cells may contain multiple HPV genotypes.
M e d i c a 241 dei neonati screenati è risultata inferiore a quella registrata su territorio naz... more M e d i c a 241 dei neonati screenati è risultata inferiore a quella registrata su territorio nazionale (1,61%0). Questo dato merita approfondimento poiché sembra non rispecchiare la realtà della intera regione: su 31 casi di neonati, nati nello stesso periodo dello studio, ospedalizzati in altri nosocomi regionali e giunti alla nostra attenzione, 10 sono risultati affetti da infezione congenita da CMV (dati in pubblicazione). Per quanto riguarda invece la stima del ruolo dell'infezione congenita da CMV nell'eziologia della SNS, la frequenza è risultata pari al 52.9% relativamente alla totalità dei bambini sordi esaminati (9/17); di questi 4 erano senza causa di sordità o fattori di rischio noti, 4 presentavano sintomatologia severa per CMV (SGA, calcificazioni cerebrali, dilatazione ventricolare, ipotonia) e 1 sintomatologia misconosciuta per CMV.
Accurate determination of hepatitis C virus RNA level is essential for evaluating the response to... more Accurate determination of hepatitis C virus RNA level is essential for evaluating the response to antiviral therapy, to determine the duration of treatment, and to predict treatment outcome. Currently, two real-time based polymerase chain reaction assays are used widely to monitor the hepatitis C RNA level: the Abbott RealTime HCV assay and the Cobas Taqman HCV assay. Recently, a third assay has become commercially available: the Artus HCV QS-RGQ assay, which uses the QIAsymphony SP/AS platform for sample preparation and PCR-setup, and the Rotor-Gene Q for amplification and detection. In this study, the performance of the Artus HCV QS-RGQ assay was tested on 105 plasma samples and compared to that of the Cobas Taqman HCV assay. Linear regression analysis showed a good agreement between the two assays. A slightly better sensitivity was observed with the Cobas Taqman assay, while higher hepatitis C viral RNA levels were measured by the Artus HCV QS-RGQ assay in samples positive for hepatitis C genotypes 4. Taken together, the data suggest that the Artus HCV QS-RGQ assay is useful in a diagnostic setting. The combination with the versatile QIAsymphony SP/AS system may represent a major advantage for clinical virological laboratories aiming at optimizing their workflow.
HIV-1 viral load determination is a crucial step for monitoring the efficacy of highly active ant... more HIV-1 viral load determination is a crucial step for monitoring the efficacy of highly active antiretroviral therapy (HAART) and predicts disease progression. Real-time PCR based assays are available for monitoring the viral load. They differ in sensitivity, genomic target region and dynamic range. In this study, the performance of the Roche Cobas Taqman HIV-1 v2.0 was evaluated on plasma samples from HIV-1 positive patients in parallel with the Abbott RealTime HIV-1 assay in a routine diagnostic setting. Overall, there was a good agreement between the two assays. However, some samples detected by the Abbott RealTime HIV-1 assay but below the limit of quantitation of the assay were found negative result when tested with the Roche Cobas Taqman HIV-1 v2.0. It is conceivable that signal anomalies or background noise may affect the lower-end precision of the Abbott RealTime HIV-1 assay. Based on these results, it is concluded that it is not recommended to switch platform during longitudinal viral load monitoring of HIV-1 positive patients.
International Journal of Immunopathology and Pharmacology, Oct 1, 2012
Human papillomavirus (HPV) is the most common sexually transmitted agent worldwide. Prevalence va... more Human papillomavirus (HPV) is the most common sexually transmitted agent worldwide. Prevalence varies according to the geographic regions, and is highest in developing countries. Geographic differences exist also in the detection rate of oncogenic types in malignant cervical lesions. In this study, the prevalence of HPV infection as well as the spectrum of HPV types was evaluated in Italian and immigrant women of the urban area of Rome. Several risk factors (age at first intercourse, number of partners, smoking, pregnancy, age at first pregnancy, contraception, education, and menarche) were taken into consideration. Overall, there was a high prevalence of HPV infection in the two groups studied. No significant differences were observed in the spectrum ofHPV types detected. HPV 16 and 18 were the types detected more frequently in both groups. Interestingly, HPV 54 and 70 were found only in the immigrants. Whether this finding reflects a recent introduction of these HPV types in the population studied remains to be established. Monitoring of HPV types in the population is advisable, especially in countries like Italy which is a destination and a gateway for immigrants directed towards north and central Europe. The introduction of high risk HPV variants may have a clinical impact and affect the diagnostic procedures.
Objective: Oncogenic human papillomaviruses (HPVs) are the etiological agents of cervical cancer.... more Objective: Oncogenic human papillomaviruses (HPVs) are the etiological agents of cervical cancer. Different cofactors might be needed for malignant transformation, but they still remain elusive. Methods: To delineate the role of Chlamydia trachomatis (CT) and herpes simplex virus type 2 (HSV2) in HPV-positive cervical intraepithelial neoplasia (CIN) lesions and cervical carcinoma a series of 149 cervical cancer and CIN biopsies were analyzed for CT and HSV2 DNA by PCR, and HPV genotyped by InnoLipa. Monitoring of aberrations in key intracellular pathways due to CT/HSV2 and HPV co-expression were analyzed with 13 biomarkers. Results: Of the 149 samples tested, 136 were HPV DNA positive; 32/136 contained also CT DNA and 29 HSV2 DNA. Detection of CT was significantly (p = 0.0001) related to multiple-type HPV infections, while HSV2 was of borderline significance (p = 0.053). Of the 13 biomarkers tested, cytoplasmic and nuclear NF-ĸB and VEGF-C were significantly increased in CT+/HPV+ lesions; p = 0.023, p = 0.045, and p = 0.020 as well as survivin, p = 0.026. Survivin was the only marker that was overexpressed also in HSV2+/HPV+ lesions, p = 0.027. Conclusions: CT infection favors the entry and persistence of multiple HR-HPV types, which leads to viral integration, inhibition of apoptosis, overexpression of E6/E7 oncogenes and cell transformation.
Background: In the last two years, the SARS-CoV-2 pandemic has determined radical changes in huma... more Background: In the last two years, the SARS-CoV-2 pandemic has determined radical changes in human behaviors and lifestyles, with a drastic reduction in socialization due to physical distancing and self-isolation. These changes have also been reflected in the epidemiological patterns of common respiratory viruses. For this reason, early discrimination of respiratory viruses is important as new variants emerge. Methods: Nasopharyngeal swabs of 2554 patients, with clinically suspected Acute Respiratory Infections (ARIs) from October 2019 to November 2021, were collected to detect 1 or more of the 23 common respiratory pathogens, especially viruses, via BioFilmArray RP2.1plus, including SARS-CoV-2. Demographical characteristics and epidemiological analyses were performed as well as a laboratory features profile of positive patients. Results: An observational study on 2300 patients (254 patients were excluded because of missing data) including 1560 men and 760 women, median age of 64.5 ...
E-cadherin plays a pivotal role in maintenance of normal adhesion in epithelial cells but has als... more E-cadherin plays a pivotal role in maintenance of normal adhesion in epithelial cells but has also been shown to suppress tumour invasion and participate in cell signalling. Known to be capable of reversing the invasive phenotype of high-risk human papillomavirus (HPV)-transformed keratinocytes, E-cadherin is down-regulated in CIN and cervical cancer (CC), but still incompletely studied as an intermediate endpoint marker in this disease. As part of our HPV-PathogenISS study, a series of 150 CCs and 152 CIN lesions were examined using immunohistochemical (IHC) staining for E-cadherin, and tested for HPV using PCR with three primer sets (MY09/11, GP5+/GP6, SPF). Follow-up data were available from all squamous cell carcinoma (SCC) patients, and 67 CIN lesions were monitored with serial PCR for HPV after cone treatment. Expression of E-cadherin was reduced in parallel with the increasing grade of CIN, with major down-regulation upon transition to CIN3 and further to invasive cancer (OR ...
In women with HIV-associated immunosuppression, HPV infections have an increased risk of progress... more In women with HIV-associated immunosuppression, HPV infections have an increased risk of progression to high-grade cervical intraepithelial neoplasia (CIN). With the HAART-induced prolonged survival and more protracted clinical course of AIDS, progression of CIN to cervical cancer (CC) has become a clinically relevant issue, and the mechanisms responsible for HIV-HPV interactions need further elucidation. The study design and analysis of the baseline data of our new project are presented. This project is a combination of a prospective cohort study of HIV- and HIV+ women, and a retrospective analysis of CIN lesions and cervical cancer. Up to the present, 244 women have been enrolled (17 HIV+) and subjected to epidemiological interview, colposcopic examination, sampling for HPV testing and typing (PCR, InnoLiPA), and HPV serology. The retrospective series of biopsies were analysed for 13 biomarkers (monitoring key molecular events) using immunohistochemistry and tested for HPV by PCR ...
Several detailed algorithms for the appropriate use of human papillomavirus (HPV) testing in the ... more Several detailed algorithms for the appropriate use of human papillomavirus (HPV) testing in the management of women with abnormal Pap (Papanicolaou) smears have been launched, but their direct country-to-country adoption is difficult. This necessitates their testing in individual settings, which is ongoing in our colposcopy referral clinic. A series of 224 consecutive women attending the clinic with the usual referral indications (ASC-US or higher in Pap) were examined by the conventional diagnostic tools (PAP smear, colposcopy, punch biopsy) and subjected to HPV testing and viral typing for both low-risk (L-R) and high-risk (H-R) types by nested PCR-based techniques. Predictors of the high-grade diagnostic categories were analysed using both univariate- and multivariate modelling, and the performance characteristics (sensitivity, specificity, NPV, PPV) of all tests in detecting high-grade CIN were calculated. In the PAP test, ASC-US smears were most common (37.9%), followed by low...
In recent years, efforts have been made to identify molecular markers as potential screening tool... more In recent years, efforts have been made to identify molecular markers as potential screening tools in the early detection of cervical cancer precursors. One-hundred-eighty-two women admitted to the Colposcopy Unit of Tor Vergata University Hospital were enrolled in this study. The inclusion criteria were: i) Pap test with atypical squamous cells of undetermined significance (ASCUS) or low-grade squamous intraepithelial lesion (LSIL); ii) normal cytology but human papillomavirus (HPV) DNA test positive for at least one of the most frequently detected five high-risk HPV types (16, 18, 31, 33 and 45). HPV DNA was detected with the HPV Sign kit and the type was assigned by pyrosequencing using the PyroMark ID System. E6/E7 transcripts of the high-risk HPV types 16, 18, 31, 33 and 45 were detected by the NucliSense EasyQ HPV kit. Overall, 90 (49.5%) patients were HPV-DNA negative, whereas 92 (50.5%) were HPV-DNA positive. Single infections were detected in 55 women: HPV 16 ranked first (...
Novel respiratory viruses have been identified as possible agents of upper and lower respiratory ... more Novel respiratory viruses have been identified as possible agents of upper and lower respiratory tract infections. Multiplex real-time PCRs have been developed to identify clinically relevant respiratory pathogens. In this study, 178 respiratory samples already screened for influenza virus types A and B by Flu A/B ASR real-time PCR kit were retrospectively analyzed with the Respiratory Multi Well System (MWS) r-gene™ real-time PCR kit which detects a wide spectrum of respiratory pathogens. The goal was to demonstrate the importance of a wide spectrum screening compared to a single diagnostic request. The Flu A/B ASR kit detected influenza B virus in 1.7% of the samples (3/178) and no influenza A virus. The MWS r-gene™ kit detected influenza virus in 6.7% (12/178) of samples (0.6% influenza A, and 6.2% influenza B), while the overall detection rate for respiratory pathogens was 54% (96/178). Co-infections were detected in 8/178 (4.5%) samples. Adenovirus was the infectious agent dete...
Anyplex™II HPV28 is a new PCR assay designed for HPV genotyping. It can detect 28 HPV types inclu... more Anyplex™II HPV28 is a new PCR assay designed for HPV genotyping. It can detect 28 HPV types including 19 high-risk and 9 low-risk types. This study evaluated the performance of Anyplex™II HPV28 on 123 fresh cervical samples screened in parallel with HPV Sign® Genotyping Test. Of the 123 samples screened, 93 were positive, 15 negative, and 15 discordant. The total number of HPV positive samples combined was 108: 38 single infections and 70 multiple infections. The agreement between the two tests was 87.8%, κ=0.592. Genotype specific agreement was strong for HPV 16 (k=0.761), HPV 18 (k=0.674), and HPV 35 (k=0.796). Sensitivity and specificity of Anyplex™II HPV28 assay using HPV Sign® Genotyping Test as reference was 84.8% and 94%; conversely, sensitivity and specificity of HPV Sign® Genotyping Test was 29% and 99.5%. Anyplex™II HPV28 assay is a sensitive and specific assay suitable for HPV genotyping but requires clinical validation.
Biology of blood and marrow transplantation : journal of the American Society for Blood and Marrow Transplantation, Jan 12, 2018
Currently, no consensus has been reached on the optimal blood compartment to be used for surveill... more Currently, no consensus has been reached on the optimal blood compartment to be used for surveillance of cytomegalovirus (CMV) and Epstein-Barr virus (EBV) DNAemia. Although several comparative studies have been performed correlating CMV and EBV DNA loads in whole blood (WB) versus plasma, to our knowledge, no studies to date have analyzed the kinetics of both viruses in the 2 blood compartments. In this retrospective noninterventional multicenter cohort study, the kinetics of CMV and EBV DNA in 121 hematopoietic stem cell transplantation (HSCT) recipients were investigated by analyzing in parallel 569 and 351 paired samples from 80 and 58 sequential episodes of CMV and EBV DNAemia, respectively. Unlike previous studies, this study used a single automated molecular method that was CE-marked and Food and Drug Administration-approved for use in quantifying CMV and EBV DNA in both plasma and WB. Furthermore, the complete viral replication kinetics of all episodes (including both the as...
Aim: To analyze the relation between clinical resistance to ganciclovir (GCV) therapy and isolate... more Aim: To analyze the relation between clinical resistance to ganciclovir (GCV) therapy and isolated cytomegalovirus (CMV) strain and identify the mutation coding GCV resistance in patients after alogeneic hematopoietic stem cell transplantation (HSCT) treated with GCV. Background: Treatment failure is defined as a persistent DNAemia or increasing CMV load after 2 weeks of GCV treatment. CMV resistance to GCV is most frequently caused by mutation in UL 97 gene and develops mostly within 2-3 months after HSCT. Ganciclovir-resistant CMV strain is confirmed by genotype testing. Methods: The group of 68 retrospectively followed patients after HSCT in 2012-2014 (34 males, 34 females, median age 53 years), 40 of them (58.8%) treated with GCV for CMV reactivation with clinical disease. The frequency of blood collection was in noncomplicated course once a weak up to day 100 after HSCT and then according to clinical status of patient. Sequence analysis was performed only in patients with suspected resistence to GCV. CMV viremia was determined by quantitative real-time PCR from full peripheral blood-DNA extraction performed by QIAamp DNA Mini Kit (Qiagen), quantitative DNA determination by CMV RG PCR (Qiagen) with 500 cp/ml as the quantification cut off and 100 cp/ml as the detection cut off. Positive CMV DNA samples were stored at −20 • C for additional testing and determination of mutation time development. The sequence analysis determined UL 97 and UL 54 genes parts, extracted sequences were compared with the reference CMV strain AD169 using the home designed program for detection of differencies by automatical search of mutations and polymorphisms of CMV in UL 97 and UL 54. This program detects the conformity and displays the differences in sequenced DNA. The results were compared with the reference datebase of CMV mutations and polymorphisms. Results: Treatment failure with persistant viraemia occurred in 7 patients. Ganciclovir-resistance based on coding mutation was detected only in 2 patients (mutations L595F and M460I in gene UL 97). Another mutation (N510S in gene UL 97) was found in 1 patient with recurrent CMV replication although he did not meet the criteria for clinical treatment failure. In 5 patients the cause of treatment failure was different. Conclusion: The low incidence of drug resistant CMV isolates in our patients in relatively common clinical treatment failure suggests that slower viral clearance is likely more frequently different from UL 97 and UL 54 genes mutation development.
Little is known about late-onset hemorrhagic cystitis (HC) in children, its relationship to BK vi... more Little is known about late-onset hemorrhagic cystitis (HC) in children, its relationship to BK virus, and treatment with cidofovir (CDV) following hematopoietic stem cell transplantation (HSCT). We prospectively investigated BK virus reactivation in children who underwent HSCT from a matched related donor for thalassemia or sickle cell anemia following busulfan-cyclophosphamide-based conditioning regimens and analyzed risk factors for development of HC and its treatment with CDV. Grade 2-4 HC occurred in 30 patients with a cumulative incidence of 26% (95% confidence interval [CI] 5 18%-34%). The cumulative incidences of BK viruria and viremia were 81% (95% CI 5 69%-89%) and 28% (95% CI 5 18%-40%), respectively. Multivariate analysis revealed that use of antithymocyte globulin (ATG) (hazard ratio [HR] 5 10.5; P 5.001), peak BK viruria .100,000 copies/mL (HR 5 6.2; P 5.004), and grade II-IV acute graft-versus-host disease (HR 5 5.3; P 5 .007) were predictive factors for HC. Nineteen patients with HC were given CDV at 1.5 mg/kg/day 3 times a week, or 5 mg/kg/week. The median duration of therapy was 27 days (range, 21-180 days), and a median of 9 doses were given (range, 6-22). All patients had a complete clinical response (CCR), and 69% had a microbiological response at 4 weeks. Eleven patients with BK virus-related HC receiving supportive care also had CCR. The median duration of HC in these patients was similar to that in patients treated with CDV. None of the patients with HC cleared BK viruria when CCR was achieved. We conclude that late-onset HC is more prevalent in children with sustained high BK viruria who are treated with ATG or who develop graftversus-host disease. Randomized clinical trials are urgently needed to better define the role of CDV in treating BK virus-related HC.
According to World Health Organization (WHO)-defined criteria, patients presenting clinically as ... more According to World Health Organization (WHO)-defined criteria, patients presenting clinically as essential thrombocythemia (ET) may show early primary myelofibrosis (PMF) with accompanying thrombocythemia [1]. Previous clinicopathological studies revealed that laboratory parameters like gender-matched hemoglobin (Hb), white blood cell (WBC) count, and particularly lactate dehydrogenase (LDH) values are significantly different in PMF [2]. By strictly applying the WHO criteria, our investigation was aimed to study sensitivity and specificity of these features in an exploratory cohort of 536 patients and to validate the results on an independently recruited series of 321 strictly corresponding patients. The discriminatory power of these parameters (Hb, WBC, and LDH) was tested by plotting their receiver operating characteristic curves. The best performance was found for LDH (areas under the curve, AUC 5 0.7059). WBC and Hb had superimposable curves, with AUC of 0.6279 and 0.6257, respectively. A diagnostic algorithm was generated by applying these parameters in a stepwise fashion. Nearly half of the patients could be correctly allocated to WHOdefined ET or early PMF in both cohorts investigated. It is important to note that this result does not substitute bone marrow morphology with hematological parameters, however, in clinical practice may alert physicians to get more suspicious of early PMF in a patient presumably presenting with ET. Following current criteria established by the World Health Organization (WHO), patients presenting with a clinical picture of essential thrombocythemia (ET) can actually have an early primary myelofibrosis (PMF) [3]. Incidences may vary depending on the accuracy of applied diagnostic guidelines [4][5][6] and was recently reported in about 18% of cases of socalled ET [7]. Laboratory tests which are significantly different in early PMF as compared with histologically confirmed ET (WHO-ET) include decreased Hb, increased white blood cell (WBC) and lactate dehydrogenase (LDH) values. Although bone marrow biopsy represents the gold standard for differentiating WHO-ET from early PMF [8], in clinical practice it might be very useful to know whether one or more of these baseline laboratory parameters may help to differentiate these two entities. To tackle this issue, we evaluated sensitivity (SE) and specificity (SP) of blood cells counts and LDH for the diagnosis of early PMF versus WHO-ET in an exploratory cohort of 536 histologically diagnosed patients aimed to propose an algorithm for the practical clinical use of these tests. This algorithm was then applied to an independently recruited validation cohort of 321 patients presenting with similar diagnosis of early PMF versus WHO-ET. The starting point of this study was an international database of 1,071 patients with ET either confirmed by WHO criteria (891 cases) or revised to early PMF (180 cases) as detailed elsewhere [7]. From this database, 536 patients (50%) who had complete laboratory data measured at diagnosis were extracted and constituted the training set of our study. Clinical and laboratory characteristics of the 536 selected patients were not different from that of the initial population. To recognize early PMF, the salient parameters SE and SP have been evaluated by receiver operating characteristic (ROC) curves (Fig. ). The worst performance was registered for platelets (PLT) count: its areas under the curve (AUC) was only 0.5628, not significantly different from the reference value of 0.50 (P 5 0.154). Thresholds of Hb, WBC and LDH were searched to achieve at least 90% of SE or SP. Table shows that Hb < 12 g/dL for women or <13 g/dL for men, or WBC >513 3 10 9 /L had higher SP (92 and 91%, respectively). High SP of these two tests is associated with few false positives (WHO-ET cases, incorrectly identified as early PMF), thus it is highly related to the presence of early PMF. Concerning high levels of LDH, we could not find an upper threshold with a SP >90% for diagnosis of early PMF and including at least 5% of patients. On the contrary, LDH < 200 mU/mL and WBC < 7 3 10 9 /L had good SE (91 and 94%, respectively). High SE produces few false negatives (early PMF cases not recognized and incorrectly classified as ET), thus it is highly related to the confirm of WHO-ET diagnosis. To optimize the use of this data in daily clinical practice, a step-by-step algorithm was proposed. By applying the criterium ''anemia,'' SP was 92% indicating that 46/536 (9%) of patients presenting anemia were correctly classified in 92% of cases as early PMF. In the 490 remaining patients, we applied two different WBC cut-offs: 97 (18%) and 43 (8%) patients were classified as WHO-ET or early PMF. Finally, in the remaining patients, LDH value (<200 mU/mL) classified as WHO-ET in another 13% of patients. Thus, following these steps, nearly half of patients (48%) could be classified as WHO-ET (by SE) or early PMF (by SP), assuming at each step a margin of error of about 10% (Fig. ). For the remaining 50% of patients, laboratory results did not allow to suspect or exclude the presence of early PMF. To validate these results, the performance of the same thresholds of laboratory parameters was tested in 321 patients classified as WHO-ET or early PMF (Cologne cohort). SP of anemia was 84%, WBC below 7 3 10 9 /L or above 13 3 10 9 /L had 91 and 81% of SE and SP, respectively. LDH values < 200 revealed a SE of 85%. By applying the above reported flow chart, the percentage of patients classified as WHO-ET or early PMF was almost similar (46%), even though the margin of error at each step was higher, likely due to smaller number of patients. In conclusion, while patients presenting clinically with ET can now be discriminated as true ET or early PMF by adopting the WHO 2008 criteria that require bone marrow histology, considering Hb, WBC, and LDH, this differentiation can be recognized in about 50% of patients with a good approximation. On the other hand, for a definitive proof, bone marrow histology remains still an integral part for final diagnosis. For this reason, it should be emphasized that results of our study do not substitute bone marrow biopsy with laboratory parameters to distinguish true ET from early PMF. Instead, it is to provide clinicians with laboratory parameters that should increase suspicion of early PMF in a patient with a working clinical diagnosis of ET. Laboratory parameters considered in this study were those already found significantly different between early PMF and WHO-ET: Hb (g/dL), WBC (310 9 /L), PLT (310 9 /L), and LDH (mU/mL). The discriminatory ability of these parameters in correctly classifying patients in the early PMF or WHO-ET groups was initially tested by plotting their ROC curves and comparing the relative AUC with the value of 0.50 (which stands for the completely useless application of the test) [9,10]. Three parameters with statistically significant discriminatory power were chosen (Hb, WBC, and LDH) and opportune thresholds searched to guarantee at least 90% of SE or SP [11]. Finally, a diagnostic algorithm was designed. The validation set of this analysis was constituted by 321 patients with WHO-ET (n 5 62) or early PMF (n 5 259) diagnosed by the same pathologist who confirmed the training set cohort and collected in the Institute for Pathology,
Almost all cervical cancers are human papillomavirus (HPV)-positive. Some aspects of HPV carcinog... more Almost all cervical cancers are human papillomavirus (HPV)-positive. Some aspects of HPV carcinogenesis, such as factors involved in the transformation process and the mono-or polyclonal origin of the carcinogenic process, need to be defined. The latter aspect is addressed in our study. Cervical samples were collected from 102 patients with squamous cell carcinoma. The HPV positivity was established by PCR analysis performed using consensus and specific primers for the L1 and E6/E7 regions, respectively. Eightyseven samples were positive for the L1 gene and 5 for the E6/E7 genes. Overall, 92 samples contained segments of HPV-DNA (90.2%). HPV-16 was most frequently found either alone or associated with other genotypes (63%). All genotypes identified as a single infection, except HPV-73, belonged to the high-risk HPV group. Among multiple infections, the HPV-31+54 couple was the most frequent. The presence of two genotypes in a primary tumor raises the question of their distribution in a single tumor cell. We attempted to answer this question by comparing the HPV patterns in primary tumors and metastases, considering that metastases derive from cell clones released from the primary tumor. The HPV patterns of primary tumors and metastases overlapped in most patients, even when primary tumors contained a double genotype, thus suggesting that single tumor cells may contain multiple HPV genotypes.
M e d i c a 241 dei neonati screenati è risultata inferiore a quella registrata su territorio naz... more M e d i c a 241 dei neonati screenati è risultata inferiore a quella registrata su territorio nazionale (1,61%0). Questo dato merita approfondimento poiché sembra non rispecchiare la realtà della intera regione: su 31 casi di neonati, nati nello stesso periodo dello studio, ospedalizzati in altri nosocomi regionali e giunti alla nostra attenzione, 10 sono risultati affetti da infezione congenita da CMV (dati in pubblicazione). Per quanto riguarda invece la stima del ruolo dell'infezione congenita da CMV nell'eziologia della SNS, la frequenza è risultata pari al 52.9% relativamente alla totalità dei bambini sordi esaminati (9/17); di questi 4 erano senza causa di sordità o fattori di rischio noti, 4 presentavano sintomatologia severa per CMV (SGA, calcificazioni cerebrali, dilatazione ventricolare, ipotonia) e 1 sintomatologia misconosciuta per CMV.
Accurate determination of hepatitis C virus RNA level is essential for evaluating the response to... more Accurate determination of hepatitis C virus RNA level is essential for evaluating the response to antiviral therapy, to determine the duration of treatment, and to predict treatment outcome. Currently, two real-time based polymerase chain reaction assays are used widely to monitor the hepatitis C RNA level: the Abbott RealTime HCV assay and the Cobas Taqman HCV assay. Recently, a third assay has become commercially available: the Artus HCV QS-RGQ assay, which uses the QIAsymphony SP/AS platform for sample preparation and PCR-setup, and the Rotor-Gene Q for amplification and detection. In this study, the performance of the Artus HCV QS-RGQ assay was tested on 105 plasma samples and compared to that of the Cobas Taqman HCV assay. Linear regression analysis showed a good agreement between the two assays. A slightly better sensitivity was observed with the Cobas Taqman assay, while higher hepatitis C viral RNA levels were measured by the Artus HCV QS-RGQ assay in samples positive for hepatitis C genotypes 4. Taken together, the data suggest that the Artus HCV QS-RGQ assay is useful in a diagnostic setting. The combination with the versatile QIAsymphony SP/AS system may represent a major advantage for clinical virological laboratories aiming at optimizing their workflow.
HIV-1 viral load determination is a crucial step for monitoring the efficacy of highly active ant... more HIV-1 viral load determination is a crucial step for monitoring the efficacy of highly active antiretroviral therapy (HAART) and predicts disease progression. Real-time PCR based assays are available for monitoring the viral load. They differ in sensitivity, genomic target region and dynamic range. In this study, the performance of the Roche Cobas Taqman HIV-1 v2.0 was evaluated on plasma samples from HIV-1 positive patients in parallel with the Abbott RealTime HIV-1 assay in a routine diagnostic setting. Overall, there was a good agreement between the two assays. However, some samples detected by the Abbott RealTime HIV-1 assay but below the limit of quantitation of the assay were found negative result when tested with the Roche Cobas Taqman HIV-1 v2.0. It is conceivable that signal anomalies or background noise may affect the lower-end precision of the Abbott RealTime HIV-1 assay. Based on these results, it is concluded that it is not recommended to switch platform during longitudinal viral load monitoring of HIV-1 positive patients.
International Journal of Immunopathology and Pharmacology, Oct 1, 2012
Human papillomavirus (HPV) is the most common sexually transmitted agent worldwide. Prevalence va... more Human papillomavirus (HPV) is the most common sexually transmitted agent worldwide. Prevalence varies according to the geographic regions, and is highest in developing countries. Geographic differences exist also in the detection rate of oncogenic types in malignant cervical lesions. In this study, the prevalence of HPV infection as well as the spectrum of HPV types was evaluated in Italian and immigrant women of the urban area of Rome. Several risk factors (age at first intercourse, number of partners, smoking, pregnancy, age at first pregnancy, contraception, education, and menarche) were taken into consideration. Overall, there was a high prevalence of HPV infection in the two groups studied. No significant differences were observed in the spectrum ofHPV types detected. HPV 16 and 18 were the types detected more frequently in both groups. Interestingly, HPV 54 and 70 were found only in the immigrants. Whether this finding reflects a recent introduction of these HPV types in the population studied remains to be established. Monitoring of HPV types in the population is advisable, especially in countries like Italy which is a destination and a gateway for immigrants directed towards north and central Europe. The introduction of high risk HPV variants may have a clinical impact and affect the diagnostic procedures.
Objective: Oncogenic human papillomaviruses (HPVs) are the etiological agents of cervical cancer.... more Objective: Oncogenic human papillomaviruses (HPVs) are the etiological agents of cervical cancer. Different cofactors might be needed for malignant transformation, but they still remain elusive. Methods: To delineate the role of Chlamydia trachomatis (CT) and herpes simplex virus type 2 (HSV2) in HPV-positive cervical intraepithelial neoplasia (CIN) lesions and cervical carcinoma a series of 149 cervical cancer and CIN biopsies were analyzed for CT and HSV2 DNA by PCR, and HPV genotyped by InnoLipa. Monitoring of aberrations in key intracellular pathways due to CT/HSV2 and HPV co-expression were analyzed with 13 biomarkers. Results: Of the 149 samples tested, 136 were HPV DNA positive; 32/136 contained also CT DNA and 29 HSV2 DNA. Detection of CT was significantly (p = 0.0001) related to multiple-type HPV infections, while HSV2 was of borderline significance (p = 0.053). Of the 13 biomarkers tested, cytoplasmic and nuclear NF-ĸB and VEGF-C were significantly increased in CT+/HPV+ lesions; p = 0.023, p = 0.045, and p = 0.020 as well as survivin, p = 0.026. Survivin was the only marker that was overexpressed also in HSV2+/HPV+ lesions, p = 0.027. Conclusions: CT infection favors the entry and persistence of multiple HR-HPV types, which leads to viral integration, inhibition of apoptosis, overexpression of E6/E7 oncogenes and cell transformation.
Background: In the last two years, the SARS-CoV-2 pandemic has determined radical changes in huma... more Background: In the last two years, the SARS-CoV-2 pandemic has determined radical changes in human behaviors and lifestyles, with a drastic reduction in socialization due to physical distancing and self-isolation. These changes have also been reflected in the epidemiological patterns of common respiratory viruses. For this reason, early discrimination of respiratory viruses is important as new variants emerge. Methods: Nasopharyngeal swabs of 2554 patients, with clinically suspected Acute Respiratory Infections (ARIs) from October 2019 to November 2021, were collected to detect 1 or more of the 23 common respiratory pathogens, especially viruses, via BioFilmArray RP2.1plus, including SARS-CoV-2. Demographical characteristics and epidemiological analyses were performed as well as a laboratory features profile of positive patients. Results: An observational study on 2300 patients (254 patients were excluded because of missing data) including 1560 men and 760 women, median age of 64.5 ...
E-cadherin plays a pivotal role in maintenance of normal adhesion in epithelial cells but has als... more E-cadherin plays a pivotal role in maintenance of normal adhesion in epithelial cells but has also been shown to suppress tumour invasion and participate in cell signalling. Known to be capable of reversing the invasive phenotype of high-risk human papillomavirus (HPV)-transformed keratinocytes, E-cadherin is down-regulated in CIN and cervical cancer (CC), but still incompletely studied as an intermediate endpoint marker in this disease. As part of our HPV-PathogenISS study, a series of 150 CCs and 152 CIN lesions were examined using immunohistochemical (IHC) staining for E-cadherin, and tested for HPV using PCR with three primer sets (MY09/11, GP5+/GP6, SPF). Follow-up data were available from all squamous cell carcinoma (SCC) patients, and 67 CIN lesions were monitored with serial PCR for HPV after cone treatment. Expression of E-cadherin was reduced in parallel with the increasing grade of CIN, with major down-regulation upon transition to CIN3 and further to invasive cancer (OR ...
In women with HIV-associated immunosuppression, HPV infections have an increased risk of progress... more In women with HIV-associated immunosuppression, HPV infections have an increased risk of progression to high-grade cervical intraepithelial neoplasia (CIN). With the HAART-induced prolonged survival and more protracted clinical course of AIDS, progression of CIN to cervical cancer (CC) has become a clinically relevant issue, and the mechanisms responsible for HIV-HPV interactions need further elucidation. The study design and analysis of the baseline data of our new project are presented. This project is a combination of a prospective cohort study of HIV- and HIV+ women, and a retrospective analysis of CIN lesions and cervical cancer. Up to the present, 244 women have been enrolled (17 HIV+) and subjected to epidemiological interview, colposcopic examination, sampling for HPV testing and typing (PCR, InnoLiPA), and HPV serology. The retrospective series of biopsies were analysed for 13 biomarkers (monitoring key molecular events) using immunohistochemistry and tested for HPV by PCR ...
Several detailed algorithms for the appropriate use of human papillomavirus (HPV) testing in the ... more Several detailed algorithms for the appropriate use of human papillomavirus (HPV) testing in the management of women with abnormal Pap (Papanicolaou) smears have been launched, but their direct country-to-country adoption is difficult. This necessitates their testing in individual settings, which is ongoing in our colposcopy referral clinic. A series of 224 consecutive women attending the clinic with the usual referral indications (ASC-US or higher in Pap) were examined by the conventional diagnostic tools (PAP smear, colposcopy, punch biopsy) and subjected to HPV testing and viral typing for both low-risk (L-R) and high-risk (H-R) types by nested PCR-based techniques. Predictors of the high-grade diagnostic categories were analysed using both univariate- and multivariate modelling, and the performance characteristics (sensitivity, specificity, NPV, PPV) of all tests in detecting high-grade CIN were calculated. In the PAP test, ASC-US smears were most common (37.9%), followed by low...
In recent years, efforts have been made to identify molecular markers as potential screening tool... more In recent years, efforts have been made to identify molecular markers as potential screening tools in the early detection of cervical cancer precursors. One-hundred-eighty-two women admitted to the Colposcopy Unit of Tor Vergata University Hospital were enrolled in this study. The inclusion criteria were: i) Pap test with atypical squamous cells of undetermined significance (ASCUS) or low-grade squamous intraepithelial lesion (LSIL); ii) normal cytology but human papillomavirus (HPV) DNA test positive for at least one of the most frequently detected five high-risk HPV types (16, 18, 31, 33 and 45). HPV DNA was detected with the HPV Sign kit and the type was assigned by pyrosequencing using the PyroMark ID System. E6/E7 transcripts of the high-risk HPV types 16, 18, 31, 33 and 45 were detected by the NucliSense EasyQ HPV kit. Overall, 90 (49.5%) patients were HPV-DNA negative, whereas 92 (50.5%) were HPV-DNA positive. Single infections were detected in 55 women: HPV 16 ranked first (...
Novel respiratory viruses have been identified as possible agents of upper and lower respiratory ... more Novel respiratory viruses have been identified as possible agents of upper and lower respiratory tract infections. Multiplex real-time PCRs have been developed to identify clinically relevant respiratory pathogens. In this study, 178 respiratory samples already screened for influenza virus types A and B by Flu A/B ASR real-time PCR kit were retrospectively analyzed with the Respiratory Multi Well System (MWS) r-gene™ real-time PCR kit which detects a wide spectrum of respiratory pathogens. The goal was to demonstrate the importance of a wide spectrum screening compared to a single diagnostic request. The Flu A/B ASR kit detected influenza B virus in 1.7% of the samples (3/178) and no influenza A virus. The MWS r-gene™ kit detected influenza virus in 6.7% (12/178) of samples (0.6% influenza A, and 6.2% influenza B), while the overall detection rate for respiratory pathogens was 54% (96/178). Co-infections were detected in 8/178 (4.5%) samples. Adenovirus was the infectious agent dete...
Anyplex™II HPV28 is a new PCR assay designed for HPV genotyping. It can detect 28 HPV types inclu... more Anyplex™II HPV28 is a new PCR assay designed for HPV genotyping. It can detect 28 HPV types including 19 high-risk and 9 low-risk types. This study evaluated the performance of Anyplex™II HPV28 on 123 fresh cervical samples screened in parallel with HPV Sign® Genotyping Test. Of the 123 samples screened, 93 were positive, 15 negative, and 15 discordant. The total number of HPV positive samples combined was 108: 38 single infections and 70 multiple infections. The agreement between the two tests was 87.8%, κ=0.592. Genotype specific agreement was strong for HPV 16 (k=0.761), HPV 18 (k=0.674), and HPV 35 (k=0.796). Sensitivity and specificity of Anyplex™II HPV28 assay using HPV Sign® Genotyping Test as reference was 84.8% and 94%; conversely, sensitivity and specificity of HPV Sign® Genotyping Test was 29% and 99.5%. Anyplex™II HPV28 assay is a sensitive and specific assay suitable for HPV genotyping but requires clinical validation.
Uploads
Papers by Pierpaolo Paba