Papers by Wijit Wonglumsom
Mycobacterium tuberculosis complex (MTBC) is a major causative agent of public health problems. N... more Mycobacterium tuberculosis complex (MTBC) is a major causative agent of public health problems. Nontuberculous mycobacteria are also increasingly encountered worldwide. Acid-fast bacilli (AFB) staining and culture are conventional methods used for mycobacterial identification. The aim of the study was to compare the performance of the two methods for AFB staining and culture, i.e. The hypertonic saline-sodium hydroxide (HS-SH) technique was compared with the N-acetyl-L-cysteine-sodium hydroxide (NALC-NaOH) method for AFB staining and cultivation. A cross-sectional study was conducted in a tuberculosis laboratory of the 3rd Office of Disease Prevention and Control, Nakhon Sawan Province, Thailand, during October 2015 to September 2016. Totally, 427 paired samples of sputum were digested and decontaminated by HS-SH and NALC-NaOH methods. After concentration, the processed samples were cultured in Lowenstein–Jensen (LJ) media and Mycobacteria Growth Indicator Tube (MGITs). The direct smear detection before and after concentration of the sputum by both methods was also examined. To evaluate HS-SH method, sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), accuracy and kappa were analyzed. The positive rates of AFB smears by both concentration methods were significantly higher than the direct method (P=0.032). The HS-SH method was highly comparable to the traditional NALC-NaOH method for microscopy and culture in LJ media and MGITs (kappa=0.96, 0.59 and 0.45, respectively) with no statistically significant difference. Sensitivity, specificity, PPV, NPV and accuracy of the HS-SH method for cultivation was 96.3%, 100.0%, 100.0%, 94.5% and 97.7%, respectively. The conclusion, The HS-SH method demonstrated good sensitivity, specificity, PPV, NPV and accuracy similar to NALC-NaOH method for sputum digestion and concentration of both microscopy and culture with low cost and workload.
Journal of the Medical Technologist Association of Thailand, 2018
Tuberculous meningitis (TBM) causes high mortality and serious complications due to delayed early... more Tuberculous meningitis (TBM) causes high mortality and serious complications due to delayed early diagnosis. This study evaluated a diagnostic value of adenosine deaminase activity (ADA) in cerebrospinal fluid (CSF) as a rapid screening test for TBM. Totally 121 patients with suspected symptoms of meningitis were included with 55 cases of TBM and 66 cases of non-TBM. With good performance verification of Diazyme enzymatic method used, ADA activity at a cut off value of 4.35 U/L was able to differentiate TBM from non-TBM by exhibiting a sensitivity of 89.1%, specificity of 80.3%, positive predictive value (PPV) of 79.0% and negative predictive value (NPV) of 89.8%. The mean ADA value was significantly higher ( p -value < 0.05) in TBM patients (9.65 ± 8.37 U/L; mean ± SD) than in non-TBM patients (4.08 ± 8.13 U/L; mean ± SD). The combination of ADA test and CSF examination increased the sensitivity to 96.3% and had a specificity of 63.6%. In conclusion, determination of cerebrospin...
Nowadays, clinical laboratory service has an important role in patient care management. A quality... more Nowadays, clinical laboratory service has an important role in patient care management. A quality test result must be obtained, but this can be affected by many factors. The correct process for specimen acquisition is very necessary and must be done by competent professional personnel. The objective of this study was to improve the competency of blood collection by phlebotomists through evidence-based learning. Evidence-based information gathering from nurses, medical technologists and clinical laboratory assistants was analyzed and synthesized to use as teaching material. Subjects were divided into two groups. The results showed that the trained groups of the clecturee type and the cevidence-basede type had different levels of improvement, with the more competent group being the evidence-based one. Five out of eight assessed parameters showed significant improvement. The overall score was increased from 49.79 to 73.08 percent In addition, the evidence-based group showed their impro...
Journal of the Medical Technologist Association of Thailand, 2016
Bacterial infection is a serious disease that may cause fatality in an infected patient. Bacteria... more Bacterial infection is a serious disease that may cause fatality in an infected patient. Bacterial culture is an important technique for diagnosis of infectious diseases. In bacterial culture process, errors can occur leading to incorrect reporting and affecting the patients directly. In this study, Failure Mode and Effects Analysis (FMEA) was used as a tool for systematic assessment and reduction of potential errors, risk management, and quality improvement of a process of bacterial cultivation in order to get accurate results. Failure modes that caused the incorrect reporting were analyzed and risk estimation calculated representing as risk priority number (RPN) for performing prioritization of the failure modes. In the culture process, 24 failure modes were issued and the RPN ranged from 5 to 500. The failure modes were categorized into 4 issues of pre-analytical phase, 7 issues of analytical phase and 4 issues of post-analytical phase. The top-seven RNP scores of failure modes i...
External quality assessment for evaluation of individual competency in isolation and identificati... more External quality assessment for evaluation of individual competency in isolation and identification of bacteria for clinical microbiological laboratory was studied. Lyophilized or freezedried bacteria were prepared and a panel of lyophilized bacteria was shipped once a year. The first set consisted of two mixed cultures and one pure culture. Forty-three laboratories participated in this first set. The second set provided three ampoules of lyophilized mixed bacteria. Thirty-eight laboratories participated in this second set. Fifteen (34.9%) laboratories and 21 (55.3%) laboratories were correctly reported for the first and second proficiency sets, respectively. Of 33 laboratories participated in both trials, fifteen (46%) laboratories showed performance improvement. A meeting concerning on quality assurance and good laboratory practices in clinical microbiology was also provided for participating laboratories before shipment of the second panel. This pilot scheme showed a high possibi...
International journal of food microbiology, Jan 2, 2018
The aim of this study was to develop a nanoparticle-based cell capture system combined with a lat... more The aim of this study was to develop a nanoparticle-based cell capture system combined with a lateral flow test strip (LFT) assay for rapid detection of Campylobacter jejuni from poultry samples. The developed assay was bench-marked against the standard modified Charcoal Cefoperazone Deoxycholate Agar (mCCDA) method according to ISO16140:2003 procedures. The synthesized ferromagnetic nanoparticles (FMNs) were modified with glutaraldehyde, then functionalized with polyclonal antibodies for specific C. jejuni capture and concentration from poultry samples. After lysing captured cells, DNA from C. jejuni was amplified by PCR using the primers designed to target the hipO gene, and the PCR amplicons were detected with the lateral flow test strip assay. Following the ISO16140:2003 guidelines, the relative detection limit, and the inclusivity and exclusivity tests were determined. The results showed that the limit of detection (LOD) of the assay was 10 or 1 cfu/ml with C. jejuni in pure cu...
Food Science and Biotechnology, 2016
Campylobacter is an important food-borne pathogen causing acute gastroenteritis worldwide. Magnet... more Campylobacter is an important food-borne pathogen causing acute gastroenteritis worldwide. Magnetic nanoparticle-based PCR coupled with streptavidin-horseradish peroxidase and a substrate was used for colorimetric detection. Forward primers conjugated to magnetic nanoparticles facilitated separation and concentration of Campylobacter DNA in a sample matrix. After PCR, a green color developed and was observed using the unaided eye, or detected using a spectrophotometer. High specificity and sensitivity of the 100 fg DNA/PCR reaction were achieved in pure culture experiments. The technique was applied for detection of Campylobacter on naturally contaminated chicken skin. All positive results were in agreement with results achieved using a conventional culture method. The magnetic nanoparticle-PCR-enzyme linked gene assay was practical and useful for detection of Campylobacter in complex matrices with PCR-interfering substances.
เวชสารแพทย์ทหารบก, 2012
Background : In order to obtaining a quality test result, specimen collection plays an important ... more Background : In order to obtaining a quality test result, specimen collection plays an important role. There are 2 professions generally involved in specimen collection; nurse and medical technologist. They are trained and practiced on their own system; eventually, gap among them might sometime lead to problem that has impacted to the patient. Objective : To solve problems on specimen collection, we decided to do the dual profession cotraining by the evidence-based learning method. Methods : A total of 12 sessions of 4 training process were offered continuously for 6 years. They were 1,231 participants; 861, 302, and 68 for nurses, medical technologists and others, respectively. An outcome of the training and their competency on specimen collection was assessed. Results : There was a great improvement on their competency after the co-training (p < 0.01). It was the best way on problem solving because it worked as pro-active process. In addition, there were transferred tacit knowledge between the two professions; a networking also settled to cooperate for future issues. As the viewpoint of trainers, knowledge and experiences learned from the co-training dual profession lead to several outcome; three textbooks and teaching materials related specimen collection had been written. There were at least 2 projects developed under the cooperation between the two professions and presented in a national meeting contest. Conclusion : The beneficial gains from dual professionals co-training were not only increased the trainee competency but also the trainer knowledge management.
Journal of the Medical Association of Thailand = Chotmaihet thangphaet, 2010
To become a quality clinical laboratory, personnel development is the most important factor. In o... more To become a quality clinical laboratory, personnel development is the most important factor. In order to achieve this goal, it should emphasize that clinical laboratory is not only a testing laboratory; it must be a knowledge-based service laboratory. A smart model for clinical laboratory personnel development under the Human Asset Development (HAD) program had been launched since 2003. To strengthen the competency of clinical laboratory personnel, an appropriate model was developed and apply to the clinical laboratory personnel. Medical technologist who currently worked in clinical laboratory participated in this study. The proposed model consisted of 3 phases. 1) The knowledge providing via update and refresher courses. 2) Application of learned knowledge to practice under close supervision. 3) Training on special topic and self oriented research activity. The outcome of 5 years project was evaluated. After the first phase, they were able to identify and solve their own troublesom...
The Southeast Asian journal of tropical medicine and public health, 2007
Escherichia coli was used to investigate quinolone resistance and mutations in gyrA gene of E. co... more Escherichia coli was used to investigate quinolone resistance and mutations in gyrA gene of E. coli isolated from pet (dog and cat), human (pet's owner), vegetable and edible ice in Bangkok and vicinity. Susceptibility test for nalidixic acid (NA) showed similar percent resistance among the sample sources but a lower ciprofloxacin (CIP) resistance was found particularly in human source. Mutations within quinolone resistance determining region of gyrA gene analyzed using non-radioactive single-strand conformation polymorphism (SSCP) and sequencing showed 10 different SSCP patterns. E. coli isolates from pet, vegetable and ice showed more variety of patterns than strains isolated from human. Four out of 10 SSCP patterns were identified as having mutations in amino acids positions 83 (Ser to Leu) and position 87 (Asp to Asn). These mutations were observed only in NA-resistant strains and combined mutations were observed only in E. coli isolated from humans and pets. As only 24% of ...
The Southeast Asian journal of tropical medicine and public health, 2007
A retrospective study of the patterns of antimicrobial susceptibility and phage types of 111 Salm... more A retrospective study of the patterns of antimicrobial susceptibility and phage types of 111 Salmonella typhi strains isolated in 1996 from Vietnam was carried out. The strains were tested for susceptibility to chloramphenicol, ampicillin, tetracycline, trimethoprim-sulfamethoxazole, nalidixic acid, ceftazidime, ceftriaxone and ciprofloxacin. Simultaneous resistance to chloramphenicol, ampicillin, tetracycline and trimethoprim-sulfamethoxazole were present in 84 strains (75.7%). Nalidixic acid resistance was only observed in 2 multidrug-resistant strains (1.8%). Twenty-one strains (18.9%) were completely susceptible to all drugs tested. All 111 strains were susceptible to ceftazidime, ceftriaxone and cipropfloxacin. The MIC values for chloramphenicol, ampicillin and trimethoprim-sulfamethoxazole corresponded with the results by disk diffusion method. On Vi phage-typing, 5 different phage types (28, A, D1, E1 and M1) were found in 12 strains (10.8%). However, most S. typhi strains we...
Biosensors and Bioelectronics, 2015
Pathogenic Vibrio cholerae produces a cholera toxin which is the cause of a severe diarrheal dise... more Pathogenic Vibrio cholerae produces a cholera toxin which is the cause of a severe diarrheal disease called &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;quot;Cholera&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;quot;. Available detection methods, including standard bacteriological test and immuno-based detection, are specific to the suspected pathogenic V. cholerae O1 and O139, but they are not specific to the cholera toxin producible strain. This work combined the polymerase chain reaction (PCR) of cholera toxin gene, ctxA gene, and microcantilever-based DNA sensor to improve the sensitivity and specificity of detection. Gold coated microcantilever, 250 µm long and 50 µm wide, with an embedded polysilicon wire acting as a piezoresistive material was modified by a self-assembled monolayer (SAM) of 3-mercaptopropionic acid (MPA) for immobilization of specific DNA probe via avidin layer on the surface. The avidin and 5&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;#39; biotinylated single-stranded DNA (ssDNA) probe concentrations were optimized for the immobilization at 50 µg/mL and 1 µM, respectively. The hybridization between ssDNA probe on this DNA sensor and target DNA creates nanomechanical bending and resistance change of piezoresistive material inside the beam. This microcantilever-based DNA sensor offers a detection sensitivity of 3.25 pg or 14 nM of DNA template for ctxA gene detection. The lowest number of V. cholerae O1 in food sample with and without the enrichment process that the polymerase chain reaction (PCR) for ctxA gene combined with this DNA sensor can detect is 0.835 and 835 cells/g, respectively. This detection sensitivity is 10 times higher than that of the conventional PCR method.
PLoS ONE, 2011
The ethanolic extract from Rhodomyrtus tomentosa leaf exhibited good antibacterial activities aga... more The ethanolic extract from Rhodomyrtus tomentosa leaf exhibited good antibacterial activities against both methicillinresistant Staphylococcus aureus (MRSA) and S. aureus ATCC 29213. Its minimal inhibitory concentration (MIC) values ranged from 31.25-62.5 mg/ml, and the minimal bactericidal concentration (MBC) was 250 mg/ml. Rhodomyrtone, an acylphloroglucinol derivative, was 62.5-125 times more potent at inhibiting the bacteria than the ethanolic extract, the MIC and MBC values were 0.5 mg/ml and 2 mg/ml, respectively. To provide insights into antibacterial mechanisms involved, the effects of rhodomyrtone on cellular protein expression of MRSA have been investigated using proteomic approaches. Proteome analyses revealed that rhodomyrtone at subinhibitory concentration (0.174 mg/ml) affected the expression of several major functional classes of whole cell proteins in MRSA. The identified proteins involve in cell wall biosynthesis and cell division, protein degradation, stress response and oxidative stress, cell surface antigen and virulence factor, and various metabolic pathways such as amino acid, carbohydrate, energy, lipid, and nucleotide metabolism. Transmission electron micrographs confirmed the effects of rhodomyrtone on morphological and ultrastructural alterations in the treated bacterial cells. Biological processes in cell wall biosynthesis and cell division were interrupted. Prominent changes including alterations in cell wall, abnormal septum formation, cellular disintegration, and cell lysis were observed. Unusual size and shape of staphylococcal cells were obviously noted in the treated MRSA. These pioneer findings on proteomic profiling and phenotypic features of rhodomyrtone-treated MRSA may resolve its antimicrobial mechanisms which could lead to the development of a new effective regimen for the treatment of MRSA infections.
Journal of Rapid Methods and Automation in Microbiology, 2001
Supplementation of Niroomand and Fung broth (NFB) and Hunt broth (HB) with Oxyrase @ , sheep bloo... more Supplementation of Niroomand and Fung broth (NFB) and Hunt broth (HB) with Oxyrase @ , sheep blood, hemin or yeast extract for recovery of Campylobacter was evaluated. Five strains of C. jejuni and one strain of C. coli were cultured in HB with or without Oxyrase ? NFB with or without Oxyrase @ , NFB without hemin, and NFB plus yeast extract without hemin. The HB without Oxyrase@ was evacuated and flushed three times with mixed gas (5% O,, 10% CO,, and 85% NJ prior to incubation. The effect of blood in HB supplemented with Oxyrase @ on the growth of Campylobacter was also examined. Hunt broth showed significantly better performance than NFB @ < 0.05). Supplementation of 0.2% hemin and 0.6% yeast extract stimulated growth by up to 2 log CFU/mL. depending on strains. Addition of 7% lysed blood enhanced growth of C. jejuni, and differences were significant at 6 h and 24 h of incubation. At 20 h of incubation, broth containing 0.6 units/ml of Oxyrase@ yielded Campylobacter counts of 8.6 to 10.9 log CFU/mL, and the broth flushed with mixed gas provided counts of 9.4 to 11.0 log CFU/mL. No statistical difference 0, > 0.05) was found between the Oxyrase @method and gas replacement method at incubation times of 12 h, 20 h, 28 h, and 36 h for all strains tested. Dissolved oxygen levels of all enrichment media were less than I. 5 rng/L. Using Oxyrase @supplemented HB without the cumbersome gassing step provides a simple and time-saving procedure for culturing Campylobacter jejuni and C. coli.
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Papers by Wijit Wonglumsom