Papers by Ravindra Turankar
Archives of Clinical Microbiology, Sep 18, 2017
Infection and Drug Resistance, Dec 1, 2018
Infection and Drug Resistance, 2018
Despite more than three decades of multidrug therapy (MDT), leprosy remains a major public health... more Despite more than three decades of multidrug therapy (MDT), leprosy remains a major public health issue in several endemic countries, including India. The emergence of drug resistance in Mycobacterium leprae (M. leprae) is a cause of concern and poses a threat to the leprosy-control program, which might ultimately dampen the achievement of the elimination program of the country. Rifampicin resistance in clinical strains of M. leprae are supposed to arise from harboring bacterial strains with mutations in the 81-bp rifampicin resistance determining region (RRDR) of the rpoB gene. However, complete dynamics of rifampicin resistance are not explained only by this mutation in leprosy strains. To understand the role of other compensatory mutations and transmission dynamics of drug-resistant leprosy, a genome-wide sequencing of 11 M. leprae strains-comprising five rifampicin-resistant strains, five sensitive strains, and one reference strain-was done in this study. We observed the presence of compensatory mutations in two rifampicin-resistant strains in rpoC and mmpL7 genes, along with rpoB, that may additionally be responsible for conferring resistance in those strains. Our findings support the role for compensatory mutation(s) in RNA polymerase gene(s), resulting in rifampicin resistance in relapsed leprosy patients.
Frontiers in tropical diseases, Aug 19, 2022
Introduction: Molecular epidemiology of leprosy is very important to study leprosy transmission d... more Introduction: Molecular epidemiology of leprosy is very important to study leprosy transmission dynamics and to enhance our understanding of leprosy in endemic areas by utilizing the molecular typing method. Nowadays our understanding of leprosy transmission dynamics has been refined by SNP typing and VNTR marker analysis of M. leprae strains. Objective: This study was carried out to find out the presence of viable M. leprae in the soil and water samples from residing areas of leprosy patients staying in different blocks of Purulia district of West Bengal, understanding their genotypes and compared with that of M. leprae present in patients. Material and methods: Slit-skin smear (SSS) samples (n=112) were collected from the active multibacillary leprosy patients from different blocks of leprosy endemic area. Soil samples (n=1060) and water samples (n=620) were collected from residing areas of leprosy patients. SNP subtyping was performed by PCR followed by sequencing. Multiplex PCR was performed using fifteen ML-VNTR loci and results were analysed. Results: We observed high PCR positivity in soil samples (344 out of 1060; 32%) and water samples (140 out of 620; 23%). These PCR positive samples when further screened for viability, it was observed that 150 soil samples (44%) and 56 water samples (40%) showed presence of 16S rRNA. SNP typing of M. leprae revealed presence of predominantly type 1. SNP subtype 1D (83%) was most prevalent in all the blocks of Purulia followed by subtype 1C (15%) and subtype 1A (2%). SNP subtype 2F was noted in only one sample. SNP and VNTR combination showed presence of similar strain type in certain pockets of Purulia region which was responsible for transmission.
Frontiers in Tropical Diseases, 2023
The collection of papers in this Research Topic provides insight
into the challenges associated ... more The collection of papers in this Research Topic provides insight
into the challenges associated with the diagnosis, prevention, and
management of mycobacterial diseases. The importance of recent
molecular techniques used for the rapid diagnosis and identification
of specific species and the transmission aspect of mycobacterial
disease is highlighted, from the individual to the community level.
The completion of treatment is essential to stop the transmission of
the disease and to facilitate its proper management. The
advancement in molecular technology and recognition of the
molecular marker in the M. leprae genome has added to the value
of tracking community transmission.
Indian Journal of Dermatology, Venereology and Leprology, Sep 8, 2022
Journal of Global Antimicrobial Resistance
Indian Journal of Medical Microbiology, 2016
to official reports received during 2012 from 105 countries and territories, the globally registe... more to official reports received during 2012 from 105 countries and territories, the globally registered prevalence of leprosy at the beginning of 2012 stood at 181,941 cases whereas the number of new cases detected during 2011 was 219,075. [2] However, some pockets of endemicity do remain in certain areas of Angola,
European Journal of Clinical Microbiology & Infectious Diseases
Journal of Global Antimicrobial Resistance, 2020
Journal of Global Antimicrobial Resistance, 2020
Diagnostic Microbiology and Infectious Disease, 2019
Early diagnosis of leprosy is important for limiting the severity of disease, which may lead to d... more Early diagnosis of leprosy is important for limiting the severity of disease, which may lead to disabilities and deformities if not treated timely. Multiplex PCR employing more than one gene, specific to target DNA, is more efficient detection tool. In the present study, slit skin scrapings, blood, nasal swabs and saliva from Paucibacillary (PB) and Multibacillary (MB) cases as well as household contacts of PB cases were tested by multiplex PCR using three different gene targets namely RLEP, 16SrRNA and sodA. We found an increase in overall diagnostic positivity for M. leprae DNA detection by M-PCR as compared to individual PCR. In case of nasal swabs using M-PCR the PPV, NPV were 0.5454, 0.8333 respectively. There is remarkable increase in PPV in SSS of PB cases and nasal swabs of HHCs using M-PCR. Conclusively, our finding suggests the utility of M-PCR for early diagnosis and household contact surveillance for leprosy.
Journal of global antimicrobial resistance, Jan 30, 2017
Emergence of multi drug resistant (MDR) organism for any infectious disease is a public health co... more Emergence of multi drug resistant (MDR) organism for any infectious disease is a public health concern. Global efforts to control leprosy by intensive chemotherapy have led to a significant decrease in the number of registered patients. Currently recommended control measures for treating leprosy with MDT was designed to prevent the spread of dapsone resistant strains of Mycobacterium leprae. We report here the identification of MDR strains of M. leprae from relapsed leprosy patients from endemic regions in India. The drug resistant profiles of the isolated strains were confirmed by the identification of mutations in genes previously shown to be associated with resistance to each drug (Rifampicin, Dapsone and Ofloxacin). Between 2009 and 2016, slit- skin smears samples were collected from 239 relapse and 11 new leprosy cases from hospitals of The Leprosy Mission across India. DNAs were extracted from these samples and were analyzed for PCR targeting genes rpoB, folP and gyrA associat...
Aim: Emergence of drug resistant Mycobacterium leprae (M.leprae) poses a threat to the treatment ... more Aim: Emergence of drug resistant Mycobacterium leprae (M.leprae) poses a threat to the treatment and control strategies for combating leprosy. Presence of point mutations within the drug resistance determining regions of M. leprae genome confer molecular basis of drug resistance to Dapsone, Rifampicin & Ofloxacin which are further confirmed with convectional mouse foot pad methods. This study focused on the identification of mutations within the rpoB gene region that are specific for rifampicin interaction and further in-silico analysis was carried out to determine the variations in the interactions. Materials and Methods: DNA and RNA were isolated from slit skin scrapings of 60 relapsed leprosy patients from The Leprosy Hospitals (TLM) in India. PCR targeting rpoB gene region and amplicon sequencing was performed to determine point mutations. mRNA expression levels of rpoB and High Resolution Melt Analysis of mutants was performed using Rotor Gene Q Realtime PCR. In-silico docking ...
Infection, Genetics and Evolution, 2019
Background Mycobacterium leprae being an obligate intracellular parasite cannot be cultured in an... more Background Mycobacterium leprae being an obligate intracellular parasite cannot be cultured in any artificial culture media but it has been shown to reside in wild armadillos in North America. Many studies suggested that M. leprae could be found in the environment and may have a role in continuing transmission of the disease. The exact role of the environment in the transmission dynamics is still speculative. The present study was undertaken to find out the presence of viable M. leprae around patients' environment like soil and water and association of free living pathogenic protozoa, Acanthamoeba which might play an important role in transmission of the disease. Methods Seven hundred soil and 400 water samples were collected from the surroundings of the houses of leprosy patients from endemic villages. Two hundred soil and 80 water samples were also collected from the surroundings of normal inhabitants from non-endemic villages as controls. These samples were screened for the presence of M. leprae and Acanthamoeba using DNA PCR detection of M. leprae and Acanthamoeba.
PLOS Neglected Tropical Diseases, 2018
Leprosy is a chronic infectious disease caused by Mycobacterium leprae and mainly affects skin, p... more Leprosy is a chronic infectious disease caused by Mycobacterium leprae and mainly affects skin, peripheral nerves. Vitamin D receptor (VDR) gene polymorphism has been found to be associated with leprosy. Vitamin D has been shown to control several host immunomodulating properties through VDR gene. Vitamin D deficiency was also found to be linked to an increased risk for several infections and metabolic diseases. In the present study, we investigated the association of VDR gene polymorphism, mRNA gene expression of VDR and the vitamin D levels with leprosy and its reactional states. A total of 305 leprosy patients consisting of tuberculoid (TT), borderline tuberculoid (BT), borderline lepromatous (BL), lepromatous leprosy (LL), as well as 200 healthy controls were enrolled in the study. We identified single nucleotide polymorphisms (SNPs) of VDR Taq1, Fok1 and Apa1, as well as the expression of VDR mRNA gene using PCR-based restriction fragment length polymorphism (RFLP) analysis and real-time PCR respectively. We also performed ELISA to measure vitamin D levels. We observed that SNP of VDR gene (Fok1 and Taq1) are associated with the leprosy disease. The allelic frequency distribution of T and t allele (p = 0.0037), F and f allele (p = 0.0024) was significantly higher in leprosy patients and healthy controls. ff genotype of Fok1 was found to be associated with leprosy patients [p = 0.0004; OR (95% CI) 3.148 (1.662-5.965)]. The recessive model of Fok1 genotype was also found to be significantly associated in leprosy patients in comparison to healthy controls [p = 0.00004; OR (95% CI) 2. 85 (1.56-5.22)]. Leprosy patients are significantly associated with t-F-a haplotype. Further, VDR gene expression was
Infection and drug resistance, 2018
Despite more than three decades of multidrug therapy (MDT), leprosy remains a major public health... more Despite more than three decades of multidrug therapy (MDT), leprosy remains a major public health issue in several endemic countries, including India. The emergence of drug resistance inis a cause of concern and poses a threat to the leprosy-control program, which might ultimately dampen the achievement of the elimination program of the country. Rifampicin resistance in clinical strains ofare supposed to arise from harboring bacterial strains with mutations in the 81-bp rifampicin resistance determining region (RRDR) of thegene. However, complete dynamics of rifampicin resistance are not explained only by this mutation in leprosy strains. To understand the role of other compensatory mutations and transmission dynamics of drug-resistant leprosy, a genome-wide sequencing of 11strains - comprising five rifampicin-resistant strains, five sensitive strains, and one reference strain - was done in this study. We observed the presence of compensatory mutations in two rifampicin-resistant st...
Infection, Genetics and Evolution, 2018
Non-tuberculous mycobacteria (NTM) are environmental mycobacteria found ubiquitously in nature. T... more Non-tuberculous mycobacteria (NTM) are environmental mycobacteria found ubiquitously in nature. The present study was conducted to find out the presence of various species of NTM in leprosy endemic region along with Mycobacterium (M) leprae. Water and wet soil samples from the periphery of ponds used by the community were collected from districts of Purulia of West Bengal and Champa of Chhattisgarh, India. Samples were processed and decontaminated followed by culturing on Lowenstein Jensen (LJ) media. Polymerase chain reaction (PCR) was performed using 16S rRNA gene target of mycobacteria and species was confirmed by sequencing method. Indirect immune-fluorescent staining of M. leprae from soil was performed using M. leprae-PGL-1 rabbit polyclonal antibody. The phylogenetic tree was constructed by using MAFFT software. From 380 soil samples 86 NTM were isolated, out of which 34(40%) isolates were rapid growing mycobacteria (RGM) and 52(60%) isolates were slow growing mycobacteria (SGM). Seventy-seven NTM isolates were obtained from 250 water samples, out of which 35(45%) were RGM and 42(55%) were SGM. Amongst all the RGM, we isolated M. porcinum, M. psychrotolerans, M. alsenase, M. arabinose and M. asiaticum from Indian environmental samples. M. fortuitum was the most commonly isolated species of all RGM. Out of all SGM, M. holsaticum, M. yongonense, M. seouense, M. szulgai, M. europaeum, M. simiae and M. chimaera were isolated for the first time from Indian environment. M. intracellulare was the commonest of all isolated SGM. Presence of M. leprae was confirmed by indirect immunofluorescent microcopy and PCR method from the same environmental samples. Phylogenetic tree was showing a close association between these NTMs and M. leprae in these samples. Several species of pathogenic and nonpathogenic NTM species along with M. leprae were isolated from soil and pond water samples from leprosy endemic regions and these might be playing a role in causing disease and maintaining leprosy endemicity in India.
International Journal of Mycobacteriology, 2019
Background: The aim of the present study was to isolate and characterize nontuberculous mycobacte... more Background: The aim of the present study was to isolate and characterize nontuberculous mycobacteria (NTM) on Lowenstein–Jensen media supplemented with glycerol or pyruvate on two different temperatures from soil samples from leprosy endemic tribal areas of Purulia. Methods: Mycobacterium leprae DNA was isolated from these samples followed by polymerase chain reaction (PCR) amplification using RLEP gene target specific to M. leprae. DNA was extracted from NTM cultures by lysis method. The presence of Mycobacterial DNA was confirmed by PCR using universal mycobacterial primer as 16S rRNA. NCBI nBlast was used for the authentication of NTMs, and phylogenetic tree was constructed using M. leprae and NTM species. Statistical Analysis Used: The percentile method and phylogenetic tree were used as stastical tool in this research article. Results: The rapid-growing mycobacteria (RGM) species, 4 (80%) was obtained more than that of slow growing mycobacteria (SGM) 1 (20%) supplemented on gly...
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Papers by Ravindra Turankar
into the challenges associated with the diagnosis, prevention, and
management of mycobacterial diseases. The importance of recent
molecular techniques used for the rapid diagnosis and identification
of specific species and the transmission aspect of mycobacterial
disease is highlighted, from the individual to the community level.
The completion of treatment is essential to stop the transmission of
the disease and to facilitate its proper management. The
advancement in molecular technology and recognition of the
molecular marker in the M. leprae genome has added to the value
of tracking community transmission.
into the challenges associated with the diagnosis, prevention, and
management of mycobacterial diseases. The importance of recent
molecular techniques used for the rapid diagnosis and identification
of specific species and the transmission aspect of mycobacterial
disease is highlighted, from the individual to the community level.
The completion of treatment is essential to stop the transmission of
the disease and to facilitate its proper management. The
advancement in molecular technology and recognition of the
molecular marker in the M. leprae genome has added to the value
of tracking community transmission.