ABSTRACT
Background: In endemic regions of several countries, the prevalence of leprosy has not come down to the level of elimination. On the contrary, new cases are being detected in large numbers. Clinically, it is frequently noted that despite completion of multibacillary multidrug therapy for 12 months, the lesions remain active, especially in cases with high bacteriological indices. Aim: The present study focused on finding out the viable number of Mycobacterium leprae during the 12-month regimen of multibacillary multidrug therapy, at six and 12 months intervals and, attempting to determine their role in disease transmission. Methods: Seventy eight cases of multibacillary leprosy cases were recruited from leprosy patients registered at The Leprosy Mission hospitals at Shahdara (Delhi), Naini (Uttar Pradesh) and Champa (Chhattisgarh), respectively. Slit skin smears were collected from these patients which were transported to the laboratory for further processing. Ribonucleic acid was extracted by TRIzol method. Total Ribonucleic acid was used for real-time reverse transcription-polymerase chain reaction (two-step reactions). A standard sample with a known copy number was run along with unknown samples for a reverse transcription-polymerase chain reaction. Patients were further assessed for their clinical and molecular parameters during 6th month and 12th month of therapy. Results: All 78 new cases showed the presence of a viable load of bacilli at the time of recruitment, but we were able to follow up only on 36 of these patients for one year. Among these, using three different genes, 20/36 for esxA, 22/36 for hsp18 and 24/36 for 16S rRNA cases showed viability of M. leprae at the time of completion of 12 months of multidrug therapy treatment. All these positive patients were histopathologically active and had bacillary indexes ranging between 3+ and 4+. Patients with a high copy number of the Mycobacterium leprae gene, even after completion of treatment as per WHO recommended fixed-dose multidrug therapy, indicated the presence of live bacilli. Limitations: Follow up for one year was difficult, especially in Delhi because of the migratory nature of the population. Patients who defaulted for scheduled sampling were not included in the study. Conclusion: The presence of a viable load of bacilli even after completion of therapy may be one of the reasons for relapse and continued transmission of leprosy in the community
ABSTRACT
India attained the elimination figure of less than 1 case of leprosy per 10,000 people during December 2005. Despite this, India still accounts for the largest number of new leprosy cases in the world, maintaining more than 50 per cent of the leprosy burden of the world, notwithstanding over three decades of use of multidrug therapy. The present review analyzes the process of execution of the elimination program, identifies any lacunae therein and presents corrective measures that could be taken up for elimination of the disease from the country.
ABSTRACT
The symposium was organized by The Leprosy Mission Trust India (TLMTI) on the 29 November, 2017 at India Habitat Centre, Lodi Road, New Delhi, India. The meeting was organized for the stakeholders who are engaged in the leprosy elimination programme for an exposure to the recently emerging scenario on relapse and drug resistance in leprosy. A total of sixty three participants including representatives from Government of India, WHO, ILEP, GLRA, LEPRA India, NLR, DFIT, BLP, FMR, NIMHANS, ICMR-NJIL & OMD and ICMR gathered in the symposium. Most of the famous advisors of the country, leprologists and dermatologists of TLM, Safdarjung, Guru Tegh Bahadur hospitals, PGIMER (Chandigarh, India), DLO and SLO of Delhi participated in the symposium. Leprosy researchers and scientists from different organizations across the country also gathered for the deliberations and discussions. The symposium began with a welcome address from the Executive Director, TLM which was followed by inaugural keynote addresses by the experts of the country. Scientific deliberations by the leprologists, dermatologists and laboratory researchers were divided in four main sessions. This symposium had presentations on most current areas of importance such as goals and achievements of NLEP; diagnostics with focus on different forms of disease including neuritic leprosy; newer methods such as imaging for studying CNS involvement; response to therapy; drug resistance in the context of relapses, poor responders, reactions and multibacillary leprosy, transmission, methods and strategies for detection of drug resistance and its surveillance; national and global perspective etc. The Symposium concluded with a plenary session for the road map of a future strategy with recommendations which include molecular detection of drug resistance with focus on relapses, reactions and MB leprosy; confirmation of relevance of novel mutations in animals; networks for drug resistance surveillance and epidemiology of drug resistance in leprosy.
ABSTRACT
Background: The current strategy for leprosy control depends mainly on early case detection and providing the recommended multidrug therapy (MDT) dosage. Understanding the molecular mechanisms of drug resistance to each of these drugs is essential in providing effective treatment and preventing the spread of resistant strains in the community. The progress of molecular biology research provides a very effi cient opportunity for the diagnosis of drug resistance by in vitro method. Aim: We aimed to investigate the point mutations within the rpoB gene region of the Mycobacterium leprae genome, which are responsible for resistance to rifampicin, in order to determine the emergence of drug resistance in leprosy in the Kolkata region of West Bengal. Methods: A total of 50 patients with a relapse of leprosy were enrolled in the study. Skin smears were obtained for estimation of bacillary index and biopsies were obtained in 70% alcohol for extraction of DNA. The extracted DNA was amplifi ed by M. leprae-polymerase chain reaction (PCR) targeting rpoB gene region. Every single nucleotide base in the sequence is aligned to reference sequence and identity gaps were determined by NCBI – BLAST. Later in-silico analysis was done to identify the changes in the translated protein sequences. Results: A mutation at the base pair position 2275405 where G is replaced by C in the M. leprae genome, which corresponds to the coding region of rpoB gene (279 bp – 2275228 to2275506), was observed in two patients. This missense mutation in CAC codon brings about a glutamic acid to histidine change in the amino acid sequence of RNA polymerase beta subunit at the position 442 (Glu442His), a region specifi c for rifampicin interaction, which might be responsible for unresponsiveness to rifampicin by manifesting a stable bacteriological index in these 2 patients even after completion of 24 months of multibacillary multi-drug therapy (MB-MDT). Limitations: The major limitations of multipleprimer PCR amplifi cation refractory mutation system (MARS) assay is that it capable of detecting mutation at codon 425 and cannot distinguish any silent amino acid changes. Conclusion: The study indicates the existence of rifampicin drug resistance in Eastern India.