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Archives of Clinical Microbiology

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PCR amplification of genomic Mycobacterium leprae DNA by using different gene targets

Annual Conference on MICROBIAL PATHOGENESIS, INFECTIOUS DISEASE, ANTIMICROBIALS AND DRUG RESISTANCE
August 23-24, 2017 | Toronto, Canada

Vinay Kumar Pathak, Madhvi Ahuja, Ravindra Turankar, Itu Singh, Mallika Lavania and Utpal Sengupta

The Leprosy Mission Community Hospital, India

Posters & Accepted Abstracts: Arch Clin Microbiol

Abstract:

Several attempts have been made to establish a diagnostic test for leprosy since decades. None of these assays could diagnose more than 60% of PB cases or early cases of leprosy. The present study was attempted to develop a diagnostic test using M. leprae specific PCR in clinical samples. The study was aimed to detect M. leprae genomic DNA by using two different gene targets. Standardization for sensitivity of PCR for two genes was performed with standard genomic DNA of M. leprae strain NHDP-63. The standard DNA was serially diluted in 1:10 ratio up to 12 dilutions in decreasing concentrations. The DNA concentration of first dilution was 1x10-1 ?¼g/?¼l or 100 ng/?¼l to twelfth dilution 1x10-12 ?¼g/?¼l or 1 ?±g/?¼l. PCR amplification using two gene targets of M. leprae namely repetitive element rlep and 16S rRNA were performed with the same. PCR amplification for rlep gene was positive up to concentration of 1x10-9 ?¼g/?¼l or 1 fg/?¼l, similarly it was 1x10-10 ?¼g/?¼l or 100 ?±g/?¼l for 16S rRNA gene target. The sensitivity has been tested with clinical samples of leprosy patients and positivity of result was found 66.0% in case of rlep whereas it was 82.0% for 16S rRNA gene. In present study, PCR positivity for rlep and 16S rRNA gene were found efficient in the clinical samples and these gene targets can be further considered to develop a diagnostic tool for detection of sub clinical leprosy.

Biography :

Vinay Kumar Pathak has completed his MSc Biotechnology from Guru Nanak Dev University. Currently, he is pursuing his PhD at Stanley Browne Laboratory, The Leprosy Mission Community Hospital, Delhi. He is working as Senior Research fellow and has published one paper in a reputed journal.