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allied
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Ann Biol Sci, 2017
ISSN: 2348-1927
August 23-24, 2017 | Toronto, Canada
Annual Conference on
MICROBIAL PATHOGENESIS, INFECTIOUS DISEASE,
ANTIMICROBIALS AND DRUG RESISTANCE
S
everal 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
µg/µl or 100 ng/µl to twelfth dilution 1x10
-12
µg/µl
or 1 αg/µl. 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
µg/µl or 1 fg/µl,
similarly it was 1x10
-10
µg/µl or 100 αg/µl 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.
Speaker 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.
e:
pathakv.vp@gmail.comPCR amplification of genomic
Mycobacterium leprae
DNA by using different gene targets
Vinay Kumar Pathak, Madhvi Ahuja, Ravindra Turankar, Itu Singh, Mallika Lavania
and
Utpal Sengupta
The Leprosy Mission Community Hospital, India
Vinay Kumar Pathak et al., Arch Clin Microbiol, 8:5
DOI: 10.4172/1989-8436-C1-003