Extraction and detection of Mycobacterium leprae DNA from ZNCF-stained skin smear slides for better identification of negative skin smears.
Abstract Identification of Mycobacterium leprae, which causes leprosy, is done by Ziehl Neelsen Carbol Fuchsin (ZNCF) stained slit skin smear microscopy that aids in the diagnosis and quantification of approximate bacterial load carried by the patient. We attempted M. leprae DNA extraction from 46 stained slit skin smear negative slides, using Proteinase K and SDS lysis, followed by ethanol precipitation. M. leprae specific primers (16SrRNA) were used for PCR-based amplification of DNA. We could detect M. leprae DNA in 15 (32.6%) samples. The method can be useful in the diagnosis of apparently slit skin smear negative leprosy cases.
Kamble
R R
RR
Stanley Browne Laboratory, TLM Community Hospital, Nand Nagari Delhi - 110 093, India.
Shinde
V S
VS
Madhale
S P
SP
Kamble
A A
AA
Ravikumar
B P
BP
Jadhav
R S
RS
D017931:DNA Primers / Q000235:genetics; D004269:DNA, Bacterial / Q000302:isolation & purification*; D003902:Detergents / Q000494:pharmacology; D019286:Endopeptidase K / Q000378:metabolism; D006801:Humans; D007918:Leprosy / Q000175:diagnosis* / Q000382:microbiology; D009166:Mycobacterium leprae / Q000235:genetics*; D057089:Pathology, Molecular / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012329:RNA, Bacterial / Q000235:genetics; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D012867:Skin / Q000382:microbiology* / Q000473:pathology*; D012967:Sodium Dodecyl Sulfate / Q000494:pharmacology
D017931:DNA Primers; D004269:DNA, Bacterial; D003902:Detergents; D012329:RNA, Bacterial; D012336:RNA, Ribosomal, 16S; D012967:Sodium Dodecyl Sulfate; D019286:Endopeptidase K