Application of RLEP real-time PCR for detection of M. leprae DNA in paraffin-embedded skin biopsy specimens for diagnosis of paucibacillary leprosy.
The TaqMan real-time polymerase chain reaction (PCR) assay was evaluated systematically with respect to the standard curve, linear range, and used for detecting Mycobacterium leprae DNA in paraffin-embedded skin biopsy specimens from 60 confirmed leprosy patients and three healthy individuals and 29 other dermatoses and bacterial DNA from 21 different species. The test was further evaluated with 51 paucibacillary (PB) patients. The results showed that the test had good sensitivity (8 fg) and good specificity with no cross-reactivity with 21 other bacterial species and the control specimens, except one with Xanthomatosis. The real-time PCR detection rate for the 51 PB specimens was 74.5% (38 of 51). We conclude that the real-time PCR test is a useful adjunct test for diagnosing early stage or PB leprosy cases.
Capital University of Medicine Affiliated Beijing Friendship Hospital, Beijing Tropical Medicine Research Institute, Beijing, China; Yunnan Wenshan Skin Disease Dispensary, China; Department of Molecular Microbiology and Immunology, Bloomberg School of Public Health, Johns Hopkins University, Baltimore, Maryland.
Wen
W
Yan
Yan
Y
Xing
Lian Chao
LC
Yuan
Rong
R
De Yang
Fu Yue
FY
Tan
Ying
Y
Zhang
Huan-Ying
HY
Li
D000293:Adolescent; D000328:Adult; D000368:Aged; D058491:Bacterial Load; D001706:Biopsy; D016022:Case-Control Studies; D002648:Child; D004269:DNA, Bacterial / Q000032:analysis; D005260:Female; D006801:Humans; D020071:Interspersed Repetitive Sequences / Q000235:genetics; D056005:Leprosy, Paucibacillary / Q000175:diagnosis* / Q000382:microbiology; D008297:Male; D008875:Middle Aged; D009166:Mycobacterium leprae / Q000235:genetics*; D016612:Paraffin Embedding; D060888:Real-Time Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D012867:Skin / Q000382:microbiology*; D012871:Skin Diseases / Q000175:diagnosis / Q000382:microbiology; D055815:Young Adult
D004269:DNA, Bacterial