Clinical utility of LSR/A15 gene for Mycobacterium leprae detection in leprosy tissues using the polymerase chain reaction.
Skin biopsy and slit-skin smears from 46 leprosy patients and 4 nonleprosy patients were tested for the presence of Mycobacterium leprae by the polymerase chain reaction (PCR) using primers based on the sequence of the LSR/15 kD gene. The PCR was found to be specific and sensitive, with a detection level of 10 and 100 bacilli. PCR using skin biopsies gave a higher detection rate than did slit-skin smears, probably due to the higher density of bacilli in a 4-mm punch biopsy. Dot blot hybridization with radioactive probes was 10-fold more sensitive than the ethidium bromide staining. Eight patients who did not show acid-fast bacilli in tissues by the conventional methods were shown to have PCR-amplified M. leprae DNA. False-negative results were obtained in 3 cases even though formal evidence for tissue inhibitors was absent.
Misra
N
N
Department of Biotechnology, All India Institute of Medical Sciences, New Delhi.
Ramesh
V
V
Misra
R S
RS
Narayan
N P
NP
Colston
M J
MJ
Nath
I
I
D001483:Base Sequence; D015342:DNA Probes / Q000737:chemistry; D004269:DNA, Bacterial / Q000032:analysis*; D004587:Electrophoresis, Agar Gel; D005188:False Negative Reactions; D005798:Genes, Bacterial; D006801:Humans; D007917:Leprostatic Agents / Q000627:therapeutic use; D007918:Leprosy / Q000175:diagnosis* / Q000188:drug therapy / Q000382:microbiology; D008969:Molecular Sequence Data; D009166:Mycobacterium leprae / Q000235:genetics* / Q000302:isolation & purification; D016133:Polymerase Chain Reaction; D011237:Predictive Value of Tests; D012680:Sensitivity and Specificity; D012867:Skin / Q000382:microbiology*
D015342:DNA Probes; D004269:DNA, Bacterial; D007917:Leprostatic Agents