Evaluation of polymerase chain reaction amplification of Mycobacterium leprae-specific repetitive sequence in biopsy specimens from leprosy patients.
Biopsy specimens were obtained from 102 leprosy patients before chemotherapy and examined by polymerase chain reaction (PCR) using the primers amplifying the 372-bp DNA of a repetitive sequence of Mycobacterium leprae. The PCR results were then compared with bacterial indices (BI) of slit-skin smears and biopsy specimens. The intensities of DNA bands were in general correlated with the numbers of acid-fast bacilli, and even a sample with only one organism gave a PCR positive result. Ten 5-micron sections from each frozen tissue sample were pooled and processed for DNA preparation. PCR was positive for 11 (73.3%) of 15 biopsy specimens with BI of 0 determined for the paraffin sections from the same biopsy samples. PCR also gave positive results for 84 (96.6%) of 87 BI positive biopsy samples. Although the difference in overall results between the two methods was not statistically significant, PCR seemed to have an advantage over microscopic examination in detecting M. leprae in biopsy specimens negative for acid-fast bacilli. Further evaluation of PCR using more specimens from leprosy patients who are bacteriologically negative is warranted to ensure PCR's advantage over the conventional microscopic examination for the diagnosis of leprosy.
Yoon
K H
KH
Department of Microbiology, Yonsei University College of Medicine, Seoul, Korea.
Cho
S N
SN
Lee
M K
MK
Abalos
R M
RM
Cellona
R V
RV
Fajardo
T T
TT
Guido
L S
LS
Dela Cruz
E C
EC
Walsh
G P
GP
Kim
J D
JD
D001483:Base Sequence; D001706:Biopsy; D004269:DNA, Bacterial / Q000032:analysis*; D005069:Evaluation Studies as Topic; D006801:Humans; D007918:Leprosy / Q000175:diagnosis* / Q000382:microbiology; D008969:Molecular Sequence Data; D009166:Mycobacterium leprae / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction; D012091:Repetitive Sequences, Nucleic Acid; D012680:Sensitivity and Specificity
D004269:DNA, Bacterial