The use of a specific DNA probe and polymerase chain reaction for the detection of Mycobacterium leprae.
A DNA probe encoding approximately 80% of the 18-kDa protein gene of Mycobacterium leprae was isolated and tested for specificity by assessing hybridization of the probe to genomic DNA from taxonomically related and unrelated DNA samples. The 360-base-pair (bp) probe was specific for M. leprae DNA and did not hybridize with genomic DNA from 18 species of bacteria nor with DNA from human, murine, and armadillo sources. Oligonucleotide primers were synthesized corresponding to the 5' and 3' ends of the 360-bp fragment to yield a fragment of similar size on amplification of M. leprae DNA by the polymerase chain reaction (PCR). A simple procedure for DNA extraction from M. leprae-infected tissues was developed that provided suitable template DNA for amplification. The PCR test was specific for M. leprae DNA from human and murine sources and detected M. leprae DNA in biopsies from leprosy patients and from control and uninfected human skin biopsy preparations seeded with as few as 100 M. leprae.
Laboratory Research Branch, Gillis W. Long Hansen's Disease Center, Carville, LA 70721.
D L
DL
Williams
T P
TP
Gillis
R J
RJ
Booth
D
D
Looker
J D
JD
Watson
D000818:Animals; D001345:Autoradiography; D001483:Base Sequence; D015342:DNA Probes; D004269:DNA, Bacterial / Q000032:analysis*; D004587:Electrophoresis, Agar Gel; D005784:Gene Amplification; D006801:Humans; D007918:Leprosy / Q000175:diagnosis*; D008969:Molecular Sequence Data; D009166:Mycobacterium leprae / Q000235:genetics / Q000302:isolation & purification*; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction; D011237:Predictive Value of Tests; D012867:Skin / Q000382:microbiology; D013045:Species Specificity
D015342:DNA Probes; D004269:DNA, Bacterial