TY - JOUR KW - Animals KW - Autoradiography KW - Base Sequence KW - DNA Probes KW - DNA, Bacterial KW - Electrophoresis, Agar Gel KW - Gene Amplification KW - Humans KW - leprosy KW - Molecular Sequence Data KW - Mycobacterium leprae KW - Nucleic Acid Hybridization KW - polymerase chain reaction KW - Predictive Value of Tests KW - Skin KW - Species Specificity AU - Williams D L AU - Gillis T P AU - Booth R J AU - Looker D AU - Watson J D AB -

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.

BT - The Journal of infectious diseases C1 - http://www.ncbi.nlm.nih.gov/pubmed/2192005?dopt=Abstract DA - 1990 Jul DO - 10.1093/infdis/162.1.193 IS - 1 J2 - J. Infect. Dis. LA - eng N2 -

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.

PY - 1990 SP - 193 EP - 200 T2 - The Journal of infectious diseases TI - The use of a specific DNA probe and polymerase chain reaction for the detection of Mycobacterium leprae. VL - 162 SN - 0022-1899 ER -