[PCR detection of Mycobacterium tuberculosis lacking IS 6110].
We have evaluated the frequency of M. tuberculosis strains which lack IS 6110 among 102 sputa isolated from Moroccan patients. A pair of primers was designed to amplify a 201bp DNA fragment of IS 6110. The amplified DNA was detected by ethidium bromide stained agarose gel electrophoresis and confirmed by southern blot hybridization with a 32P-labelled probe (PMTO2). To detect the presence of amplification inhibitors, an internal control DNA was added in each negative PCR result. Among 102 samples, 6 sputa were negative by PCR-IS 6110 but culture positive. The test of detection of M. tuberculosis for 2/6 sputa by PCR Amplicor amplifying 584 pb of rRNA 16s sequence was positive. RFLP analysis of these 2 strains revealed no bands hybridizing IS 6110 but PCR-Mt 308 was positive. These results confirmed that these M. tuberculosis strains are lacking IS 6110.
el Baghdadi
J
J
Unité des mycobactéries, Institut Pasteur du Maroc, Casablanca, Maroc.
Lazraq
R
R
Benani
A
A
Naciri
M
M
Ibrahimy
S
S
Benslimane
A
A
D000328:Adult; D015139:Blotting, Southern; D004396:Coloring Agents; D004269:DNA, Bacterial / Q000032:analysis*; D004587:Electrophoresis, Agar Gel; D004996:Ethidium; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D009018:Morocco; D009169:Mycobacterium tuberculosis / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction; D013183:Sputum / Q000382:microbiology*; D014397:Tuberculosis, Pulmonary / Q000175:diagnosis / Q000382:microbiology
D004396:Coloring Agents; D004269:DNA, Bacterial; D004996:Ethidium
Détection par PCR de Mycobacterium tuberculosis sans IS 6110.