C - Chemistry – Metallurgy – 12 – Q
Patent
C - Chemistry, Metallurgy
12
Q
C12Q 1/68 (2006.01)
Patent
CA 2354234
Amplification and cycle sequencing primer sets have been developed for the detection and analysis of antibiotic resistance-associated mutations in defined regions of the rpoB (rifampin), katG (isoniazid), oxyR-ahpC PR (isoniazid), mabA (isoniazid), rpsL/s12 (streptomycin), 16S/rrs (streptomycin), embB (ethambutol), pncA (pyrazinamide), gyrA (ciprofloxacin) and 23S (azithromycin) genes of Mycobacterium tuberculosis. These primers can be used in a method for detection and characterization of Mycobacterium tuberculosis present in a sample. The method includes the steps of obtaining a sputum sample suspected of containing M. tuberculosis, performing a first sequencing procedure, with or without prior amplification, on the sample to detect the presence of M. tuberculosis, and if present to evaluate the rpoB, katG, rpsL/s12 and 23S genes for the presence of antibiotic-resistance inducing mutations; and (c) if M. tuberculosis is detected in step (b), performing a second sequencing procedure, with or without prior amplification, on the sample to evaluate the additional genes for the presence of antibiotic- resistance inducing mutations.
Cette invention concerne des ensembles d'armoceurs de s~quen×age de cycle et d'amplification qui permettent de d~tecter et d'analyser des mutations associ~es ~ une r~sistance aux antibiotiques dans des r~gions pr~d~termin~es des gnes rpoB (rifampine), katG (isoniazide), oxyR-ahpC PR (isoniazide), mabA (isoniazide), rpsL/s12 (streptomycine), 16S/rrs (streptomycine), embB (~thambutol), pncA (pyrazinamide), gyrA (ciprofloxacine) et 23S (azithromycine) du Mycobacterium tuberculosis. Ces amorceurs peuvent Útre utilis~s dans un proc~d~ de d~tection et de caract~risation de Mycobacterium tuberculosis dans un ~chantillon. Ce proc~d~ consiste ~ obtenir un ~chantillon d'expectoration que l'on suspecte de contenir le M. tuberculosis, ~ effectuer une premire proc~dure de s~quen×age, avec ou sans amplification pr~alable, sur l'~chantillon afin de d~tecter la pr~sence de M. tuberculosis et, si ce dernier est pr~sent, ~ ~valuer les gnes rpoB, katG, rpsL/s12 et 23S afin de d~celer la pr~sence d'une r~sistance aux antibiotiques entra¹nant des mutations. Si (c) l'on d~tecte la pr~sence de M. tuberculosis au cours de l'~tape (b), on effectue une seconde proc~dure de s~quen×age, avec ou sans amplification pr~alable, sur l'~chantillon afin d'~valuer les gnes additionnels et de d~celer la pr~sence d'une r~sistance aux antibiotiques entra¹nant des mutations.
Bayer Healthcare Llc
Torys Llp
Visible Genetics Inc.
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