US 2003/0152982 A1 · Method for the detection of the antibiotic resistance spectrum of Mycobacterium species

Abstract

Method for the detection of the antibiotic resistance spectrum of Mycobacterium species present in a sample, possibly coupled to the identification of the Micobacterium species involved, comprising the steps of: (i) if need be releasing, isolating or concentrating the polynucleic acids present in the sample; (ii) if need be amplifying the relevant part of the antibiotic resistance genes present in said sample with at least one suitable primer pair; (iii) hybridizing the polynucleic acids of step (i) or (ii) with at least one of the rpoB gene probes, as specified in table 2, under the appropriate hybridization and wash conditions; (iv) detecting the hybrids formed in step (iii); (v) inferring the Mycobacterium antibiotic resistance spectrum, and possibly the Mycobacterium species involved from the differential hybridization signal(s) obtained in step (iv).

Claims27

Dependent claims

Method for the detection of the antibiotic resistance spectrum of Mycobacterium species present in a sample, possibly coupled to the identification of the Mycobacterium species involved, comprising the steps of:

(i) if need be releasing, isolating or concentrating the polynucleic acids present in the sample;
(ii) if need be amplifying the relevant part of the antibiotic resistance genes present in said sample with at least one suitable primer pair as described in claims 22 to 24;
(iii) hybridizing the polynucleic acids of step (i) or (ii) with at least one of the rpoB gene probes, as specified in table 2, under appropiate hybridization and wash conditions;
(iv) detecting the hybrids formed in step (iii);
(v) inferring the Mycobacterium antibiotic resistance spectrum, and possibly the Mycobacterium species involved, from the differential hybridization signal(s) obtained in step (iv).

2 more elements

Method according to claim 1 for detection of rifampicin (and/or rifabutin) resistance of M. tuberculosis present in a biological sample, comprising the steps of:

(i) if need be releasing, isolating or concentrating the polynucleic acids present in the sample;
(ii) if need be amplifying the relevant part of the rpoB gene with at least one suitable primer pair as described in claims 22 to 23;
(iii) hybridizing the polynucleic acids of step (i) or (ii) with a set of probes, under appropiate hybridization and wash conditions, with said set comprising at least one of the following rpoB wild-type or S-probes as specified in Table 2:
20 S1 (SEQ ID NO 1), S11 (SEQ ID NO 2), S2 (SEQ ID NO 3), S3 (SEQ ID NO 4), S33 (SEQ ID NO 5), S4 (SEQ ID NO 6), S44 (SEQ ID NO 7), S444 (SEQ ID NO 43), S4444 (SEQ ID NO 8), S5 (SEQ ID NO 9), S55 (SEQ ID NO 10), S555 (SEQ ID NO 39), S5555 (SEQ ID NO 40), S55C (SEQ ID NO 44), 555M (SEQ ID NO 45), S6 (SEQ ID NO 11), and/or. S66 (SEQ ID NO 12),
(iv) detecting the hybrids formed in step (iii);
(v) inferring the rifampicin susceptibility (sensitivity versus resistance) of M. tuberculosis present in the sample from the differential hybridization signal(s) obtained in step (iv).

3 more elements

Set of primers allowing amplification of the rpoB gene fragment of M. tuberculosis, selected from the following sets of primers as specified in Table 2:

31 P1 and P5 (SEQ ID NO 30 and 33), P3 and P4 (SEQ ID NO 31 and 32), P7 and P8 (SEQ ID NO 41 and 42), or. P2 and P6, in combination with (P1 and P5) or (P3 and P4) or (P7 and P8).

Set of primers allowing the amplification of the rpoB gene fragment in mycobacteria other than M. tuberculosis, composed of a 5′ primer selected from

32 MGRPO-1 (SEQ ID NO 64), or, MGRPO-2 (SEQ ID NO 65), and a 3′-primer, selected from:
MGRPO-3 (SEQ ID NO 66), or, MGRPO-4 (SEQ ID NO 67).

Dependencies are inferred from the claim text.

Method for the detection of the antibiotic resistance spectrum of Mycobacterium species

Life of this patent

Method for the detection of the antibiotic resistance spectrum of Mycobacterium species

Life of this patent

Abstract

Claims27