US 6815165 B2 - Method For Identifying Mycobacterium Tuberculosis And Mycobacteria Other Than Tuberculosis, Together With Detecting Resistance To An Antituberculosis Drug Of Mycobacteria Obtained By Mutation Of Rpob Gene - The Lens - Free & Open Patent and Scholarly Search

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US 6815165 B2

Abstract

The present invention provides a method for identifying Mycobacterium tuberculosis and non-tuberculosis Mycobacterium (MOTT), and for the determination of drug susceptibility of M. tuberculosis based on detection of mutations in the rpoB gene.

Claims

A method for identifying Mycobacterium tuberculosis and non-tuberculosis Mycobacterium (MOTT) which comprises the following steps:
(1) isolating DNA from a sample;
(2) amplifying a 531 bp fragment of the rpoB gene by PCR using said DNA isolated in the above step (1) and the primers of SEQ. ID. NOs. 1 and 2;
(3) performing PCR-reverse blot hybridization by hybridizing the PCR product obtained in the above step (2) with one or more oligomer probes, said oligomer probes selected from the group of oligomer probes consisting of SEQ. ID. NOs. 5, 6, 9, 10, 13 and 14, adhered to a suitable membrane.
The method of claim 1 , wherein said suitable membrane is a negatively-charged nylon membrane.
The method of claim 2 , wherein said negatively-charged nylon membrane has surface carboxyl groups.
A pair of oligonucleotide primers consisting of the DNA sequences described by SEQ. ID. NOs. 1 and 2, respectively.
An oligomer probe selected from the group consisting of SEQ. ID. NOs. 5, 6, 9, 10, 13 and 14, wherein said probe can be used to hybridize to a PCR-amplified fragment of the mycobacterial rpoB gene in order to separately identify Mycobacterium tuberculosis and MOTTs.
A kit for separately identifying Mycobacterium tuberculosis and MOTTs, which comprises (1) a pair of oligonucleotide primers described by SEQ. ID. NOs. 1 and 2 and (2) one or more oligomer probes selected from the group consisting of SEQ. ID. NOs. 5, 6, 9, 10, 13 and 14.

Owners (US)

Cho Sang-nae (Jul 10 2003)
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Lee Hyeyoung (Jul 10 2003)
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Xeniss Life Science Co. Ltd (Dec 18 2001)
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Applicants

Cho Sang-nae
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Lee Hyeyoung
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Inventors

Lee Hyeyoung
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Bang Hye Eun
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Cho Sang-nae
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Bai Gill-han
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Kim Sang-jae
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CPC Classifications

C12Q1/689
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C12Q2600/156
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Y10S435/81
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IPC Classifications

C12Q1/68
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C12Q1/689
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US Classifications

435/6
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435/91.2
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435/810
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536/22.1
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536/24.33
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Document History

Publication: Nov 9, 2004
US 6815165 B2
Application: Jan 30, 2002
US 5842202 A
Priority: Jul 19, 2001
KR 20010043450 A

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