Mutation detection on RNA polmerase beta subunit gene having rifampin resistance

US6902894B2 · kind B2 · utility

Assignee

• GENEREX PHARMACEUTICALS, INC. · CA

Inventors

• Mengsu YANG · Hong Kong, CN
• Hok Sin Woo · Hong Kong, CN

Key dates

• Filing date: Sep 7, 2001
• Grant date: Jun 7, 2005
• Expiry date: Apr 3, 2022

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12Q2600/156
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

The present invention provides a diagnostic test method for detecting a tendency to rifampin resistance caused by mutations in a rpoB gene of M. tuberculosis, comprising the steps of (i) extracting genomic DNA from a biological sample containing M. tuberculosis cells; (ii) amplifying from the extracted genomic DNA the rpoB gene coding sequence or at least one distinct fragment thereof containing nucleotides encoding at least one test amino acid of the group consisting of amino acid numbers 511, 512, 513, 514, 515, 516, 517, 518, 522, 526, 529, 531, 533 to produce fluorescently labeled amplification product; (iii) contacting said fluorescently labeled amplification product with a first control array of oligonucleotide probes having DNA sequences specific to the wildtype M. tuberculosis rpoB gene coding sequence, including the nucleotides encoding the at least one test amino acid, and with a second test array of oligonucleotide probes having DNA sequences specific to the M. tuberculosis rpoB gene coding sequence, including nucleotides encoding mutations in the at least one test amino acid, wherein at least 3 mutations of the rpoB gene are probed for by the second test array of oligon…