Detection of mycobacteria by multiplex nucleic acid amplification

US5811269A · kind A · utility

Assignee

• Becton, Dickinson and Company · US

Inventors

• James G. Nadeau · Chapel Hill, US
• Cheryl Dean · Raleigh, US
• James L. Schram · Knightdale, US
• Deborah R. Howard · Durham, US
• Margaret S. Dey · Apex, US
• David J. Wright · Chapel Hill, US

Key dates

• Filing date: Apr 30, 1996
• Grant date: Sep 22, 1998
• Expiry date: Apr 30, 2016

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12Q2600/16
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

Primers and methods for adapter-mediated multiplex amplification of the IS6110 insertion element of the Mycobacterium tuberculosis (M.tb) complex and a 16S rDNA target common to essentially all mycobacteria are described. In certain embodiments, the primers are optimized for efficient multiplex amplification in thermophilic SDA. The multiplex Strand Displacement Amplification methods of the invention are capable, in a single amplification reaction, of simultaneously identifying M. tuberculosis and providing a screen for substantially all of the clinically relevant species of mycobacteria. Also disclosed are internal control sequences designed for coamplification with the two targets, allowing assessment of amplification efficiency and/or quantitation of the targets.