Mayo Foundation for Medical Education and Research

Roche Molecular Systems, Inc.

Chiron Diagnostic Corporation

Glenn D Roberts

Gurpreet S Sandhu

Karen K Young

Bruce C Kline

Marcia E Lewis

David H Persing

John J Hunt

Leslie Stockman

A Christian Whelan

Teresa A Felmlee

The detection of the IS6110 insertion element in a clinical specimen is a rapid way of diagnosing infection by Mycobacterium tuberculosis. A reliable diagnostic test for tuberculosis based on the IS6110 DNA is described in this disclosure. A 'Universal' specimen preparation protocol that eliminates live organisms and purifies nucleic acids from all types of clinical specimens is described. Two nucleic acid primers designed to amplify IS6110 DNA with high specificity in a polymerase chain reaction are also described. The amplified IS6110 DNA is identified by a restriction endonuclease and electrophoresis based assay. The identification process also renders the DNA unamplifiable in a subsequent PCR, thereby reducing the possibility of contaminating other specimens. Time, labor and cost is minimized, while user safety and test reliability are maximized. The complete DNA extraction, amplification and analysis is accomplished with ease within an 8 hour period, with a sensitivity of 92% and a specificity approaching 100%. Testing of serially obtained samples from the same patient increases the overall rate of detection to 100%.

IS6110 based molecular detection of mycobacterium tuberculosis

A method is provided for detecting M. tuberculosis or mutants thereof, particularly rifampin-resistant MTB, in a biological sample comprising: isolating DNA from the biological sample; amplifying the isolated DNA under hybridizing conditions with a primer set that targets portions of the gene encoding rpoB; wherein the pimer set comprises at least one primer that hybridizes under hybridizing conditions to at least one signature nucleotide for M. tuberculosis; and isolating and sequencing the amplified DNA to determine the presence or absence of M. tuberculosis or mutants thereof.

Detection of a genetic locus encoding resistance to rifampin in

Growing community of inventors

Rochester, MN, United States of America

Glenn D Roberts