Deborah D Howard

US Patent:

58112693, Sep 22, 1998

Filed:

Apr 30, 1996

Appl. No.:

8/640378

Inventors:

James G. Nadeau - Chapel Hill NC
Cheryl H. Dean - Raleigh NC
James L. Schram - Knightdale NC
Deborah R. Howard - Durham NC
Margaret S. Dey - Durham NC
David J. Wright - Chapel Hill NC

Assignee:

Becton, Dickinson and Company - Franklin Lakes NJ

International Classification:

C12P 1934
C07H 2104

US Classification:

435 911

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.

US Patent:

55545033, Sep 10, 1996

Filed:

Aug 4, 1994

Appl. No.:

8/286100

Inventors:

James A. Down - Cary NC
Adriann H. Walters - Durham NC
Margaret S. Dey - Apex NC
Deborah R. Howard - Durham NC
Michael C. Little - Raleigh NC
William E. Keating - Durham NC

Assignee:

Becton, Dickinson and Company - Franklin Lakes NJ

International Classification:

C12Q 168
C12P 1934

US Classification:

435 6

Abstract:

Methods for processing samples which may contain mycobacteria which are compatible with both conventional culturing techniques and nucleic acid analysis. It has been found that the harsh chemical treatment previously thought necessary to obtain efficient lysis can be eliminated without loss of lysis efficiency, thereby eliminating reagents which may inhibit subsequent nucleic acid-based reactions. Heat alone is sufficient to lyse mycobacteria, even in buffers which do not contain detergents, chelators, enzymes, etc. Residual reagents introduced by conventional sample processing methods for culture can be successfully removed from the processed pellet without significant loss of microorganisms by at least two washings with saline, water, or buffers compatible with nucleic acid analysis and appropriate centrifugation.