Michelle L Nelson

US Patent:

6379898, Apr 30, 2002

Filed:

Jan 9, 2001

Appl. No.:

09/757132

Inventors:

John W. Shultz - Verona WI 53593
Michelle A. Nelson - Fitchburg WI 53713
Donna M. Leippe - Madison WI 53705
Martin K. Lewis - Madison WI 53716
Lisa S. Nelson - DeForest WI 53532

International Classification:

C12Q 168

US Classification:

435 6, 435 71

Abstract:

This invention discloses methods, compositions and kits for the detection of extremely low levels of nucleic acid, cells and cellular material in biological samples. The nucleic acid detection systems utilize either the pyrophosphorolysis reaction catalyzed by various polymerases or nuclease digestion coupled with pyrophosphorylation catalyzed by phosphoribosylpyrophosphate synthetase to produce either deoxyribonucleoside triphosphates or ribonucleoside triphosphates. dNTPs are transformed to ATP by the action of nucleoside diphosphate kinase. The ATP produced by these reactions may be detected by luciferase or NADH based detection systems. If more sensitive detection is required, schemes for the amplification of NTPs and dNTPS are provided. A detection system for cells or cellular material in a sample is provided wherein AMP and a high energy phosphate donor added to a sample are converted to ATP by the action of endogenous enzymes, followed by detection of the ATP.

US Patent:

6335162, Jan 1, 2002

Filed:

Mar 13, 1998

Appl. No.:

09/042287

Inventors:

John W. Shultz - Verona WI
Michelle A. Nelson - Fitchburg WI
Donna M. Leippe - Madison WI
Martin K. Lewis - Madison WI
Lisa S. Nelson - DeForest WI

Assignee:

Promega Corporation - Madison WI

International Classification:

C12Q 168

US Classification:

435 6, 435 911

Abstract:

This invention discloses methods, compositions and kits for the detection of extremely low levels of nucleic acid, cells and cellular material in biological samples. The nucleic acid detection systems utilize either the pyrophosphorolysis reaction catalyzed by various polymerases or nuclease digestion coupled with pyrophosphorylation catalyzed by phosphoribosylpyrophosphate synthetase to produce either deoxyribonucleoside triphosphates or ribonucleoside triphosphates. dNTPs are transformed to ATP by the action of nucleoside diphosphate kinase. The ATP produced by these reactions may be detected by luciferase or NADH based detection systems. If more sensitive detection is required, schemes for the amplification of NTPs and dNTPS are provided. A detection system for cells or cellular material in a sample is provided wherein AMP and a high energy phosphate donor added to a sample are converted to ATP by the action of endogenous enzymes, followed by detection of the ATP.