Back to results
US 7,329,493 B2Granted
One-tube nested PCR for detecting Mycobacterium tuberculosis
AssigneeAsiaGEN Corporation
InventorsGeorge Chin-Sheng Chou, Kai-Pin Huang
Life of this patent
Estimating the term…
Abstract
The present invention relates to a kit and method for detecting M. tuberculosis of suspected patient. The present invention also relates to primers and probe used to detect M. tuberculosis by performing one tube nested PCR.
Claims17
A kit for detecting the presence or absence of Mycobacterium tuberculosis in a sample using a nested polymerase chain reaction, comprising
- (I) an outer pair of oligonucleotide primers selected from the group consisting of (a) SEQ ID NOS:
- 2 and 4, and (b) SEQ ID NOS:
- 3 and 5, and
- (ii) an inner pair of oligonucleotide primers selected from the group consisting of (c) SEQ ID NOS:
- 6 and 8, and (d) SEQ ID NOS:
- 7 and 9.
3 more elements
A method for detecting the presence or absence of Mycobacterium tuberculosis in a sample using a nested polymerase chain reaction, comprising
- (I) adding into one tube with DNA from the sample, buffer, dNTP, Taq polymerase, probe having SEQ ID NO:
- 10, an outer pair of oligonucleotide primers selected from the group consisting of (a) SEQ ID NOS:
- 2 and 4, and (b) SEQ ID NOS:
- 3 and 5, and an inner pair of oligonucleotide primers selected from the group consisting of (c) SEQ ID NOS:
- 6 and 8, and (d) SEQ ID NOS:
- 7 and 9 to form a mixture, wherein the concentration of the outer pair of oligonucleotide primers to the concentration of the inner pair of oligonucleotide primers is 1:5 o 1:100;
- (ii) performing first 20-50 cycles of amplification for PCR products from the outer primers in the first-stage polymerase chain reaction;
- (iii) performing following 10-30 cycles of amplification PCR products from for the inner primers in the second-stage polymerase chain reaction;
- and
- (iv) identifying M. tuberculosis by the probe.
7 more elements
Dependencies are inferred from the claim text.