US 12,391,997 B2
Method for determining the presence or absence of M. tuberculosis, M. bovis and M. bovis BCG in a sample
Kwang-Il Lee, Seoul (KR); and Mi Seon Lee, Gyeonggi-do (KR)
Assigned to SEEGENE, INC., Seoul (KR)
Appl. No. 16/959,453
Filed by SEEGENE, INC., Seoul (KR)
PCT Filed Dec. 28, 2018, PCT No. PCT/KR2018/016910
§ 371(c)(1), (2) Date Feb. 25, 2021,
PCT Pub. No. WO2019/135567, PCT Pub. Date Jul. 11, 2019.
Claims priority of application No. 10-2018-0001969 (KR), filed on Jan. 5, 2018.
Prior Publication US 2022/0090170 A1, Mar. 24, 2022
Int. Cl. C12Q 1/689 (2018.01)
CPC C12Q 1/689 (2013.01) [C12Q 2600/16 (2013.01)] 10 Claims
 
1. A method for determining the presence or absence of Mycobacterium tuberculosis, Mycobacterium bovis, and Mycobacterium bovis BCG in a sample comprising nucleic acid molecules, the method comprising:
(a) mixing a sample comprising nucleic acid molecules with a nucleic acid amplification composition comprising (i) a pair of primers for detecting an mpb64 gene; (ii) a pair of primers for detecting an RD9 gene; (iii) a pair of primers for detecting an RD1 gene deletion; and (iv) a pair of primers for detecting an RD4 gene deletion, wherein the pair of primers for detecting the mpb64 gene or the RD9 gene produce an amplicon when the mpb64 gene or the RD9 gene is present, and the pair of primers for detecting the RD1 gene deletion or the RD4 gene deletion produce an amplicon when the RD1 gene deletion or the RD4 gene deletion is present;
(b) performing a nucleic acid amplification reaction;
(c) determining the presence or absence of the mpb64 gene, the presence or absence of the RD9 gene, the presence or absence of the RD1 gene deletion, and the presence or absence of the RD4 gene deletion, from results of the nucleic acid amplification reaction; and
(d) determining the presence or absence of M. tuberculosis, M. bovis, and M. bovis BCG, respectively by using results obtained in step (c).