[Rinsho Byori 50 : 528〜532, 2002]

Development of the Detection System for Mycobacterium tuberculosis DNA
by Using the Isothermal DNA Amplification Method ICAN

Masamitsu SHIMADA, PhD*1, Fumitsugu HINO, PhD*2, Hiroaki SAGAWA, PhD*3,
Hiroyuki MUKAI*4, Kiyozo ASADA, PhD*5 and Ikunoshin KATO, PhD*6

The isothermal and chimeric primer-initiated amplification of nucleic acids(ICAN) is a new isothermal DNA amplification method composed of exo- Bca DNA polymerase, RNaseH and DNA-RNA chimeric primers. We developed the detection system, combined the ICAN with luminescence detection by a probe hybridization, for Mycobacterium tuberculosis DNA targeting the IS6110 insertion element. We examined performance tests of the system. This system was able to detect one copy of Mycobacterium tuberculosis DNA for only 3.5 hours, and performance of the system was equivalent or better to the Roche PCR system. We also examined a detection system by using magnetic beads, which system could shorten detection time for 2.5 hours. It was shown that the ICAN system was an efficient and sensitive detection system for Mycobacterium tuberculosis DNA from mass samples.


*1Biomedical Group, Takara Shuzo Co. Ltd., Otsu 520-2193

【Key Words】Mycobacterium tuberculosis, isothermal and chimeric primer-initiated amplification of nucleic acid : ICAN, gene amplification, IS6110