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[基于等温DNA扩增法ICAN的结核分枝杆菌DNA检测系统的研制]

[Development of the detection system for Mycobacterium tuberculosis DNA by using the isothermal DNA amplification method ICAN].

作者信息

Shimada Masamitsu, Hino Fumitsugu, Sagawa Hiroaki, Mukai Hiroyuki, Asada Kiyozo, Kato Ikunoshin

机构信息

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

出版信息

Rinsho Byori. 2002 May;50(5):528-32.

Abstract

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.

摘要

核酸等温嵌合引物引发扩增(ICAN)是一种新的等温DNA扩增方法,由外切Bca DNA聚合酶、RNaseH和DNA-RNA嵌合引物组成。我们开发了一种检测系统,将ICAN与通过探针杂交的发光检测相结合,用于靶向IS6110插入元件的结核分枝杆菌DNA检测。我们对该系统进行了性能测试。该系统仅需3.5小时就能检测到一份结核分枝杆菌DNA,其性能与罗氏PCR系统相当或更优。我们还研究了一种使用磁珠的检测系统,该系统可将检测时间缩短至2.5小时。结果表明,ICAN系统是一种用于从大量样本中检测结核分枝杆菌DNA的高效且灵敏的检测系统。

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