Shimada Masamitsu, Hino Fumitsugu, Yamamoto Junko, Mukai Hiroyuki, Hosobe Takahide, Onodera Shoichi, Hoshina Sadayori, Machida Katsuhiko
Takara BIO INC., Otsu 520-2193.
Rinsho Byori. 2003 Nov;51(11):1061-7.
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 simultaneous detection system for Chlamydia trachomatis/Neisseria gonorrhoeae DNA, combined with luminescence detection by a probe hybridization. In the performance tests, this system was able to detect 10 to 100 copies of C. trachomatis/N. gonorrhoeae DNA for only 3.5 hours, and was highly specific to C. trachomatis/N. gonorrhoeae without any cross-reaction to C. pneumoniae, N. lactamica, N. sicca or N. meningitidis. When we tested 60 clinical samples of urine and cervical swabs, the interpretive results were completely consistent with those obtained by Roche PCR system. Of 13 positive samples by the ICAN and PCR systems for C. trachomatis, four were negative by EIA method(IDEIA Chlamydia). These results indicate that the ICAN system is an efficient and sensitive system to simultaneously detect C. trachomatis/N. gonorrhoeae DNA.
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、核糖核酸酶H和DNA-RNA嵌合引物组成。我们开发了沙眼衣原体/淋病奈瑟菌DNA同时检测系统,并结合探针杂交进行发光检测。在性能测试中,该系统仅需3.5小时就能检测到10至100拷贝的沙眼衣原体/淋病奈瑟菌DNA,对沙眼衣原体/淋病奈瑟菌具有高度特异性,对肺炎衣原体、乳糖奈瑟菌、微黄奈瑟菌或脑膜炎奈瑟菌无任何交叉反应。当我们检测60份尿液和宫颈拭子临床样本时,解释结果与罗氏PCR系统获得的结果完全一致。在ICAN和PCR系统检测为沙眼衣原体阳性的13份样本中,有4份经酶免疫分析法(IDEIA衣原体)检测为阴性。这些结果表明,ICAN系统是一种高效、灵敏的同时检测沙眼衣原体/淋病奈瑟菌DNA的系统。