用于检测微小隐孢子虫的聚合酶链反应引物组的比较敏感性
Comparative sensitivity of PCR primer sets for detection of Cryptosporidium parvum.
作者信息
Yu Jae-Ran, Lee Soo-Ung, Park Woo-Yoon
机构信息
Department of Environmental and Tropical Medicine, Konkuk University, School of Medicine, Seoul, Korea.
出版信息
Korean J Parasitol. 2009 Sep;47(3):293-7. doi: 10.3347/kjp.2009.47.3.293. Epub 2009 Aug 28.
Abstract
Improved methods for detection of Cryptosporidium oocysts in environmental and clinical samples are urgently needed to improve detection of cryptosporidiosis. We compared the sensitivity of 7 PCR primer sets for detection of Cryptosporidium parvum. Each target gene was amplified by PCR or nested PCR with serially diluted DNA extracted from purified C. parvum oocysts. The target genes included Cryptosporidium oocyst wall protein (COWP), small subunit ribosomal RNA (SSU rRNA), and random amplified polymorphic DNA. The detection limit of the PCR method ranged from 10(3) to 10(4) oocysts, and the nested PCR method was able to detect 10(0) to 10(2) oocysts. A second-round amplification of target genes showed that the nested primer set specific for the COWP gene proved to be the most sensitive one compared to the other primer sets tested in this study and would therefore be useful for the detection of C. parvum.
摘要
迫切需要改进环境和临床样本中隐孢子虫卵囊的检测方法，以提高隐孢子虫病的检测水平。我们比较了7种用于检测微小隐孢子虫的PCR引物组的灵敏度。通过PCR或巢式PCR对从纯化的微小隐孢子虫卵囊中提取的系列稀释DNA进行各靶基因的扩增。靶基因包括隐孢子虫卵囊壁蛋白（COWP）、小亚基核糖体RNA（SSU rRNA）和随机扩增多态性DNA。PCR方法的检测限为10³至10⁴个卵囊，巢式PCR方法能够检测10⁰至10²个卵囊。靶基因的第二轮扩增表明，与本研究中测试的其他引物组相比，针对COWP基因的巢式引物组被证明是最灵敏的，因此可用于微小隐孢子虫的检测。
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