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Comparative Sensitivity of PCR Primer Sets for Detection of Cryptosporidium parvum

Korean Journal of Parasitology 2009년 47권 3호 p.293 ~ 297
유재란 ( Yu Jae-Ran ) - 건국대학교 의과대학 Department of Environmental and Tropical Medicine

이수웅 ( Lee Soo-Ung ) - 건국대학교 의과대학 Department of Environmental and Tropical Medicine
박우윤 ( Park Woo-Yoon ) - 충북대학교 의과대학 방사선종양학교실

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 103 to 104 oocysts, and the nested PCR method was able to detect 100 to 102 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.

키워드

Cryptosporidium parvum;nested PCR;COWP gene;SSU rRNA
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