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Korean J Parasitol. 2009 Sep;47(3):293-297.
Published online 2009 August 28.  doi: 10.3347/kjp.2009.47.3.293.
Copyright © 2009 by The Korean Society for Parasitology
Comparative Sensitivity of PCR Primer Sets for Detection of Cryptosporidium parvum
Jae-Ran Yu,1 Soo-Ung Lee,1 and Woo-Yoon Park2
1Department of Environmental and Tropical Medicine, Konkuk University, School of Medicine, Seoul 143-701, Korea.
2Department of Radiation Oncology, College of Medicine, Chungbuk National University, Cheongju 361-763, Korea.

Corresponding author (Email: maria205@kku.ac.kr )
Received February 24, 2009; Revised April 22, 2009; Accepted May 16, 2009.

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.

Keywords: Cryptosporidium parvum, nested PCR, COWP gene, SSU rRNA.
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