Korean J Parasitol.  2009 Sep;47(3):293-297. 10.3347/kjp.2009.47.3.293.

Comparative Sensitivity of PCR Primer Sets for Detection of Cryptosporidium parvum

• Yu JR
• Lee SU
• Park WY

• KMID: 1729463
• DOI: http://doi.org/10.3347/kjp.2009.47.3.293

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.

Keyword

Cryptosporidium parvum; nested PCR; COWP gene; SSU rRNA

MeSH Terms

Animals
Cryptosporidiosis/diagnosis/*parasitology
Cryptosporidium parvum/genetics/*isolation & purification
DNA Primers/*genetics
DNA, Protozoan/diagnostic use/genetics
Humans
Polymerase Chain Reaction/*methods
Sensitivity and Specificity