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Korean J Parasitol. 2013 Dec;51(6):651-6. doi: 10.3347/kjp.2013.51.6.651. Epub 2013 Dec 31.

Molecular differentiation of Schistosoma japonicum and Schistosoma mekongi by real-time PCR with high resolution melting analysis.

Author information

1
Department of Parasitology, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand. ; Research and Diagnostic Center for Emerging Infectious Diseases, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand.
2
Research and Diagnostic Center for Emerging Infectious Diseases, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand. ; Department of Biochemistry, Faculty of Medical Science, Naresuan University, Phitsanulok, 65000 Thailand.
3
Research and Diagnostic Center for Emerging Infectious Diseases, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand. ; Faculty of Medicine, Mahasarakham University, Mahasarakham 44000, Thailand.
4
Research and Diagnostic Center for Emerging Infectious Diseases, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand. ; Department of Microbiology, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand.
5
Applied Malacology Center, Department of Social and Environmental Medicine, Faculty of Tropical Medicine, Mahidol University, Bangkok 10400, Thailand.

Abstract

Human schistosomiasis caused by Schistosoma japonicum and Schistosoma mekongi is a chronic and debilitating helminthic disease still prevalent in several countries of Asia. Due to morphological similarities of cercariae and eggs of these 2 species, microscopic differentiation is difficult. High resolution melting (HRM) real-time PCR is developed as an alternative tool for the detection and differentiation of these 2 species. A primer pair was designed for targeting the 18S ribosomal RNA gene to generate PCR products of 156 base pairs for both species. The melting points of S. japonicum and S. mekongi PCR products were 84.5±0.07℃ and 85.7±0.07℃, respectively. The method permits amplification from a single cercaria or an egg. The HRM real-time PCR is a rapid and simple tool for differentiation of S. japonicum and S. mekongi in the intermediate and final hosts.

KEYWORDS:

Schistosoma japonicum; Schistosoma mekongi; differentiation; high resolution melting analysis; real-time PCR

PMID:
24516269
PMCID:
PMC3916453
DOI:
10.3347/kjp.2013.51.6.651
[Indexed for MEDLINE]
Free PMC Article

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