Korean J Parasitol.  2013 Dec;51(6):651-656. 10.3347/kjp.2013.51.6.651.

Molecular Differentiation of Schistosoma japonicum and Schistosoma mekongi by Real-Time PCR with High Resolution Melting Analysis

Affiliations
  • 1Department of Parasitology, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand. wanch_ma@kku.ac.th
  • 2Research and Diagnostic Center for Emerging Infectious Diseases, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand.
  • 3Department of Biochemistry, Faculty of Medical Science, Naresuan University, Phitsanulok, 65000 Thailand.
  • 4Faculty of Medicine, Mahasarakham University, Mahasarakham 44000, Thailand.
  • 5Department of Microbiology, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand.
  • 6Applied 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+/-0.07degrees C and 85.7+/-0.07degrees C, 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.

Keyword

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

MeSH Terms

Animals
DNA Primers/genetics
Mice
Parasitology/*methods
RNA, Ribosomal, 18S/genetics
Real-Time Polymerase Chain Reaction/*methods
Schistosoma/*classification/*genetics
Snails
Time Factors
Transition Temperature
DNA Primers
RNA, Ribosomal, 18S
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