Molecular Differentiation of Schistosoma japonicum and Schistosoma mekongi by Real-Time PCR with High Resolution Melting Analysis.
10.3347/kjp.2013.51.6.651
- Author:
Amornmas KONGKLIENG
1
;
Worasak KAEWKONG
;
Pewpan M INTAPAN
;
Oranuch SANPOOL
;
Penchom JANWAN
;
Tongjit THANCHOMNANG
;
Viraphong LULITANOND
;
Pusadee SRI-AROON
;
Yanin LIMPANONT
;
Wanchai MALEEWONG
Author Information
1. Department of Parasitology, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand. wanch_ma@kku.ac.th
- Publication Type:Original Article ; Evaluation Studies ; Research Support, Non-U.S. Gov't
- Keywords:
Schistosoma japonicum;
Schistosoma mekongi;
differentiation;
high resolution melting analysis;
real-time PCR
- MeSH:
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
- From:The Korean Journal of Parasitology
2013;51(6):651-656
- CountryRepublic of Korea
- Language:English
-
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.
