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

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

The Korean Journal of Parasitology 2013;51(6):651-656.
Published online: December 31, 2013

1Department of Parasitology, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand.

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6Applied Malacology Center, Department of Social and Environmental Medicine, Faculty of Tropical Medicine, Mahidol University, Bangkok 10400, Thailand.

Corresponding author (wanch_ma@kku.ac.th)
• Received: May 30, 2013   • Revised: August 7, 2013   • Accepted: October 11, 2013

© 2013, Korean Society for Parasitology and Tropical Medicine

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Molecular Differentiation of Schistosoma japonicum and Schistosoma mekongi by Real-Time PCR with High Resolution Melting Analysis
Korean J Parasitol. 2013;51(6):651-656.   Published online December 31, 2013
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Molecular Differentiation of Schistosoma japonicum and Schistosoma mekongi by Real-Time PCR with High Resolution Melting Analysis
Korean J Parasitol. 2013;51(6):651-656.   Published online December 31, 2013
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Molecular Differentiation of Schistosoma japonicum and Schistosoma mekongi by Real-Time PCR with High Resolution Melting Analysis
Image Image Image
Fig. 1 (A) Representative melting peaks (℃) for Schistosoma japonicum (a), Schistosoma mekongi (b), mixed-plasmids (c), and distilled water (d). Amplification plot of fluorescence vs cycle number showing analytical sensitivity of HRM real-time PCR for detection of S. japonicum (B) and S. mekongi (C) plasmids: e-j; 10-fold serial dilutions of S. japonicum or S. mekongi plasmids, from 4.3×107 to 4.3 ×102 copies per reaction. k; distilled water (negative control).
Fig. 2 Analytical sensitivity for detection of cercariae (A, B) and eggs (C, D) of S. japonicum (A, C) and S. mekongi (B, D). Cycle numbers for detection of 4 cercariae (l), 2 cercariae (m), 1 cercaria (n), and for detection of 8 eggs (p), 4 eggs (q), 2 eggs (r), and 1 egg (s). o and t; distilled water (negative control).
Fig. 3 Ethidium bromide staining patterns of the PCR products on a 1.5% agarose gel. The arrows indicate the 156 bp of S. mekongi and S. japonicum specific bands. Lane M: DNA size markers (1 kb plus DNA ladder from Invitrogen, Carlsbad, California, USA). Negative control containing no DNA (Lane 1); S. mekongi positive control plasmid (Lane 2); S. japonicum positive control plasmid (Lane 3); S. mekongi-infected Neotricula aperta snails (Lane 4); non-infected N. aperta snails (Lane 5); S. japonicum-infected Oncomelania nosophora snails (Lane 6); non-infected O. nosophora snails (Lane 7); S. mekongi-infected rat feces (Lane 8); negative healthy human feces (Lane 9); and S. japonicum-infected mice feces (Lane 10).
Molecular Differentiation of Schistosoma japonicum and Schistosoma mekongi by Real-Time PCR with High Resolution Melting Analysis
Cycle numbers
Melting temperatures
Range Mean ± SD Median Range Mean ± SD Median S. japonicum-infected mice (n = 9) 15.8-29.7 22.0 ± 4.2 21.9 84.4-84.6 84.5 ± 0.07 84.5 S. mekongi-infected rats (n = 12) 19.5-29.2 23.5 ± 3.4 22.6 85.6-85.7 85.7 ± 0.04 85.7
Table 1. The cycle number and melting temperature values of HRM real-time PCR