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Rapid detection and identification of four major Schistosoma species by high-resolution melt (HRM) analysis

文献类型： 外文期刊

作者： Li, Juan 1 ; Zhao, Guang-Hui 3 ; Lin, RuiQing 4 ; Blair, David 5 ; Sugiyama, Hiromu 6 ; Zhu, Xing-Quan 1 ;

作者机构： 1.Chinese Acad Agr Sci, Lanzhou Vet Res Inst, Key Lab Vet Parasitol Gansu Prov, State Key Lab Vet Etiol Biol, Lanzhou 730046, Gansu, Peoples R China

2.Guangdong Acad Agr Sci, Inst Anim Hlth, Guangzhou 510640, Guangdong, Peoples R China

3.Northwest A&F Univ, Coll Vet Med, Yangling 712100, Shaanxi Provinc, Peoples R China

4.South China Agr Univ, Coll Vet Med, Guangzhou 510642, Guangdong, Peoples R China

5.James Cook Univ, Sch Marine & Trop Biol, Townsville, Qld 4811, Australia

6.Natl Inst Infect Dis, Dept Parasitol, Tokyo 1138421, Japan

关键词： Schistosoma;Schistosomiasis;High-resolution meltcurve( HRM);Rapididentification;Differentiation;18S rDNA

期刊名称：PARASITOLOGY RESEARCH （ 影响因子：2.289； 五年影响因子：2.403 ）

ISSN：

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收录情况： SCI

摘要： Schistosomiasis, caused by blood flukes belonging to several species of the genus Schistosoma, is a serious and widespread parasitic disease. Accurate and rapid differentiation of these etiological agents of animal and human schistosomiasis to species level can be difficult. We report a real-time PCR assay coupled with a high-resolution melt (HRM) assay targeting a portion of the nuclear 18S rDNA to detect, identify, and distinguish between four major blood fluke species (Schistosoma japonicum, Schistosoma mansoni, Schistosoma haematobium, and Schistosoma mekongi). Using this system, the Schistosoma spp. was accurately identified and could also be distinguished from all other trematode species with which they were compared. As little as 10(-5) ng genomic DNA from a Schistosoma sp. could be detected. This process is inexpensive, easy, and can be completed within 3 h. Examination of 21 representative Schistosoma samples from 15 geographical localities in seven endemic countries validated the value of the HRM detection assay and proved its reliability. The melting curves were characterized by peaks of 83.65 A degrees C for S. japonicum and S. mekongi, 85.65 A degrees C for S. mansoni, and 85.85 A degrees C for S. haematobium. The present study developed a real-time PCR coupled with HRM analysis assay for detection and differential identification of S. mansoni, S. haematobium, S. japonicum, and S. mekongi. This method is rapid, sensitive, and inexpensive. It has important implications for epidemiological studies of Schistosoma.

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