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

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Abstract

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 °C for S. japonicum and S. mekongi, 85.65 °C for S. mansoni, and 85.85 °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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Acknowledgments

Project support was provided by the “Special Fund for Agro-scientific Research in the Public Interest” (Grant No. 201303037) and the Science Fund for Creative Research Groups of Gansu Province (Grant No. 1210RJIA006) to XQZ, and the Pearl River Science and Technology New Star Project (Grant No. 2014 J2200096) and the President’s Foundation of Guangdong Academy of Agricultural Sciences (Grant No. 201413) to JL.

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Authors and Affiliations

  1. State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu Province, 730046, People’s Republic of China

    Juan Li & Xing-Quan Zhu

  2. Institute of Animal Health, Guangdong Academy Agricultural Sciences, Guangzhou, Guangdong Province, 510640, People’s Republic of China

    Juan Li

  3. College of Veterinary Medicine, Northwest A & F University, Yangling, Shaanxi Province, 712100, People’s Republic of China

    Guang-Hui Zhao

  4. College of Veterinary Medicine, South China Agricultural University, Guangzhou, Guangdong Province, 510642, People’s Republic of China

    RuiQing Lin

  5. School of Marine and Tropical Biology, James Cook University, Townsville, Queensland, 4811, Australia

    David Blair

  6. Department of Parasitology, National Institute of Infectious Diseases, 113-8421, Tokyo, Japan

    Hiromu Sugiyama

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  1. Juan Li
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  2. Guang-Hui Zhao
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  4. David Blair
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  6. Xing-Quan Zhu
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Correspondence to Xing-Quan Zhu.

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Li, J., Zhao, GH., Lin, R. et al. Rapid detection and identification of four major Schistosoma species by high-resolution melt (HRM) analysis. Parasitol Res 114, 4225–4232 (2015). https://doi.org/10.1007/s00436-015-4660-3

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  • DOI: https://doi.org/10.1007/s00436-015-4660-3

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