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

Li, Juan, Zhao, Guang-Hui, Lin, RuiQing, Blair, David, Sugiyama, Hiromu, and Zhu, Xing-Quan (2015) Rapid detection and identification of four major Schistosoma species by high-resolution melt (HRM) analysis. Parasitology Research, 114 (11). pp. 4225-4232.

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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 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.

Item ID: 41824
Item Type: Article (Research - C1)
ISSN: 1432-1955
Keywords: schistosoma, schistosomiasis, high-resolution meltcurve (HRM), rapididentification, differentiation, 18S rDNA
Funders: Special Fund for Agro-Scientific Research in the Public Interest (SFASR), Science Fund for Creative Research Groups of Gansu Province (SFCRGG), Pearl River Science and Technology (PRST), Guangdong Academy of Agricultural Sciences (GAAS)
Projects and Grants: SFASR Grant No. 201303037, SFCRGG Grant No. 120RJIA006, PRST New Star Project Grant No. 2014 J2200096, GAAS Grant No. 201413
Date Deposited: 08 Dec 2015 18:41
FoR Codes: 11 MEDICAL AND HEALTH SCIENCES > 1108 Medical Microbiology > 110803 Medical Parasitology @ 100%
SEO Codes: 92 HEALTH > 9204 Public Health (excl. Specific Population Health) > 920499 Public Health (excl. Specific Population Health) not elsewhere classified @ 100%
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