Specific PCR-based assays for the identification of Fasciola species: Their development, evaluation and potential usefulness in prevalence surveys

L. Ai, S. J. Dong, W. Y. Zhang, H. M. Elsheikha, Y. S. Mahmmod, R. Q. Lin, Z. G. Yuan, Y. L. Shi, W. Y. Huang, X. Q. Zhu

Research output: Contribution to journalArticlepeer-review

44 Scopus citations

Abstract

Among the helminths infecting ruminants in China are three taxa belonging to the genus Fasciola: F. hepatica, F. gigantica and the so-called 'intermediate form' that appears to lie between these two species. Based on the sequences of the second internal-transcribed spacers (ITS-2) within the parasites' nuclear ribosomal DNA (rDNA), a pair of primers (DSJf/DSJ3) specific for F. hepatica and a pair (DSJf/DSJ4) specific for F. gigantica were designed and used to develop PCR-based assays. These assays allowed the identification and differentiation of F. hepatica, F. gigantica and the 'intermediate' Fasciola, with no amplicons produced from heterologous DNA samples. The results of sequencing confirmed the species-specific identity of the amplified products. The assays showed good sensitivity, giving positive results with as little as 0.11 ng of F. hepatica DNA and 0.35 ng of F. gigantica DNA. This meant that the DNA from a single Fasciola egg or a single infected snail was sufficient for identification of the Fasciola taxon. The developed PCR assays could provide useful tools for the detection, identification and epidemiological investigation of Fasciola infection in humans, other mammals and snails.

Original languageEnglish (US)
Pages (from-to)65-72
Number of pages8
JournalAnnals of Tropical Medicine and Parasitology
Volume104
Issue number1
DOIs
StatePublished - Jan 2010

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