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Rapid diagnostic multiplex PCR (RD-PCR) to discriminate Schistosoma haematobium and S. bovis

Published online by Cambridge University Press:  03 August 2009

B.L. Webster ,
D. Rollinson ,
J.R. Stothard  and
T. Huyse
B.L. Webster*
Affiliation:
Wolfson Wellcome Biomedical Laboratories, Department of Zoology, The Natural History Museum, Cromwell Road, LondonSW7 5BD, UK
D. Rollinson
Affiliation:
Wolfson Wellcome Biomedical Laboratories, Department of Zoology, The Natural History Museum, Cromwell Road, LondonSW7 5BD, UK
J.R. Stothard
Affiliation:
Wolfson Wellcome Biomedical Laboratories, Department of Zoology, The Natural History Museum, Cromwell Road, LondonSW7 5BD, UK
T. Huyse
Affiliation:
Department of Parasitology, Institute of Tropical Medicine, Nationalestraat 155, B-2000Antwerpen, Belgium Laboratory of Animal Diversity and Systematics, Catholic University of Leuven, Ch. de Beriotstraat 32, B-3000Leuven, Belgium
*
*E-mail: [email protected]
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Abstract

Schistosoma haematobium and S. bovis are widespread schistosome species causing human and cattle schistosomiasis, respectively, in Africa. The sympatric occurrence of these two species and their ability to infect the same Bulinus intermediate snail hosts necessitates precise methods of identification of the larval stages. A rapid diagnostic ‘mulitplex’ one-step polymerase chain reaction protocol (RD-PCR) was developed using cytochrome oxidase subunit 1 (COX1) mitochondrial DNA (mtDNA) to discriminate between S. haematobium and S. bovis. A single forward primer and two species-specific reverse primers were used to produce a polymerase chain reaction (PCR) fragment of 306 bp and 543 bp for S. bovis and S. haematobium, respectively. Serial dilutions were carried out on various lifecycle stages and species combinations to test the sensitivity and specificity of the primers. This RD-PCR proved highly sensitive, detecting a single larval stage and as little as 0.78 ng of genomic DNA (gDNA) from an adult schistosome, providing a cost-effective, rapid and robust molecular tool for high-throughput screening of S. haematobium and S. bovis populations. In areas where human and cattle schistosomiasis overlap and are transmitted in close proximity, this mitochondrial assay will be a valuable identification tool for epidemiological studies, especially when used in conjunction with other nuclear diagnostic markers.

Type
Research Papers
Information
Journal of Helminthology , Volume 84 , Issue 1 , March 2010 , pp. 107 - 114
Copyright
Copyright © Cambridge University Press 2009

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