Development of novel multiplex microsatellite polymerase chain reactions to enable high-throughput population genetic studies of Schistosoma haematobium.
BACKGROUND
Human urogenital schistosomiasis caused by Schistosoma haematobium is widely distributed across Africa and is increasingly targeted for control and regional elimination. The development of new high-throughput, cost-effective molecular tools and approaches are needed to monitor and evaluate the impact of control programs on the parasite populations. Microsatellite loci are genetic markers that can be used to investigate how parasite populations change over time and in relation to external influences such as control interventions.


RESULTS
Here, 18 existing S. haematobium microsatellite loci were optimised to enable simultaneous amplification across two novel multiplex microsatellite PCR's, each containing nine loci. Methods were developed for the cost effective and rapid processing and microsatellite analysis of S. haematobium larval stages stored on Whatman-FTA cards and proved robust on miracidia and cercariae collected from Zanzibar and Niger.


CONCLUSIONS
The development of these novel and robust multiplex microsatellite assays, in combination with an improved protocol to elute gDNA from Whatman-FTA fixed schistosome larval stages, enables the high-throughput population genetic analysis of S. haematobium. The molecular resources and protocols described here advance the way researchers can perform multi locus-based population genetic analyses of S. haematobium as part of the evaluation and monitoring of schistosomiasis control programmes.
Wolfson Wellcome Biomedical Laboratories, Department of Life Sciences, Natural History Museum, Cromwell Road, London, SW7 5BD, UK. b.webster@nhm.ac.uk.|Department of Infectious Disease Epidemiology, Imperial College Faculty of Medicine (St Mary's Campus), Norfolk Place, London, W2 1PG, UK. b.webster@nhm.ac.uk.
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Webster
Wolfson Wellcome Biomedical Laboratories, Department of Life Sciences, Natural History Museum, Cromwell Road, London, SW7 5BD, UK. m.rabone@nhm.ac.uk.
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Rabone
Wolfson Wellcome Biomedical Laboratories, Department of Life Sciences, Natural History Museum, Cromwell Road, London, SW7 5BD, UK. t.pennance@nhm.ac.uk.|RVC Department of Pathology and Pathogen Biology, Centre for Emerging, Endemic and Exotic Diseases (CEEED), Royal Veterinary College, University of London, Hertsfordshire, AL97TA, UK. t.pennance@nhm.ac.uk.
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Pennance
Wolfson Wellcome Biomedical Laboratories, Department of Life Sciences, Natural History Museum, Cromwell Road, London, SW7 5BD, UK. a.emery@nhm.ac.uk.
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Emery
Wolfson Wellcome Biomedical Laboratories, Department of Life Sciences, Natural History Museum, Cromwell Road, London, SW7 5BD, UK. f.allan@nhm.ac.uk.
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Allan
Wolfson Wellcome Biomedical Laboratories, Department of Life Sciences, Natural History Museum, Cromwell Road, London, SW7 5BD, UK. a.gouvras@nhm.ac.uk.
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Gouvras
Wolfson Wellcome Biomedical Laboratories, Department of Life Sciences, Natural History Museum, Cromwell Road, London, SW7 5BD, UK. s.knopp@unibas.ch.|Department of Epidemiology and Public Health, Swiss Tropical and Public Health Institute, Socinstrasse 57, 4002, Basel, Switzerland. s.knopp@unibas.ch.|University of Basel, Petersplatz 1, 4003, Basel, Switzerland. s.knopp@unibas.ch.
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Knopp
Réseau International Schistosomoses, Environnement, Aménagement et Lutte (RISEAL-Niger), 333, Avenue des Zarmakoye, B.P. 13724, Niamey, Niger. s.knopp@unibas.ch.
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Garba
Réseau International Schistosomoses, Environnement, Aménagement et Lutte (RISEAL-Niger), 333, Avenue des Zarmakoye, B.P. 13724, Niamey, Niger. amina@risealniger.org.
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AA
Hamidou
Public Health Laboratory - Ivo de Carneri (PHL-IdC), P.O.BOX 122, Wawi, Chake Chake, Pemba, United Republic of Tanzania. kamsharjyy2@yahoo.com.
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Mohammed
Public Health Laboratory - Ivo de Carneri (PHL-IdC), P.O.BOX 122, Wawi, Chake Chake, Pemba, United Republic of Tanzania. shaaliame@yahoo.com.
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Ame
Wolfson Wellcome Biomedical Laboratories, Department of Life Sciences, Natural History Museum, Cromwell Road, London, SW7 5BD, UK. d.rollinson@nhm.ac.uk.
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Rollinson
Department of Infectious Disease Epidemiology, Imperial College Faculty of Medicine (St Mary's Campus), Norfolk Place, London, W2 1PG, UK. jowebster@rvc.ac.uk.|RVC Department of Pathology and Pathogen Biology, Centre for Emerging, Endemic and Exotic Diseases (CEEED), Royal Veterinary College, University of London, Hertsfordshire, AL97TA, UK. jowebster@rvc.ac.uk.
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Webster
D000818:Animals; D003362:Cost-Benefit Analysis; D014644:Genetic Variation; D005828:Genetics, Population; D006801:Humans; D007814:Larva / Q000145:classification / Q000235:genetics; D018895:Microsatellite Repeats; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D009548:Niger; D012548:Schistosoma haematobium / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification; D012553:Schistosomiasis haematobia / Q000469:parasitology; D013636:Tanzania; D013997:Time Factors; D014552:Urinary Tract Infections / Q000469:parasitology