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@article{Bass:2008:10.1186/1475-2875-7-177,
author = {Bass, C and Nikou, D and Blagborough, AM and Vontas, J and Sinden, RE and Williamson, MS and Field, LM},
doi = {10.1186/1475-2875-7-177},
journal = {Malaria Journal},
title = {PCR-based detection of plasmodium in Anopheles mosquitoes: a comparison of a new high-throughput assay with existing methods},
url = {http://dx.doi.org/10.1186/1475-2875-7-177},
volume = {7},
year = {2008}
}

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TY  - JOUR
AB  - Background: Detection of the four malaria-causing Plasmodium species (Plasmodium falciparum, Plasmodium vivax,Plasmodium ovale and Plasmodium malariae) within their mosquito hosts is an essential component of vectorcontrol programmes. Several PCR protocols have been developed for this purpose. Many of these methods, whilesensitive, require multiple PCR reactions to detect and discriminate all four Plasmodium species. In this study anew high-throughput assay was developed and compared with three previously described PCR techniques.Methods: A new assay based on TaqMan SNP genotyping was developed to detect all four Plasmodium speciesand discriminate P. falciparum from P. vivax, P. ovale and P. malariae. The sensitivity and the specificity of the newassay was compared to three alternative PCR approaches and to microscopic dissection of salivary glands in ablind trial of 96 single insect samples that included artificially infected Anopheles stephensi mosquitoes. Theperformance of the assays was then compared using more than 450 field-collected specimens that had beenstored on silica gel, in ethanol or in isopropanol.Results: The TaqMan assay was found to be highly specific when using Plasmodium genomic DNA as template.Tests of analytical sensitivity and the results of the blind trial showed the TaqMan assay to be the most sensitiveof the four methods followed by the 'gold standard' nested PCR approach and the results generated using thesetwo methods were in good concordance. The sensitivity of the other two methods and their agreement with thenested PCR and TaqMan approaches varied considerably. In trials using field collected specimens two of themethods (including the nested protocol) showed a high degree of non-specific amplification when using DNAderived from mosquitoes stored in ethanol or isopropanol. The TaqMan method appeared unaffected when usingthe same samples.Conclusion: This study describes a new high-throughput TaqMan assay that very effectively detects the
AU  - Bass,C
AU  - Nikou,D
AU  - Blagborough,AM
AU  - Vontas,J
AU  - Sinden,RE
AU  - Williamson,MS
AU  - Field,LM
DO  - 10.1186/1475-2875-7-177
PY  - 2008///
SN  - 1475-2875
TI  - PCR-based detection of plasmodium in Anopheles mosquitoes: a comparison of a new high-throughput assay with existing methods
T2  - Malaria Journal
UR  - http://dx.doi.org/10.1186/1475-2875-7-177
UR  - http://hdl.handle.net/10044/1/31222
VL  - 7
ER  -

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