rivulorum has been found infected with the Plasmodium parasite at one locality in Tanzania. vaneedeni has been shown to be susceptible to Plasmodium infection under laboratory conditions, and An. The other species are predominantly zoophilic and exophilic and are thought to have limited, or no, importance as malaria vectors, although An. is an efficient vector of the human malaria parasite and this is reflected by its anthropophilic and endophilic behaviour. The vectorial capacity, biology and behaviour of these species also differ. leesoni have a widespread distribution across sub-Saharan Africa, while An, parensis is found through eastern and southern Africa and An. rivulorum subgroups, respectively, are either morphologically identical or very similar and may occur in sympatry over large parts of their distribution.
funestus subgroup, and Anopheles leesoni and Anopheles rivulorum, belonging to the An. funestus s.s., Anopheles parensis and Anopheles vaneedeni, belonging to the An. The Anopheles funestus group consists of five subgroups of mosquitoes namely Anopheles funestus, Anopheles rivulorum, Anopheles minimus, Anopheles aconitus and Anopheles culicifacies and includes one of the most important African vectors of malaria Anopheles funestus s.s. However, the cost of both the real-time PCR machine and fluorescently labelled probes required is falling and in the future the cost of this assay is likely to become closer to that of standard PCR.
The one disadvantage of the TaqMan assay is the cost of this assay, both in terms of initial capital outlay and running cost per sample, which is higher than AS-PCR. This method is at least as sensitive and specific as the gold standard AS-PCR approach and because it has no requirement for post-PCR processing is simpler and more rapid to run. funestus group using fluorescently-labeled probes with distinct emission and excitation spectra allowing their independent detection in a single reaction. This assay very effectively identified all five members of the An. The TaqMan assay proved to be the most robust of the three protocols tested in this study. The MCA and HRM assays initially gave promising results, but were more sensitive to both DNA quality and quantity and consequently showed a higher rate of incorrect identifications. The TaqMan assay proved to be the most sensitive and specific of the three new assays.
MICROSOFT PROJECT ON MULTIPLEX TRIAL
The sensitivity and specificity of these assays were compared with the widely used AS-PCR in a blind trial using DNA extracted from wild-caught mosquitoes. Protocols for three fluorescence-based assays based on Melt Curve Analysis (MCA), High Resolution Melt (HRM) and TaqMan SNP genotyping were developed to detect and discriminate Anopheles parensis, Anopheles leesoni, Anopheles vaneedeni, Anopheles rivulorum and An. In this study, three new high-throughput 'closed-tube' assays were developed and compared with the previously described AS-PCR technique. One disadvantage of AS-PCR is the requirement for post-PCR processing by gel electrophoresis of PCR products. In the past, this has relied on morphological and cytogenetic methods but these have been largely superseded by a robust allele-specific PCR (AS-PCR). The malaria vector and non-vector species of the Anopheles funestus group are morphologically very similar and accurate identification is required as part of effective control strategies.
0 kommentar(er)
