Abstract
BackgroundRecent studies presented two clades (clades I and II) within the major malaria vector, Anopheles funestus s.s. on the mitochondrial DNA. We describe a hydrolysis probe analysis (Taqman assay) method for the rapid identification of these two clades.FindingsA total of 53 An. funestus s.s. from Malawi and Mozambique were tested for detection of clade types using the hydrolysis probe analysis. Results were compared to DNA sequence analysis to verify the accuracy of the probes Taqman assay for this vector species. Analysis using the hydrolysis probe revealed that there were 21 individuals from Malawi and 13 individuals from Mozambique for clade I, and 19 individuals from Mozambique for clade II. The results were consistent with the results of DNA sequences. A field sample from northern Zambia revealed the presence of both clade types.ConclusionA diagnostic method using the hydrolysis probe analysis was developed to identify clade types within An. funestus s.s. This assay will be useful for screening clade types of field-collected An. funestus specimens accurately and efficiently in malaria vector research and control studies.
Highlights
Recent studies presented two clades within the major malaria vector, Anopheles funestus s.s. on the mitochondrial DNA
A diagnostic method using the hydrolysis probe analysis was developed to identify clade types within An. funestus s.s. This assay will be useful for screening clade types of field-collected An. funestus specimens accurately and efficiently in malaria vector research and control studies
We report on the use of a hydrolysis probe assay (Taqman assay) that is able to accurately and rapidly identify the two clade types of An. funestus s.s
Summary
The application of the assay described here is expected to greatly improve the efficiency of screening largescale field-collected samples of An. funestus s.s. for clades I and II. This assay could help elucidate the role that each clade is playing in malaria transmission and whether there is any difference in insecticide resistance frequencies. Competing interests The authors declare no competing interests. Authors’ contributions KSC designed the study, developed the new hydrolysis probe analysis and drafted the manuscript. LLK and MC assisted with analysis of the data and helped draft the manuscript. All authors read and approved the final manuscript
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