Researchers have developed and validated a TaqMan SNP genotyping assay that reliably distinguishes Anopheles messeae from Anopheles daciae, two morphologically indistinguishable members of the Anopheles maculipennis complex that are widely distributed across the Palearctic region. The work, published in Parasites & Vectors, addresses a long-standing obstacle for vector surveillance: because these mosquitoes cannot be told apart by appearance, species-specific monitoring and ecological and epidemiological studies have been hindered.
The assay targets a diagnostic single-nucleotide polymorphism within the Internal Transcribed Spacer 2 (ITS2) region of ribosomal DNA. According to the authors, Mohamed Kader Haidara, Gleb N. Artemov and Igor V. Sharakhov, the method was designed to be rapid, sensitive, specific and suitable for high-throughput screening of field collections.
In validation, the optimized assay was applied to 293 field-collected mosquitoes. It showed high analytical sensitivity, with a detection limit of 0.1 nanograms of genomic DNA, and high specificity, with no false-positive results among the non-target species tested. The assay reliably identified An. messeae, An. daciae and interspecific heterozygotes.
Agreement with an established method was strong. Compared with PCR–RFLP genotyping, the TaqMan assay showed 98 percent overall agreement, corresponding to a Cohen’s kappa of 0.96. The authors also report that the assay is compatible with rapid DNA extraction protocols, which supports its use as a fast and scalable approach for species identification in evolutionary and epidemiological studies.
The authors conclude that the assay provides a rapid and scalable means of differentiating the two species and may facilitate large-scale surveillance of An. messeae and An. daciae populations. Because the article is being shared early as a peer-reviewed, accepted version, some details may be revised in the final Version of Record.
Subject of Research: Development of a TaqMan SNP genotyping assay for molecular identification of the malaria mosquitoes Anopheles messeae and Anopheles daciae
Article Title: Development and validation of a TaqMan SNP genotyping assay for the molecular identification of the malaria mosquitoes Anopheles messeae and Anopheles daciae
Article References: Haidara, M. K., Artemov, G. N., & Sharakhov, I. V. (2026). Development and validation of a TaqMan SNP genotyping assay for the molecular identification of the malaria mosquitoes Anopheles messeae and Anopheles daciae. Parasites & Vectors. https://doi.org/10.1186/s13071-026-07751-0
Image Credits: AI Generated
DOI: 10.1186/s13071-026-07751-0
Keywords: Anopheles messeae, Anopheles daciae, Anopheles maculipennis complex, TaqMan SNP genotyping, ITS2, molecular identification, malaria vectors, vector surveillance, PCR–RFLP, ribosomal DNA, Palearctic mosquitoes, Development
News Source: Drew Townsend. (October 29, 2026). New TaqMan SNP Assay Distinguishes Two Look-Alike Malaria Mosquito Species. Scienmag.
