Molecular Marker Confi rmation for Member of Anopheles barbirostris Van Der Wulp 1884 in Different Localities  

Oleh:

Satoto, Tri Baskoro Tunggul

Jenis: Article from Journal - ilmiah nasional - terakreditasi DIKTI
Dalam koleksi: Indonesian Journal of Biotechnology vol. 17 no. 2 (Dec. 2012), page 163-168.
Topik: Anopheles barbirostris; ITS2; PCR; Specifi c primer diagnostic
Fulltext: 144_Ros.pdf (184.86KB)

Isi artikel

Vector and non-vector forms of Anopheles barbirostris have been recognized in Indonesia. However, because of their similarity in morphology, they were considered to be a single species. This information has led to the hypothesis that Anopheles barbirostris is a complex of species, which are morphologically indistinguishable from each other by ordinary methods. Objectives of the research was to identify the member of Anopheles barbirostris by PCR Assay. Samples were taken from two localities in Java, two in Sulawesi, two in Flores Indonesia, one from Thailand, one from China. The study was to develop a PCR-based technique of rDNA ITS2 region. Results showed that there are at least four species within the Anopheles barbirostris population studied, namely Anopheles barbirostris species DW, DX, DY and DZ. The length of the sequence amplifi ed for species W, species X, species Y, and species Z were 339bps, 247bps, 165bps. and 157bps, respectively. Verifi cation of the method was carried out with 270 mosquitoes from eight different fi eld-collection sites using various sampling methods. Samples collected from Singaraja-Flores were identifi ed as species W and X. All specimens collected from human bite outdoors were identifi ed as species X; this species showed to be predominant among indoor light trap, indoor human bite and indoor resting collections Samples from Reo-Flores were identifi ed as species W and X. All specimens from Manado and Palopo in Sulawesi were identifi ed as species Z. Similarly only species Y was found in samples from Thailand, while specimens from Salaman and Jambu in Java were identifi ed as species W or species X. These species-specifi c molecular markers for the Anopheles barbirostris, complex appear to be reliable over a wide geographical area. However, larger number of samples is still needed from throughout the range of this species.

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