Publication: A single-round multiplex PCR assay for the identification of Anopheles minimus related species infected with Plasmodium falciparum and Plasmodium vivax
4
Issued Date
2014-04-01
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ISSN
18730329
13835769
13835769
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2-s2.0-84894418927
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Mahidol University
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SCOPUS
Bibliographic Citation
Parasitology International. Vol.63, No.2 (2014), 442-449
Suggested Citation
Paiboon Eamkum, Sungsit Sungvornyothin, Onanong Kritpetcharat, Jureerat Daduang, Usa Lek-Uthai, Lertchai Charerntanyarak, Panutas Kritpetcharat A single-round multiplex PCR assay for the identification of Anopheles minimus related species infected with Plasmodium falciparum and Plasmodium vivax. Parasitology International. Vol.63, No.2 (2014), 442-449. doi:10.1016/j.parint.2013.11.001 Retrieved from: https://repository.li.mahidol.ac.th/handle/123456789/33975
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Title
A single-round multiplex PCR assay for the identification of Anopheles minimus related species infected with Plasmodium falciparum and Plasmodium vivax
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Abstract
This study aimed to develop a single-round multiplex PCR method for the identification of Anopheles minimus complex (An. minimus and Anopheles harrisoni) and Anopheles aconitus subgroup (An. aconitus and Anopheles varuna), and for the simultaneous detection of Plasmodium falciparum and Plasmodium vivax in these vectors. Five primers were created for a single-round multiplex PCR assay to identify four anopheline mosquitoes combined with three Plasmodium primers for the detection of P. falciparum and P. vivax in vectors. The four species of anopheline vectors and two Plasmodium species, P. falciparum and P. vivax, could be identified by the combination of eight primers in the single-round multiplex PCR assay. The amplified species-specific products were 380. bp for An. minimus, 180. bp for An. harrisoni, 150. bp for An. aconitus, 310. bp for An. varuna, 276. bp for P. falciparum, and 300. bp for P. vivax. The sensitivities were 0.5. pg/μl (25. sporozoites/μl) for P. falciparum DNA and between 0.5 and 5. pg/μl (25-250. sporozoites/μl) for P. vivax DNA. Furthermore, this developed method could be used to identify field caught An. minimus complex, An. aconitus subgroup from Thailand and Lao PDR. Also, it was successfully used to identify the species An. minimus, An. harrisoni, An. aconitus and An. varuna and to detect and identify P. falciparum and P. vivax in caught anopheline mosquitoes. The sensitivity of this method was high for simultaneous detection of P. falciparum and P. vivax in anopheline mosquitoes. © 2013 Elsevier Ireland Ltd.