A single-round multiplex PCR assay for the identification of Anopheles minimus related species infected with Plasmodium falciparum and Plasmodium vivax
| Autor: | Usa Lek-Uthai, Paiboon Eamkum, Onanong Kritpetcharat, Panutas Kritpetcharat, Jureerat Daduang, Lertchai Charerntanyarak, Sungsit Sungvornyothin |
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| Rok vydání: | 2014 |
| Předmět: |
Base Sequence
biology Molecular Sequence Data Plasmodium falciparum Plasmodium vivax Anopheles aconitus Anopheles varuna Thailand biology.organism_classification Anopheles minimus Polymerase Chain Reaction Sensitivity and Specificity Virology Plasmodium Anopheles harrisoni Infectious Diseases Species Specificity Anopheles parasitic diseases Multiplex polymerase chain reaction Animals DNA Intergenic Parasitology |
| Zdroj: | Parasitology International. 63:442-449 |
| ISSN: | 1383-5769 |
| DOI: | 10.1016/j.parint.2013.11.001 |
| Popis: | article i nfo 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 com- bined with three Plasmodium primers for the detection of P. falciparum and P. vivax in vectors. The four species of anopheline vectors and twoPlasmodiumspecies, P. falciparum and P. vivax, could be identified by the combina- tion 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. |
| Databáze: | OpenAIRE |
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