title: Development of PCR-RFLP assay for the discrimination of Plasmodium species and variants of P. vivax (VK210, VK247 and P. vivax-like) in Anopheles mosquitoes.
abstract: The identification of Plasmodium species in Anopheles mosquitoes is an integral component of malaria control programs. We developed a new assay to identify Plasmodium falciparum, Plasmodium malariae, and Plasmodium vivax variants. Specific primers were designed to hybridize to CS gene-specific regions. Polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) were used to distinguish the P. vivax variants VK210, VK247, and P. vivax-like. The new PCR-RFLP assay revealed good agreement when compared with a nested PCR using artificially infected Anopheles mosquitoes. This sensitive PCR-RFLP method can be useful when detection of Plasmodium species and P. vivax variants is required and may be employed to improve the understanding of malaria transmission dynamics by Anopheles species.
authors.affiliation: Universidade de São Paulo, Rua Cristóvão Colombo, São José do Rio Preto, Brazil. gcapatti@hotmail.com
authors.forename: Gustavo Capatti
authors.initials: GC
authors.lastname: Cassiano
authors.forename: Luciane Moreno
authors.initials: LM
authors.lastname: Storti-Melo
authors.forename: Marinete Marins
authors.initials: MM
authors.lastname: Póvoa
authors.forename: Allan Kardec Ribeiro
authors.initials: AK
authors.lastname: Galardo
authors.forename: Andréa Regina Baptista
authors.initials: AR
authors.lastname: Rossit
authors.forename: Ricardo Luiz Dantas
authors.initials: RL
authors.lastname: Machado
mesh_terms: D000818:Animals; D000852:Anopheles / Q000469:parasitology*; D017931:DNA Primers / Q000235:genetics; D010963:Plasmodium falciparum / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D010965:Plasmodium malariae / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D010966:Plasmodium vivax / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D015800:Protozoan Proteins / Q000235:genetics; D012680:Sensitivity and Specificity
chemical_list: D017931:DNA Primers; D015800:Protozoan Proteins; C041595:circumsporozoite protein, Protozoan