title: A sensitive, specific and reproducible real-time polymerase chain reaction method for detection of Plasmodium vivax and Plasmodium falciparum infection in field-collected anophelines.
abstract: We describe a simple method for detection of Plasmodium vivax and Plasmodium falciparum infection in anophelines using a triplex TaqMan real-time polymerase chain reaction (PCR) assay (18S rRNA). We tested the assay on Anopheles darlingi and Anopheles stephensi colony mosquitoes fed with Plasmodium-infected blood meals and in duplicate on field collected An. darlingi. We compared the real-time PCR results of colony-infected and field collected An. darlingi, separately, to a conventional PCR method. We determined that a cytochrome b-PCR method was only 3.33% as sensitive and 93.38% as specific as our real-time PCR assay with field-collected samples. We demonstrate that this assay is sensitive, specific and reproducible.
authors.affiliation: Wadsworth Center, New York State Department of Health, Albany, NY, USA.
authors.forename: Sara A
authors.initials: SA
authors.lastname: Bickersmith
authors.affiliation: Department of Biomedical Sciences, School of Public Health, State University of New York, Albany, NY, USA.
authors.forename: William
authors.initials: W
authors.lastname: Lainhart
authors.affiliation: Division of Infectious Diseases, Department of Medicine, University of California San Diego, La Jolla, CA, USA.
authors.forename: Marta
authors.initials: M
authors.lastname: Moreno
authors.affiliation: Department of Biomedical Sciences, School of Public Health, State University of New York, Albany, NY, USA.
authors.forename: Virginia M
authors.initials: VM
authors.lastname: Chu
authors.affiliation: Division of Infectious Diseases, Department of Medicine, University of California San Diego, La Jolla, CA, USA.
authors.forename: Joseph M
authors.initials: JM
authors.lastname: Vinetz
authors.affiliation: Wadsworth Center, New York State Department of Health, Albany, NY, USA.
authors.forename: Jan E
authors.initials: JE
authors.lastname: Conn
mesh_terms: D000818:Animals; D000852:Anopheles / Q000469:parasitology*; D045303:Cytochromes b / Q000235:genetics; D007303:Insect Vectors / Q000469:parasitology*; D010963:Plasmodium falciparum / Q000235:genetics* / Q000302:isolation & purification; D010966:Plasmodium vivax / Q000235:genetics* / Q000302:isolation & purification; D060888:Real-Time Polymerase Chain Reaction; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D045303:Cytochromes b