title: Use of the polymerase chain reaction to directly detect malaria parasites in blood samples from the Venezuelan Amazon.
abstract: We have examined the reproducibility, sensitivity, and specificity of detecting Plasmodium falciparum using the polymerase chain reaction (PCR) and the species-specific probe pPF14 under field conditions in the Venezuelan Amazon. Up to eight samples were field collected from each of 48 consenting Amerindians presenting with symptoms of malaria. Sample processing and analysis was performed at the Centro Amazonico para la Investigacion y Control de Enfermedades Tropicales Simon Bolivar. A total of 229 samples from 48 patients were analyzed by PCR methods using four different P. falciparum-specific probes. One P. vivax-specific probe and by conventional microscopy. Samples in which results from PCR and microscopy differed were reanalyzed at a higher sensitivity by microscopy. Results suggest that microscopy-negative, PCR-positive samples are true positives, and that microscopy-positive and PCR-negative samples are true negatives. The sensitivity of the DNA probe/PCR method was 78% and its specificity was 97%. The positive predictive value of the PCR method was 88%, and the negative predictive value was 95%. Through the analysis of multiple blood samples from each individual, the DNA probe/PCR methodology was found to have an inherent reproducibility that was highly statistically significant.
authors.affiliation: Department of Population and International Health, Harvard School of Public Health, Boston, Massachusetts.
authors.forename: K F
authors.initials: KF
authors.lastname: Laserson
authors.forename: I
authors.initials: I
authors.lastname: Petralanda
authors.forename: D M
authors.initials: DM
authors.lastname: Hamlin
authors.forename: R
authors.initials: R
authors.lastname: Almera
authors.forename: M
authors.initials: M
authors.lastname: Fuentes
authors.forename: A
authors.initials: A
authors.lastname: Carrasquel
authors.forename: R H
authors.initials: RH
authors.lastname: Barker
mesh_terms: D000818:Animals; D001483:Base Sequence; D017931:DNA Primers / Q000737:chemistry; D015342:DNA Probes; D016054:DNA, Protozoan / Q000097:blood* / Q000737:chemistry; D006801:Humans; D007199:Indians, South American; D016778:Malaria, Falciparum / Q000097:blood / Q000175:diagnosis*; D016780:Malaria, Vivax / Q000097:blood / Q000175:diagnosis; D008969:Molecular Sequence Data; D010963:Plasmodium falciparum / Q000235:genetics / Q000302:isolation & purification*; D010966:Plasmodium vivax / Q000235:genetics / Q000302:isolation & purification; D016133:Polymerase Chain Reaction; D011237:Predictive Value of Tests; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013045:Species Specificity; D014687:Venezuela
chemical_list: D017931:DNA Primers; D015342:DNA Probes; D016054:DNA, Protozoan