Da. Oliveira et al., POLYMERASE CHAIN-REACTION AND A LIQUID-PHASE, NONISOTOPIC HYBRIDIZATION FOR SPECIES-SPECIFIC AND SENSITIVE DETECTION OF MALARIA INFECTION, The American journal of tropical medicine and hygiene, 52(2), 1995, pp. 139-144
Citations number
28
Categorie Soggetti
Public, Environmental & Occupation Heath","Tropical Medicine
In the present study, we describe a polymerase chain reaction (PCR)-ba
sed enzyme-linked immunosorbent assay for the detection of malaria inf
ection. The target region of the 18S ribosomal DNA is amplified by a P
CR using an 18S rRNA, genus-specific, biotinylated (5') and an unlabel
ed primer (3') pair. The detection probes are digoxigenin-labeled DNA
oligonucleotides derived from species-specific rRNA sequences. The amp
lified fragments are allowed to hybridize with the species-specific, d
igoxigenin-labeled oligonucleotide probes. The oligo/DNA complex is al
lowed to bind onto streptavidin-coated microtiter plates, followed by
incubation with a peroxidase-streptavidin conjugate and a colorimetric
-peroxidase substrate. The resulting test demonstrated specificity for
the four human Plasmodium species, and was able to detect a level of
parasitemia of at least 0.0001% in a laboratory-induced P. falciparum
infection in monkeys, This liquid hybridization assay is sensitive, sp
ecific, simple, and reliable, with wide applicability in epidemiologic
studies, accurate detection of mixed infections, detection of low-lev
el parasitemia, and evaluation of chemotherapy and vaccine efficacy.