Single-tube Single-run Multiplex PCR Detects Mixed Samples with Four Species of Plasmodium

To establish a single-tube single-run multiplex PCR technique that can detect single or mixed samples with four species of Plasmodium. Folding primers were designed based on the fast nested PCR. The reaction component concentrations were optimized and the primers were selected based on the annealing...

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Published inZhongguo jishengchongxue yu jishengchongping zazhi Vol. 33; no. 3; p. 200
Main Authors Huang, Yu-ting, She, Dan-ya, Lu, Li-dan, Zhang, Nian-heng, Huang, Tian-yi
Format Journal Article
LanguageChinese
Published China 01.06.2015
Subjects
Coinfection
DNA Primers
Humans
Multiplex Polymerase Chain Reaction
Plasmodium
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Summary:To establish a single-tube single-run multiplex PCR technique that can detect single or mixed samples with four species of Plasmodium. Folding primers were designed based on the fast nested PCR. The reaction component concentrations were optimized and the primers were selected based on the annealing temperature. The established single-tube single-run folding-primer multiplex PCR (FP-PCR) was tested for its sensitivity and specificity to detect single-species and mixed samples with P. vivax, P. falciparum, P. ovale (including P. ovale wallikeri) and/or P. malariae. In all the seven experimental repeats, FP-PCR successfully detected single-species infection for all the four species, with the detection limit reaching or close to 1 parasite/µl blood. For mixed infections with 2-4 species at different densities with the highest being 100 times of the lowest, FP-PCR identified all the species in each combination in 57 out of 84 tests. Further, in 10 dried blood samples on filter paper from healthy subjects, no FP-P
ISSN:1000-7423