1921
Volume 72, Issue 2
  • ISSN: 0002-9637
  • E-ISSN: 1476-1645

Abstract

Single nucleotide polymorphisms (SNPs) in the dihydrofolate reductase (), and dihydropteroate synthetase (), and chloroquine resistance transporter () genes are used as molecular markers of resistance to sulfadoxine/pyrimethamine and chloroquine. However, to be a practical tool in the surveillance of drug resistance, simpler methods for high-throughput haplotyping are warranted. Here we describe a quick and simple technique that detects , , and SNPs using polymerase chain reaction (PCR)– and enzyme-linked immunosorbent assay (ELISA)–based technology. Biotinylated PCR products of , , or were captured on streptavidin-coated microtiter plates and sequence-specific oligonucleotide probes (SSOPs) were hybridized with the PCR products. A stringent washing procedure enabled detection of remaining bound SSOPs and distinguished between the SNPs of , , and with high specificity. The SSOP-ELISA compared well with a standard PCR–restriction fragment length polymorphism procedure, and gave identical positive results in more than 90% of the slide-positive samples tested. The SSOP-ELISA of all , , or SNPs on 88 samples can be performed in a single day and provides quick and reproducible results. The system can potentially be modified to detect SNPs in other genes.

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2005-02-01
2017-07-28
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  • Received : 14 Jul 2004
  • Accepted : 09 Oct 2004

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