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J. Clin. Microbiol. doi:10.1128/JCM.00081-08
Copyright (c) 2008, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

Multiplex-PCR and oligonucleotide microarray for detection of single nucleotide polymorphisms associated with drug resistance in Plasmodium falciparum

National Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention, WHO Collaborating Center for Malaria, Schistosomiasis and Filariasis, 207 Rui Jin Er Road, Shanghai 200025, China; National Engineering Center for Biochip at Shanghai, 151 Li Bing Road, Pudong, Shanghai 201203, China

^* To whom correspondence should be addressed. Email: ipd_sh{at}163.com.

	Abstract

Drug resistance in Plasmodium falciparum is a serious public health threat in the endemic countries. Since resistance has been associated with specific single nucleotide polymorphisms (SNPs) in the parasite genes, molecular markers are becoming useful surrogates for monitoring the emergence and dispersion of drug resistance. In this study, a multiplex-PCR (mPCR) and oligonucleotide microarray method was developed for detection of these SNPs in genes encoding chloroquine resistance transporter (pfcrt), multi-drug resistance 1 (pfmdr1), dihydrofolate reductase (pfdhfr), dihydropteroate synthetase (pfdhps) and adenosine triphosphatase-6 (pfATPase6) of P. falciparum. The results show that DNA microarray technology combined with multiplex PCR is a promising and time-saving tool supporting conventional detection methods, allowing sensitive, accurate, simultaneous analysis of the SNPs associated with drug resistance in P. falciparum.