A new method for detection of pfmdr1 mutations in Plasmodium falciparum DNA using real-time PCR -: art. no. 9

被引：41

作者：

Purfield, A

Nelson, A

Laoboonchai, A

Congpuong, K

McDaniel, P

Miller, RS

Welch, K

Wongsrichanalai, C

Meshnick, SR ^{[1
]}

机构：

[1] Univ N Carolina, Sch Med, Dept Microbiol & Immunol, Chapel Hill, NC 27599 USA

[2] Univ N Carolina, Sch Publ Hlth, Dept Epidemiol, Chapel Hill, NC USA

[3] Armed Forces Res Inst Med Sci, Dept Immunol, Bangkok 10400, Thailand

[4] Minist Publ Hlth, Dept Dis Control, Bur Vector Dis, Bangkok, Thailand

[5] Kwai River Christian Hosp, Sangkhlaburi, Kanchanaburi, Thailand

[6] USN, Med Res Unit, Jakarta, Indonesia

来源：

MALARIA JOURNAL | 2004年 / 3卷 / 1期

关键词：

D O I：

10.1186/1475-2875-3-9

中图分类号：

R51 [传染病];

学科分类号：

100401 ;

摘要：

Background: Surveillance for drug-resistant Plasmodium falciparum should be a component of malaria control programmes. Real-time PCR methods for the detection of parasite single-nucleotide polymorphisms ( SNPs) and gene amplification could be useful survellance tools. Methods: A real-time PCR assay has been developed that identifies single nucleotide polymorphisms (SNPs) at amino acids 86, 184, 1034 and 1042 in the P. falciparum multi-drug resistant (pfmdr 1) gene that may be associated with anti-malarial drug resistance. Results: This assay has a sensitivity and specificity of 94% and 100% when compared to traditional PCR methods for genotyping. Only 54 of 68 (79%) paired pre- and post-culture DNA samples were concordant at all four loci. Conclusion: Real-time PCR is a sensitive and specific method to detect SNP's in pfmdr 1. Genotypes of parasites after in vitro culture may not reflect that seen in vivo.

引用

页数：6

共 17 条

[1] Efficient priming of PCR with short oligonucleotides conjugated to a minor groove binder [J].