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A new method for detection of pfmdr1 mutations in Plasmodium falciparum DNA using real-time PCR

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 h...

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Autores principales: Purfield, Anne, Nelson, Amy, Laoboonchai, Anita, Congpuong, Kanungnij, McDaniel, Phillip, Miller, R Scott, Welch, Kathy, Wongsrichanalai, Chansuda, Meshnick, Steven R
Formato: Texto
Lenguaje:English
Publicado: BioMed Central 2004
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC420476/
https://www.ncbi.nlm.nih.gov/pubmed/15132750
http://dx.doi.org/10.1186/1475-2875-3-9
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author Purfield, Anne
Nelson, Amy
Laoboonchai, Anita
Congpuong, Kanungnij
McDaniel, Phillip
Miller, R Scott
Welch, Kathy
Wongsrichanalai, Chansuda
Meshnick, Steven R
author_facet Purfield, Anne
Nelson, Amy
Laoboonchai, Anita
Congpuong, Kanungnij
McDaniel, Phillip
Miller, R Scott
Welch, Kathy
Wongsrichanalai, Chansuda
Meshnick, Steven R
author_sort Purfield, Anne
collection PubMed
description 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.
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spelling pubmed-4204762004-06-11 A new method for detection of pfmdr1 mutations in Plasmodium falciparum DNA using real-time PCR Purfield, Anne Nelson, Amy Laoboonchai, Anita Congpuong, Kanungnij McDaniel, Phillip Miller, R Scott Welch, Kathy Wongsrichanalai, Chansuda Meshnick, Steven R Malar J Methodology 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. BioMed Central 2004-05-07 /pmc/articles/PMC420476/ /pubmed/15132750 http://dx.doi.org/10.1186/1475-2875-3-9 Text en Copyright © 2004 Purfield et al; licensee BioMed Central Ltd. This is an Open Access article: verbatim copying and redistribution of this article are permitted in all media for any purpose, provided this notice is preserved along with the article's original URL.
spellingShingle Methodology
Purfield, Anne
Nelson, Amy
Laoboonchai, Anita
Congpuong, Kanungnij
McDaniel, Phillip
Miller, R Scott
Welch, Kathy
Wongsrichanalai, Chansuda
Meshnick, Steven R
A new method for detection of pfmdr1 mutations in Plasmodium falciparum DNA using real-time PCR
title A new method for detection of pfmdr1 mutations in Plasmodium falciparum DNA using real-time PCR
title_full A new method for detection of pfmdr1 mutations in Plasmodium falciparum DNA using real-time PCR
title_fullStr A new method for detection of pfmdr1 mutations in Plasmodium falciparum DNA using real-time PCR
title_full_unstemmed A new method for detection of pfmdr1 mutations in Plasmodium falciparum DNA using real-time PCR
title_short A new method for detection of pfmdr1 mutations in Plasmodium falciparum DNA using real-time PCR
title_sort new method for detection of pfmdr1 mutations in plasmodium falciparum dna using real-time pcr
topic Methodology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC420476/
https://www.ncbi.nlm.nih.gov/pubmed/15132750
http://dx.doi.org/10.1186/1475-2875-3-9
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