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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...
Autores principales: | , , , , , , , , |
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Formato: | Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2004
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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. |
format | Text |
id | pubmed-420476 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2004 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
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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