Comparison of three molecular methods for the detection and speciation of Plasmodium vivax and Plasmodium falciparum
BACKGROUND: Accurate diagnosis of Plasmodium spp. is essential for the rational treatment of malaria. Despite its many disadvantages, microscopic examination of blood smears remains the current "gold standard" for malaria detection and speciation. PCR assays offer an alternative to microsc...
Autores principales: | , , , , , |
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Formato: | Texto |
Lenguaje: | English |
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BioMed Central
2007
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2020467/ https://www.ncbi.nlm.nih.gov/pubmed/17868467 http://dx.doi.org/10.1186/1475-2875-6-124 |
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author | Boonma, Prapaporn Christensen, Peter R Suwanarusk, Rossarin Price, Ric N Russell, Bruce Lek-Uthai, Usa |
author_facet | Boonma, Prapaporn Christensen, Peter R Suwanarusk, Rossarin Price, Ric N Russell, Bruce Lek-Uthai, Usa |
author_sort | Boonma, Prapaporn |
collection | PubMed |
description | BACKGROUND: Accurate diagnosis of Plasmodium spp. is essential for the rational treatment of malaria. Despite its many disadvantages, microscopic examination of blood smears remains the current "gold standard" for malaria detection and speciation. PCR assays offer an alternative to microscopy which has been shown to have superior sensitivity and specificity. Unfortunately few comparative studies have been done on the various molecular based speciation methods. METHODS: The sensitivity, specificity and cost effectiveness of three molecular techniques were compared for the detection and speciation of Plasmodium falciparum and Plasmodium vivax from dried blood spots collected from 136 patients in western Thailand. The results from the three molecular speciation techniques (nested PCR, multiplex PCR, and real-time PCR) were used to develop a molecular consensus (two or more identical PCR results) as an alternative gold standard. RESULTS: According to the molecular consensus, 9.6% (13/136) of microscopic diagnoses yielded false negative results. Multiplex PCR failed to detect P. vivax in three mixed isolates, and the nested PCR gave a false positive P. falciparum result in one case. Although the real-time PCR melting curve analysis was the most expensive method, it was 100% sensitive and specific and least time consuming of the three molecular techniques investigated. CONCLUSION: Although microscopy remains the most appropriate method for clinical diagnosis in a field setting, its use as a gold standard may result in apparent false positive results by superior techniques. Future studies should consider using more than one established molecular methods as a new gold standard to assess novel malaria diagnostic kits and PCR assays. |
format | Text |
id | pubmed-2020467 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2007 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-20204672007-10-13 Comparison of three molecular methods for the detection and speciation of Plasmodium vivax and Plasmodium falciparum Boonma, Prapaporn Christensen, Peter R Suwanarusk, Rossarin Price, Ric N Russell, Bruce Lek-Uthai, Usa Malar J Methodology BACKGROUND: Accurate diagnosis of Plasmodium spp. is essential for the rational treatment of malaria. Despite its many disadvantages, microscopic examination of blood smears remains the current "gold standard" for malaria detection and speciation. PCR assays offer an alternative to microscopy which has been shown to have superior sensitivity and specificity. Unfortunately few comparative studies have been done on the various molecular based speciation methods. METHODS: The sensitivity, specificity and cost effectiveness of three molecular techniques were compared for the detection and speciation of Plasmodium falciparum and Plasmodium vivax from dried blood spots collected from 136 patients in western Thailand. The results from the three molecular speciation techniques (nested PCR, multiplex PCR, and real-time PCR) were used to develop a molecular consensus (two or more identical PCR results) as an alternative gold standard. RESULTS: According to the molecular consensus, 9.6% (13/136) of microscopic diagnoses yielded false negative results. Multiplex PCR failed to detect P. vivax in three mixed isolates, and the nested PCR gave a false positive P. falciparum result in one case. Although the real-time PCR melting curve analysis was the most expensive method, it was 100% sensitive and specific and least time consuming of the three molecular techniques investigated. CONCLUSION: Although microscopy remains the most appropriate method for clinical diagnosis in a field setting, its use as a gold standard may result in apparent false positive results by superior techniques. Future studies should consider using more than one established molecular methods as a new gold standard to assess novel malaria diagnostic kits and PCR assays. BioMed Central 2007-09-15 /pmc/articles/PMC2020467/ /pubmed/17868467 http://dx.doi.org/10.1186/1475-2875-6-124 Text en Copyright © 2007 Boonma et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( (http://creativecommons.org/licenses/by/2.0) ), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Methodology Boonma, Prapaporn Christensen, Peter R Suwanarusk, Rossarin Price, Ric N Russell, Bruce Lek-Uthai, Usa Comparison of three molecular methods for the detection and speciation of Plasmodium vivax and Plasmodium falciparum |
title | Comparison of three molecular methods for the detection and speciation of Plasmodium vivax and Plasmodium falciparum |
title_full | Comparison of three molecular methods for the detection and speciation of Plasmodium vivax and Plasmodium falciparum |
title_fullStr | Comparison of three molecular methods for the detection and speciation of Plasmodium vivax and Plasmodium falciparum |
title_full_unstemmed | Comparison of three molecular methods for the detection and speciation of Plasmodium vivax and Plasmodium falciparum |
title_short | Comparison of three molecular methods for the detection and speciation of Plasmodium vivax and Plasmodium falciparum |
title_sort | comparison of three molecular methods for the detection and speciation of plasmodium vivax and plasmodium falciparum |
topic | Methodology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2020467/ https://www.ncbi.nlm.nih.gov/pubmed/17868467 http://dx.doi.org/10.1186/1475-2875-6-124 |
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