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...

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Autores principales: Boonma, Prapaporn, Christensen, Peter R, Suwanarusk, Rossarin, Price, Ric N, Russell, Bruce, Lek-Uthai, Usa
Formato: Texto
Lenguaje:English
Publicado: BioMed Central 2007
Materias:
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.
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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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