Comparison of three molecular methods for the detection and speciation of Plasmodium vivax and Plasmodium falciparum

P Boonma, Peter Raymond Christensen, Rossarin Suwanarusk, Ric Price, B Russell, U Lek-uthai

    Research output: Contribution to journalArticleResearchpeer-review

    Abstract

    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. � 2007 Boonma et al; licensee BioMed Central Ltd.
    Original languageEnglish
    Pages (from-to)124-
    JournalMalaria Journal
    Volume6
    Publication statusPublished - 2007

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    Plasmodium vivax
    Plasmodium falciparum
    Polymerase Chain Reaction
    Malaria
    Multiplex Polymerase Chain Reaction
    Real-Time Polymerase Chain Reaction
    Microscopy
    Sensitivity and Specificity
    Plasmodium
    Thailand
    Freezing
    Cost-Benefit Analysis

    Cite this

    Boonma, P., Christensen, P. R., Suwanarusk, R., Price, R., Russell, B., & Lek-uthai, U. (2007). Comparison of three molecular methods for the detection and speciation of Plasmodium vivax and Plasmodium falciparum. Malaria Journal, 6, 124-.
    Boonma, P ; Christensen, Peter Raymond ; Suwanarusk, Rossarin ; Price, Ric ; Russell, B ; Lek-uthai, U. / Comparison of three molecular methods for the detection and speciation of Plasmodium vivax and Plasmodium falciparum. In: Malaria Journal. 2007 ; Vol. 6. pp. 124-.
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    title = "Comparison of three molecular methods for the detection and speciation of Plasmodium vivax and Plasmodium falciparum",
    abstract = "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. � 2007 Boonma et al; licensee BioMed Central Ltd.",
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    Boonma, P, Christensen, PR, Suwanarusk, R, Price, R, Russell, B & Lek-uthai, U 2007, 'Comparison of three molecular methods for the detection and speciation of Plasmodium vivax and Plasmodium falciparum', Malaria Journal, vol. 6, pp. 124-.

    Comparison of three molecular methods for the detection and speciation of Plasmodium vivax and Plasmodium falciparum. / Boonma, P; Christensen, Peter Raymond; Suwanarusk, Rossarin; Price, Ric; Russell, B; Lek-uthai, U.

    In: Malaria Journal, Vol. 6, 2007, p. 124-.

    Research output: Contribution to journalArticleResearchpeer-review

    TY - JOUR

    T1 - Comparison of three molecular methods for the detection and speciation of Plasmodium vivax and Plasmodium falciparum

    AU - Boonma, P

    AU - Christensen, Peter Raymond

    AU - Suwanarusk, Rossarin

    AU - Price, Ric

    AU - Russell, B

    AU - Lek-uthai, U

    PY - 2007

    Y1 - 2007

    N2 - 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. � 2007 Boonma et al; licensee BioMed Central Ltd.

    AB - 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. � 2007 Boonma et al; licensee BioMed Central Ltd.

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