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Characterization of a Real‐Time PCR Assay for the detection and quantitation of Plasmodium malariae parasites
Author(s) -
Parsons Emily,
Koros Joseph,
Obuya Clifford,
Aching Doty,
Stewart Ann
Publication year - 2011
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.25.1_supplement.540.8
Subject(s) - plasmodium malariae , virology , plasmodium (life cycle) , real time polymerase chain reaction , biology , malaria , plasmodium falciparum , immunology , parasite hosting , genetics , gene , plasmodium vivax , computer science , world wide web
The need for malaria diagnostic tools that are sensitive and specific has propelled the development of real‐time PCR to quantify malaria parasites. The specificity of real‐time PCR is critical for diagnosis of mixed species infections. We have improved the specificity and sensitivity of a real‐time PCR assay for the detection and quantification of Plasmodium malariae , one of four species of malaria parasites, in single and mixed species infections. This real‐time PCR assay amplifies the circumsporozoite (CS) gene of P. malariae . Through primer modification and the use of a sequence‐specific probe, we have increased this assay's sensitivity to detect 1.25 parasites/μl. The limit of quantitation is 3 parasites/μl. We demonstrate that this assay is 100 % specific in single and mixed species infections. We detected no background amplification after 60 amplification cycles and observed no false positives. Compared to expert microscopy as the gold standard, this assay is precise, accurate, and robust. This real‐time PCR assay is suitable to systems requiring high species specificity, including epidemiological studies and diagnoses of returned travelers.