Development of a Real-Time PCR Assay for Detection of Plasmodium falciparum , Plasmodium vivax , and Plasmodium ovale for Routine Clinical Diagnosis
Open accessAbstract
A TaqMan-based real-time PCR qualitative assay for the detection of three species of malaria parasites— Plasmodium falciparum , P. ovale , and P. vivax —was devised and evaluated using 122 whole-blood samples from patients who had traveled to areas where malaria is endemic and who presented with malaria-like symptoms and fever. The assay was compared to conventional microscopy and to an established nested-PCR assay. The specificity of the new assay was confirmed by sequencing the PCR products from all the positive samples and by the lack of cross-reactivity with Toxoplasma gondii and Leishmania infantum DNA. Real-time PCR assay showed a detection limit (analytical sensitivity) of 0.7, 4, and 1.5 parasites/μl for P. falciparum , P. vivax , and P. ovale , respectively. Real-time PCR, like nested PCR, brought to light errors in the species identification by microscopic examination and revealed the presence of mixed infections ( P. falciparum plus P. ovale ). Real-time PCR can yield results within 2 h, does not require post-PCR processing, reduces sample handling, and minimizes the risks of contamination. The assay can therefore be easily implemented in routine diagnostic malaria tests. Future studies are warranted to investigate the clinical value of this technique.
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Real-time quantitative PCR methods have good sensitivity and range that allows monitoring parasitemia levels.
“Real-time quantitative PCR (qPCR) methods have followed, with good sensitivity and range that allows monitoring parasitemia levels [29,30].”
From Diagnosing malaria in pregnancy: An update (2012) · cites this paper as [30]
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- Authors
- Perandin, F., Manca, N., Calderaro, A., Piccolo, G., Galati, L., Ricci, L., Medici, M. C., Arcangeletti, M. C., Snounou, G., Dettori, G., & Chezzi, C.
- Affiliations
- University of Brescia (IT)
- University of Parma (IT)
- Santa Maria Nuova Hospital (IT)
- Centre National de la Recherche Scientifique (FR)
- Corresponding author
- Manca, N.
- Venue
- Journal of Clinical Microbiology
- Publisher
- American Society for Microbiology
- Published
- 2004-03
- Volume
- 42
- Issue
- 3
- Pages
- 1214-1219
- ISSN
- 0095-1137 (ISSN-L)
- 1098-660X
- PMC
- PMC356834
- OpenAlex
- W2088824647
- Cited by
- 354
- References
- 27
- Citations by year
- 2026: 11 · 2025: 13 · 2024: 10 · 2023: 21 · 2022: 20 · 2021: 31 · 2020: 12 · 2019: 11 · 2018: 16 · 2017: 21 · 2016: 12 · 2015: 12 · 2014: 21 · 2013: 27 · 2012: 14
- FWCI
- 16.37
- Type
- journal article
- Language
- en
- Open access
- green
- europepmc.org
- Locations
- Journal of Clinical Microbiology (publishedVersion)
- Europe PMC (PubMed Central) (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Institutional Research Information System (Università degli Studi di Brescia) (submittedVersion)
- Topics
- Health Sciences › Medicine › Public Health, Environmental and Occupational Health › Malaria Research and Control
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Clinical Biochemistry › Bacterial Identification and Susceptibility Testing
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Molecular Biology Techniques and Applications
- Keywords
- Plasmodium falciparum, Plasmodium vivax, Plasmodium ovale, malaria diagnosis, real-time PCR, TaqMan real-time PCR, species identification, mixed malaria infection, diagnostic sensitivity, microscopy comparison, nested PCR comparison, imported malaria, travel-associated malaria
- MeSH terms
- Animals, Base Sequence, Computer Systems, Cross Reactions, Diagnostic Tests, Routine, DNA, Ribosomal, Humans, Malaria, Plasmodium falciparum, Plasmodium vivax, RNA, Ribosomal, 18S, Sensitivity and Specificity, Toxoplasma, Polymerase Chain Reaction, DNA Primers, Leishmania infantum, Genome, Protozoan, Plasmodium ovale
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