Demonstration by the polymerase chain reaction of mixed Plasmodium falciparum and P. vivax infections undetected by conventional microscopy
Abstract
Mixed malaria infections (Plasmodium falciparum and P. vivax) are suspected to occur at a greater frequency than is detected by conventional light microscopy. To determine this frequency we carried out a prospective 'blinded' comparison of diagnosis by conventional light microscopy and enzymatic amplification of the circumsporozoite gene extracted from dried spotted blood samples. Patients were previously healthy, active duty Thai soldiers assigned to a malaria risk area presenting with malaria. Microscopy (oil immersion objective at 1000 x magnification) involved examination of Giemsa-stained thick and thin blood films by an experienced microscopist. Whole blood samples (25 microliters) dried on filter paper were used for species-specific parasite deoxyribonucleic acid (DNA) amplification by the polymerase chain reaction (PCR) and hybridization with radiolabelled P. falciparum and P. vivax probes. Of 137 consecutive cases of malaria studied, 9% (3/32) of microscopically diagnosed P. falciparum infections and 5% (5/104) of microscopically diagnosed P. vivax infections were found to be mixed by the PCR/DNA probe systems, while 1 case was diagnosed as mixed by both microscopy and PCR. The possibility that malaria patients may have undetected mixed infections should be kept in mind because of the specific therapy required both for P. falciparum and for radical cure of P. vivax.
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Molecular detection for Plasmodium diagnosis using PCR has resulted in increased sensitivity and species discrimination compared to either microscopic or immunochromatographic diagnosis of malaria.
“Molecular detection for Plasmodium diagnosis using PCR has resulted in increased sensitivity and species discrimination compared to either microscopic or immunochromatographic diagnosis of malaria (2, 4, 12, 14, 22, 23, 25, 27).”
From Real-Time PCR for Detection and Identification of <i>Plasmodium</i> spp (2005) · cites this paper as [4]
Several studies reported posttreatment detection of Plasmodium DNA up to 4 days after microscopic detection.
“Several studies reported the posttreatment detection of Plasmodium DNA up to 4 days after microscopic detection (4, 25)”
From Real-Time PCR for Detection and Identification of <i>Plasmodium</i> spp (2005) · cites this paper as [4]
Most published PCR assays are gel based with or without subsequent probe hybridizations.
“However, most published PCR assays are gel based with (4, 5, 15) or without (2, 3, 9, 23, 25-27, 29, 31, 35, 36) subsequent probe hybridizations”
From Real-Time PCR for Detection and Identification of <i>Plasmodium</i> spp (2005) · cites this paper as [4]
A drawback common to all DNA-based malaria assays is the real possibility of detecting persistent DNA in the bloodstream after the infection has subsided.
“A drawback common to all DNA-based malaria assays is the real possibility of detecting persistent DNA in the bloodstream after the infection has subsided.”
From Real-Time PCR for Detection and Identification of <i>Plasmodium</i> spp (2005) · cites this paper as [4]
Many published studies show improved sensitivity and specificity of PCR-based assays over microscopic or immunochromatographic diagnosis of malaria.
“Although there are many published studies showing the improved sensitivity and specificity of PCR-based assays over microscopic or immunochromatographic diagnosis of malaria (2, 4, 12, 14, 22, 23, 25, 27), only a handful take advantage of the even more sensitive and time-efficient real-time PCR technologies.”
From Real-Time PCR for Detection and Identification of <i>Plasmodium</i> spp (2005) · cites this paper as [4]
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Okell, L. C., Ghani, A. C., Lyons, E., & Drakeley, C. J. (2009) · The Journal of Infectious Diseases · doi:10.1086/644781 · Cited by 532 · see citing works
Real-Time PCR for Detection and Identification of <i>Plasmodium</i> spp
Mangold, K. A., Manson, R. U., Koay, E. S. C., Stephens, L., Regner, M., Thomson, R. B., Peterson, L. R., & Kaul, K. L. (2005) · Journal of Clinical Microbiology · doi:10.1128/jcm.43.5.2435-2440.2005 · Cited by 278 · see citing works
Wongsrichanalai, C., Chuanak, N., Tulyayon, S., Thanoosingha, N., Laoboonchai, A., Thimasarn, K., Brewer, T. G., & Heppner, D. G. (1999) · Acta Tropica · doi:10.1016/s0001-706x(99)00040-6 · Cited by 57 · see citing works
Osman, M. M. M., Nour, B. Y. M., Sedig, M. F., De Bes, L., Babikir, A. M., Mohamedani, A. A., & Mens, P. F. (2010) · Tropical Medicine & International Health · doi:10.1111/j.1365-3156.2010.02659.x · Cited by 40 · see citing works
- Authors
- Brown, A. E., Kain, K. C., Pipithkul, J., & Webster, H. K.
- Affiliations
- Armed Forces Research Institute of Medical Science (TH)
- Walter Reed Army Institute of Research (US)
- Venue
- Transactions of the Royal Society of Tropical Medicine and Hygiene
- Publisher
- Oxford University Press (OUP)
- Published
- 1992-11
- Volume
- 86
- Issue
- 6
- Pages
- 609-612
- ISSN
- 0035-9203 (ISSN-L)
- OpenAlex
- W2031653118
- Cited by
- 136
- References
- 0
- Citations by year
- 2026: 1 · 2023: 1 · 2022: 3 · 2021: 4 · 2020: 2 · 2018: 2 · 2017: 2 · 2016: 5 · 2015: 4 · 2014: 5 · 2013: 4 · 2012: 5
- FWCI
- 4.17
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Transactions of the Royal Society of Tropical Medicine and Hygiene (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Health Sciences › Medicine › Public Health, Environmental and Occupational Health › Malaria Research and Control
- Life Sciences › Immunology and Microbiology › Parasitology › Bird parasitology and diseases
- Life Sciences › Immunology and Microbiology › Parasitology › Vector-borne infectious diseases
- Keywords
- mixed malaria infection, Plasmodium falciparum, Plasmodium vivax, PCR diagnosis, microscopic diagnosis, underdiagnosis, dried blood spots, species-specific DNA probes, radical cure
- MeSH terms
- Amino Acid Sequence, Humans, Molecular Sequence Data, Prospective Studies, Oligonucleotide Probes, Protozoan Proteins, DNA, Protozoan, Polymerase Chain Reaction, Malaria, Falciparum, Malaria, Vivax
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