Development of Cooperative Primer-Based Real-Time PCR Assays for the Detection of Plasmodium malariae and Plasmodium ovale
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Date
2021
Journal Title
Journal ISSN
Volume Title
Publisher
The Journal of Molecular Diagnostics
Abstract
Plasmodium malariae and Plasmodium ovale are increasingly gaining public health attention as the
global transmission of falciparum malaria is decreasing. However, the absence of reliable Plasmodium
species-specific detection tools has hampered accurate diagnosis of these minor Plasmodium species. In
this study, SYBR Greenebased real-time PCR assays were developed for the detection of P. malariae and
P. ovale using cooperative primers that significantly limit the formation and propagation of primersdimers.
Both the P. malariae and P. ovale cooperative primer-based assays had at least 10-fold lower
detection limit compared with the corresponding conventional primer-based assays. More important,
the cooperative primer-based assays were evaluated in a cross-sectional study using 560 samples obtained
from two health facilities in Ghana. The prevalence rates of P. malariae and P. ovale among the
combined study population were 18.6% (104/560) and 5.5% (31/560), respectively. Among the
Plasmodium-positive cases, P. malariae and P. ovale mono-infections were 3.6% (18/499) and 1.0% (5/
499), respectively, with the remaining being co-infections with Plasmodium falciparum. The study
demonstrates the public health importance of including detection tools with lower detection limits in
routine diagnosis and surveillance of nonfalciparum species. This will be necessary for comprehensively
assessing the effectiveness of malaria interventions and control measures aimed toward global malaria
elimination. (
Description
Research Article
Keywords
Plasmodium malariae, Plasmodium ovale, falciparum malaria