Droplet digital polymerase chain reaction (ddPCR) for the detection of Plasmodium knowlesi and Plasmodium vivax

Mahendran, Punitha and Liew, Jonathan Wee Kent and Amir, Amirah and Ching, Xiao-Teng and Lau, Yee-Ling (2020) Droplet digital polymerase chain reaction (ddPCR) for the detection of Plasmodium knowlesi and Plasmodium vivax. Malaria Journal, 19 (1). ISSN 1475-2875, DOI https://doi.org/10.1186/s12936-020-03314-5.

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BackgroundPlasmodium knowlesi and Plasmodium vivax are the predominant Plasmodium species that cause malaria in Malaysia and play a role in asymptomatic malaria disease transmission in Malaysia. The diagnostic tools available to diagnose malaria, such as microscopy and rapid diagnostic test (RDT), are less sensitive at detecting lower parasite density. Droplet digital polymerase chain reaction (ddPCR), which has been shown to have higher sensitivity at diagnosing malaria, allows direct quantification without the need for a standard curve. The aim of this study is to develop and use a duplex ddPCR assay for the detection of P. knowlesi and P. vivax, and compare this method to nested PCR and qPCR.MethodsThe concordance rate, sensitivity and specificity of the duplex ddPCR assay were determined and compared to nested PCR and duplex qPCR.ResultsThe duplex ddPCR assay had higher analytical sensitivity (P. vivax=10 copies/mu L and P. knowlesi=0.01 copies/mu L) compared to qPCR (P. vivax=100 copies/mu L and P. knowlesi=10 copies/mu L). Moreover, the ddPCR assay had acceptable clinical sensitivity (P. vivax=80% and P. knowlesi=90%) and clinical specificity (P. vivax=87.84% and P. knowlesi=81.08%) when compared to nested PCR. Both ddPCR and qPCR detected more double infections in the samples.ConclusionsOverall, the ddPCR assay demonstrated acceptable efficiency in detection of P. knowlesi and P. vivax, and was more sensitive than nested PCR in detecting mixed infections. However, the duplex ddPCR assay still needs optimization to improve the assay's clinical sensitivity and specificity.

Item Type: Article
Uncontrolled Keywords: Droplet digital polymerase chain reaction; Plasmodium knowlesi; Plasmodium vivax; Malaria
Subjects: R Medicine > R Medicine (General)
R Medicine > RA Public aspects of medicine > RA0421 Public health. Hygiene. Preventive Medicine
Divisions: Faculty of Medicine
Faculty of Medicine > Parasitology Deparment
Depositing User: Ms Zaharah Ramly
Date Deposited: 01 Dec 2023 00:19
Last Modified: 01 Dec 2023 00:19
URI: http://eprints.um.edu.my/id/eprint/36554

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