A visualized hybrid PCR-LAMP assay for the detection of human Plasmodium species

Lai, Meng Yee and Ponnampalavanar, Sasheela Sri La Sri and Omar, Sharifah Faridah Syed and Lau, Yee Ling (2024) A visualized hybrid PCR-LAMP assay for the detection of human Plasmodium species. Acta Tropica, 251. p. 107120. ISSN 0001-706X, DOI https://doi.org/10.1016/j.actatropica.2024.107120.

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Official URL: https://doi.org/10.1016/j.actatropica.2024.107120

Abstract

Combining the advantages of PCR and LAMP, we described a new technique, namely PCR-LAMP, for malaria diagnosis. The whole process of DNA amplification can be completed in 35 min. This hybrid amplification technique markedly improved the sensitivity of detection compared to the classic single PCR or LAMP assay alone. PCR-LAMP assay had a detection limit of 1 copy/mu L for P. knowlesi and P. ovale, 0.1 copy/mu L for P. vivax, P. falciparum and P. malariae, respectively. To facilitate the endpoint detection, xylenol orange was added. Positive samples were indicated in orange while negative reactions were violet. The inclusion of xylenol orange into the LAMP reaction mix significantly reduces the post -amplification workload. Without relying on the use of specific instruments, the color changes of the amplicons could be visualized directly through the naked eye. In conclusion, PCR-LAMP poses the potential to be developed as a new malaria molecular diagnosis tool.

Item Type: Article
Funders: Long Term Research Grant Scheme of the Ministry of Higher Education, Malaysia (LRGS/1/2018/UM/01/1/4)
Uncontrolled Keywords: LAMP; PCR; Isothermal; Xylenol orange; Molecular detection; P. knowlesi
Subjects: Q Science > QR Microbiology
R Medicine > R Medicine (General)
Divisions: Faculty of Medicine > Medicine Department
Faculty of Medicine > Parasitology Deparment
Depositing User: Ms. Juhaida Abd Rahim
Date Deposited: 12 Nov 2024 02:50
Last Modified: 12 Nov 2024 02:50
URI: http://eprints.um.edu.my/id/eprint/45787

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