End-point detection of loop-mediated isothermal amplification (LAMP) on malaria by direct observation with colorimetric dyes.

Autor: Zen LPY; Department of Parasitology, Faculty of Medicine, Universiti Malaya, 50603, Kuala Lumpur, Malaysia., Lai MY; Department of Parasitology, Faculty of Medicine, Universiti Malaya, 50603, Kuala Lumpur, Malaysia., Izzati Binti Rozlan S; Department of Parasitology, Faculty of Medicine, Universiti Malaya, 50603, Kuala Lumpur, Malaysia., Abdul Hamid MH; Vector Borne Disease Sector, Ministry of Health, Putrajaya, Malaysia., Jelip J; Vector Borne Disease Sector, Ministry of Health, Putrajaya, Malaysia., Nani Mudin R; Vector Borne Disease Sector, Ministry of Health, Putrajaya, Malaysia., Lau YL; Department of Parasitology, Faculty of Medicine, Universiti Malaya, 50603, Kuala Lumpur, Malaysia. Electronic address: lauyeeling@um.edu.my.
Jazyk: angličtina
Zdroj: Experimental parasitology [Exp Parasitol] 2022 Aug; Vol. 239, pp. 108310. Date of Electronic Publication: 2022 Jun 17.
DOI: 10.1016/j.exppara.2022.108310
Abstrakt: In order to ascertain the results of the LAMP technique, different end-point detection methods can be employed. However, these methods require sophisticated equipment. To simplify current end-point detection methods for the diagnosis of malaria, we propose the incorporation of colorimetric dyes: malachite green (MG), phenol red (PR), and xylenol orange (XO) in the LAMP assay. To evaluate the optimum concentration of dyes, 5 different concentrations (50 μM, 75 μM, 100 μM, 125 μM, and 150 μM) were used with buffer pH 8.5 and pH 8.8, respectively. The results showed that 125 μM of MG at pH 8.8 produced the most obvious colour change. A total of 71 clinical blood samples of Plasmodium knowlesi, Plasmodium malariae, Plasmodium vivax, Plasmodium falciparum, and healthy donors were tested using MG-LAMP. It showed 100% sensitivity and specificity. The simplicity and affordability of this method make it ideal to be used as an end-point detection method for malaria diagnosis in resource limited settings.
(Copyright © 2022 Elsevier Inc. All rights reserved.)
Databáze: MEDLINE
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