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Updated: Sep 2, 2026
Multiplexed Isothermal Amplification Based Diagnostic Platform to Detect Zika, Chikungunya, and Dengue 1
Published on: March 13, 2018
Uracil-DNA Glycosylase-Enhanced Loop-Mediated Isothermal Amplification Assays for Detection of Zoonotic Malaria
Meng Yee Lai1, Mohd Lutfi Abdullah2, Yee Ling Lau1
1Department of Parasitology, Faculty of Medicine, Universiti Malaya, Kuala Lumpur, Malaysia.
Background:
Zoonotic malaria caused by simian Plasmodium species presents increasing diagnostic challenges in endemic regions.
Method:
This study developed and evaluated malachite green-based loop-mediated isothermal amplification (MG-LAMP) assays for detecting five zoonotic malaria parasites (P. knowlesi, P. cynomolgi, P. inui, P. coatneyi, and P. fieldi). The assays enable rapid visual detection through a colorimetric change without requiring specialized equipment. To reduce carryover contamination, deoxyuridine triphosphate (dUTP) and uracil-DNA glycosylase (UDG) were incorporated into the reaction.
Result:
All five UDG MG-LAMP assays demonstrated high analytical sensitivity, with limits of detection of 1 copy/µL for P. cynomolgi, P. coatneyi, and P. fieldi, and 10 copies/µL for P. knowlesi and P. inui. The assays showed 100% specificity, with no cross-reactivity among target species. Malachite green enabled clear visual discrimination between positive and negative reactions.
Conclusion:
A simplified direct blood lysis method requiring less than 20 min was also developed to enable rapid DNA preparation without column-based purification. Overall, the MG-LAMP assays combined with UDG and rapid lysis provide a sensitive, specific, and practical approach for diagnosing zoonotic malaria, particularly in resource-limited endemic settings.
