Vector borne and zoonotic diseases (Larchmont, N.Y.) · 2026
Uracil-DNA Glycosylase-Enhanced Loop-Mediated Isothermal Amplification Assays for Detection of Zoonotic Malaria Plasmodium Parasites.
СтатьяНазвание и аннотация: Перевод в очереди
Обработка статьи · этапы, ошибки и управление
Загрузка состояния…
Ошибки и ограничения
История · было → стало
Состояние обновляется каждые 5 секунд, пока блок открыт. Процент показывается только там, где известен объём работы. Пауза останавливает продолжение этапа; текущий запрос к источнику может завершиться.
Lai Meng Yee, Abdullah Mohd Lutfi, Lau Yee Ling
Аннотация
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.
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.