Lai Meng-Yee, Abdul Hamid Mohd Hafizi, Jelip Jenarun, Mudin Rose Nani, Lau Yee-Ling
Department of Parasitology, Faculty of Medicine, Universiti Malaya, Kuala Lumpur 50603, Malaysia.
Vector Borne Disease Sector, Ministry of Health, Putrajaya 62000, Malaysia.
Trop Med Infect Dis. 2023 Jul 29;8(8):389. doi: 10.3390/tropicalmed8080389.
The initial and vital stage in the diagnosis of malaria involves extracting DNA. The efficiency of malaria testing is restricted by the multiple steps involved in commercial DNA extraction kits. We attempted to improve an existing loop-mediated isothermal amplification (LAMP) for the detection of by using a simple DNA extraction approach, making it a feasible option for mass screening. We utilized a simple nucleic acid extraction method directly from whole blood for the detection of , taking only 5 min to complete. The extracted DNA was evaluated by two fluorescent-based LAMP and one colorimetric-based LAMP assay. The detection limit for both SYTO-LAMP and SYBR green-LAMP was 0.00001% and 0.0001% parasitemia, respectively. Meanwhile, neutral red-LAMP had a detection limit of 0.01% parasitemia. Combining this simple and inexpensive DNA extraction method, SYTO-LAMP could serve as an alternative molecular diagnosis for the detection of and other human spp.
疟疾诊断的初始关键阶段涉及提取DNA。商业DNA提取试剂盒所包含的多个步骤限制了疟疾检测的效率。我们尝试通过使用一种简单的DNA提取方法来改进现有的环介导等温扩增技术（LAMP）以用于疟原虫检测，使其成为大规模筛查的可行选择。我们采用了一种直接从全血中提取核酸的简单方法来检测疟原虫，整个过程仅需5分钟即可完成。提取的DNA通过两种基于荧光的LAMP检测方法和一种基于比色法的LAMP检测方法进行评估。SYTO-LAMP和SYBR green-LAMP的检测限分别为0.00001%和0.0001%的疟原虫血症。同时，中性红-LAMP的检测限为0.01%的疟原虫血症。结合这种简单且廉价的DNA提取方法，SYTO-LAMP可作为检测疟原虫及其他人体疟原虫种类的替代分子诊断方法。
