Mao Rui, Ge Ge, Wang Zhuo, Hao Rongzhang, Zhang Guohao, Yang Zhenzhou, Lin Bincheng, Ma Yajun, Liu Hongtao, Du Yuguang
Key Laboratory of Biopharmaceutical Production & Formulation Engineering, PLA and State Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100190, PR China; University of Chinese Academy of Sciences, Beijing 100049, PR China.
Key Laboratory of Biopharmaceutical Production & Formulation Engineering, PLA and State Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100190, PR China.
Acta Trop. 2018 Feb;178:86-92. doi: 10.1016/j.actatropica.2017.10.025. Epub 2017 Nov 2.
Malaria infection poses a great threaten to public health even nowadays. The conventional diagnosis tools of malaria parasites and vectors require systematic training for the observers accompanied by the low throughput. In this study, a new detection system, i.e., multiplex microfluidic loop-mediated isothermal amplification (mμLAMP) array, was developed to provide a convenient, rapid and economical detection system for malaria diagnosis. A microfluidic-based detection chip was designed and developed, targeting the conserved gene of four Anopheles and two Plasmodium species responsible for most of the malaria cases occurred in China. The DNA preparation of Anopheles and Plasmodium samples was realized by using a newly-developed DNA extraction method. For this mμLAMP array system, the detection limit was determined to be 1pg of targeting DNA with high sensitivity (>95%) and specificity (100%). Further, the accuracy of such mμLAMP analysis was evaluated by the analysis of 48 Anopheles mosquito samples, of which 30 were termed to be target Anopheles, displaying high consistency with that by morphological analysis. In conclusion, the mμLAMP detection system was proved to be a visible, sensitive, specific and high-throughput diagnostic tool. Considering the portable manipulation of such detection system, our studies shed light on its potential application of malaria surveillance on the spot.
即使在当今，疟疾感染对公共卫生仍构成巨大威胁。传统的疟原虫和病媒诊断工具需要对观察者进行系统培训，且通量较低。在本研究中，开发了一种新的检测系统，即多重微流控环介导等温扩增（mμLAMP）阵列，以提供一种方便、快速且经济的疟疾诊断检测系统。设计并开发了一种基于微流控的检测芯片，其靶向在中国导致大多数疟疾病例的四种按蚊和两种疟原虫物种的保守基因。通过使用新开发的DNA提取方法实现了按蚊和疟原虫样本的DNA制备。对于该mμLAMP阵列系统，检测限确定为1pg靶向DNA，具有高灵敏度（>95%）和特异性（100%）。此外，通过对48个按蚊样本的分析评估了这种mμLAMP分析的准确性，其中30个被确定为目标按蚊，与形态学分析显示出高度一致性。总之，mμLAMP检测系统被证明是一种可视化、灵敏、特异且高通量的诊断工具。考虑到这种检测系统便于携带操作，我们的研究为其在疟疾现场监测中的潜在应用提供了思路。
