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Bioprocessing and integration of a high flux screening systematic platform based on isothermal amplification for the detection on 8 common pathogens
Bioprocess and Biosystems Engineering ( IF 3.8 ) Pub Date : 2020-08-29 , DOI: 10.1007/s00449-020-02423-4
Huamin Zhong 1 , Hongwei Deng 2 , Ming Li 2 , Huahong Zhong 2
Affiliation  

During a large variety of common pathogens, E. coli, P. aeruginosa, MRSA, MRCNS, V. parahaemolyticus, L. monocytogenes and Salmonella are the leading pathogens responsible for large number of human infections and diseases. In this study, a high flux screening based on nucleic acid isothermal amplification technique has been developed. For the 8 common pathogens, species-specific targets had been selected and analyzed for their unique specificity. After optimization, separate LAMP reaction assays had been bioprocessed and integrated into one systematic detection platform, including 8 strips (PCR tubes) and 96-well plates. Eight standard strains verified for the accuracy. Application of the established high flux screening platform was used for detection for 48 samples in 4 different 96-well plates, with 2 groups of 2 operators using double-blind procedure. The accuracy of 100% was obtained, with the total time consumption as 66–75 min (for 12 samples detection on 8 different pathogens). As concluded, through the bioprocess of the systematic platform based on LAMP technique, it’s been demonstrated to be capable of simultaneous detection of 8 pathogens, with high sensitivity, specificity, rapidity and convenience.



中文翻译:

基于等温扩增的8种常见病原菌高通量筛选系统平台的生物处理与集成

在多种常见病原体期间,大肠杆菌铜绿假单胞菌、MRSA、MRCNS、副溶血性弧菌单核细胞增生李斯特菌沙门氏菌是导致大量人类感染和疾病的主要病原体。在这项研究中,开发了一种基于核酸等温扩增技术的高通量筛选。对于 8 种常见病原体,已选择物种特异性靶标并分析其独特的特异性。优化后,单独的 LAMP 反应测定已被生物处理并集成到一个系统检测平台中,包括 8 个条带(PCR 管)和 96 孔板。八种标准菌株验证了准确性。应用建立的高通量筛选平台对4个不同的96孔板中的48个样品进行检测,两组2人采用双盲程序。获得了 100% 的准确率,总时间消耗为 66-75 分钟(用于 8 种不同病原体的 12 个样品检测)。综上所述,通过基于LAMP技术的系统平台的生物过程,已证明能够同时检测8种病原体,具有高灵敏度、特异性、快速性和便捷性。

更新日期:2020-08-29
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