环介导等温扩增
生物过程
工业与生产工程
生物
微生物学
核酸
计算生物学
DNA
电气工程
遗传学
工程类
古生物学
生物化学
作者
Huamin Zhong,Hongwei Deng,Ming Li,Huahong Zhong
标识
DOI:10.1007/s00449-020-02423-4
摘要
Abstract 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.
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