Journal article
Rapid and sensitive detection of type II porcine reproductive and respiratory syndrome virus by reverse transcription loop-mediated isothermal amplification combined with a vertical flow visualization strip
H Gou, J Deng, J Pei, J Wang, W Liu, M Zhao, J Chen
Journal of Virological Methods | ELSEVIER SCIENCE BV | Published : 2014
Abstract
Reverse transcription-loop-mediated isothermal amplification (RT-LAMP) was combined with a vertical flow (VF) nucleic acid detection strip to develop a universal assay for the detection of type II porcine reproductive and respiratory syndrome virus (PRRSV). The loop primers were labeled separately with biotin and fluorescein isothiocyanate (FITC) in this assay. Using optimized parameters, the whole reaction could be completed in <50min in a completely enclosed environment. The detection limit of this assay was found to be 1pg RNA, 30 tissue culture infective dose 50 (TCID50) virus, or 230 copies of recombinant plasmid DNA, which is relatively higher than that of RT-LAMP analyzed by agarose g..
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Awarded by National Natural Science Foundation of China
Funding Acknowledgements
This research was supported by grants from the Special Fund for Agro-Scientific Research in the Public Interest (No. 201203056), the National Natural Science Foundation of China (Nos. 31172321 and 31472200), the Special Project for Scientific and Technological Innovation in Higher Education of Guangdong, China (No. 2012CXZD0013), and the (Integration of Production, Teaching and Research) Project of Department of Education of Guangdong Province (No. 20118090400259).