๐ Development of a SYBR green-based real-time RT-PCR assay for rapid detection of the emerging swine acute diarrhea syndrome coronavirus
Swine acute diarrhea syndrome coronavirus (SADS-CoV) is a novel coronavirus which was associated with severe diarrhea disease in pigs. SADS-CoV was first detected and identified as the causative agent of a devastating swine disease outbreak in southern China in 2017. Routine monitoring and early detection of the source of infection is therefore integral to the prevention and control of SADS-CoV infection. In this study, a SYBR green-based real-time reverse transcription quantitative polymerase chain reaction (RT-qPCR) technique was established for rapid detection and monitoring of this emerging virus. Specific primers were designed based on the conserved region within the M gene of the viral genome. The lowest detection limit of the RT-qPCR assay was 10 copies/ฮผL. This assay was specific and had no cross-reaction with other 11 swine viruses. The positive rate of 84 clinical samples for the SYBR green-based RT-qPCR and the conventional RT-PCR was 73.81% (62/84) and 53.57% (45/84), respectively. These results demonstrated that the SYBR green-based RT-qPCR technique was an effectively diagnostic method with higher sensitivity than probe-based RT-qPCR and gel-based RT-PCR for detection and epidemiological investigations of SADS-CoV.
keywords
๐ syndrome coronavirus (1074)
๐ causative agent (117)
๐ reverse transcription (205)
๐ polymerase chain (300)
๐ novel coronavirus (684)
๐ viral genome (96)
๐ real-time reverse (87)
๐ chain reaction (303)
author
๐ค Ma, Lei
๐ค Zeng, Fanwen
๐ค Cong, Feng
๐ค Huang, Bihong
๐ค Huang, Ren
๐ค Ma, Jingyun
๐ค Guo, Pengju
year
โฐ 2019
issn
๐ 18790984 01660934
volume
265
number
page
66-70
citedbycount
1
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