The Colorimetric Isothermal Multiple-Self-Matching-Initiated Amplification Using Cresol Red for Rapid and Sensitive Detection of Porcine Circovirus 3
文献类型： 外文期刊
作者： Gou, Hongchao 1 ; Bian, Zhibiao 1 ; Cai, Rujian 1 ; Jiang, Zhiyong 1 ; Song, Shuai 1 ; Li, Yan 1 ; Chu, Pinpin 1 ; Yang, 1 ;
作者机构： 1.Guangdong Acad Agr Sci, Inst Anim Hlth, Guangzhou, Peoples R China
2.Guangdong Prov Key Lab Livestock Dis Prevent, Guangzhou, Peoples R China
3.Guangdong Open Lab Vet Publ Hlth, Guangzhou, Peoples R China
4.Sci Observat & Expt Stn Vet Drugs & Diagnost Tech, Guangzhou, Peoples R China
5.Zhongkai Univ Agr & Engn, Coll Anim Sci & Technol, Guangzhou, Peoples R China
关键词： PCV3; isothermal multiple-self-matching-initiated amplification (IMSA); detection; colorimetric assay; cresol red
期刊名称：FRONTIERS IN VETERINARY SCIENCE （ 影响因子：3.412； 五年影响因子：3.588 ）
ISSN：
年卷期： 2020 年 7 卷
页码：
收录情况： SCI
摘要： In 2016, a novel porcine circovirus (PCV), PCV3, was identified in USA. Subsequently, it was proved to be also epidemic in China, Poland, and Korea. To analyze and control the epidemic situation of PCV3, it is necessary to establish accurate and high-throughput detection methods. In this study, the colorimetric isothermal multiple-self-matching-initiated amplification (IMSA) using cresol red was developed to detect PCV3 for the first time. The reaction can be easily performed by incubating the tube at 63 degrees C for 60 min. By the addition of pH-sensitive indicator dye cresol red, the initial color of the reaction mixture is red. When PCV3 capsid gene DNA was positive in the sample, the color of the reaction mixture changed from red to yellow after the isothermal incubation at 63 degrees C, while the negative control maintained the red color. The colorimetric IMSA displayed good specificity in detecting PCV3, PCV2, and PCV1 and 4 porcine DNA pathogens. Moreover, it has a low and repeatable detection limit of 10 copies, which is consistent with TaqMan-based qPCR, but 10 times more sensitive than PCR. In diagnosing 128 clinical specimens, it not only showed 100% agreement with qPCR but also detected 15 positive results more than PCR. The colorimetric IMSA we offered might be a good choice for PCV3 epidemiological investigation and point-of-care testing.
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