Simultaneous detection and differentiation of porcine circovirus 3 and 4 using a SYBR Green I-based duplex quantitative PCR assay

被引：16

作者：

Hou, Cheng-Yao ^{[1
]}

Xu, Tong ^{[1
]}

Zhang, Liu-Hui ^{[1
]}

Cui, Jian-Tao ^{[1
]}

Zhang, Yuan-Hang ^{[1
]}

Li, Xin-Sheng ^{[1
]}

Zheng, Lan-Lan ^{[1
]}

Chen, Hong-Ying ^{[1
]}

机构：

[1] Henan Agr Univ, Zhengzhou Key Lab Pig Dis Prevent & Control, Coll Vet Med, Zhengdong New Dist Longzi Lake 15, Zhengzhou 450046, Henan, Peoples R China

来源：

JOURNAL OF VIROLOGICAL METHODS | 2021年 / 293卷

关键词：

Porcine circovirus 3; Porcine circovirus 4; Duplex quantitative PCR; GENETIC-CHARACTERIZATION; TYPE-3; PATHOGENESIS; DISEASE; ORIGIN; PIGS;

D O I：

10.1016/j.jviromet.2021.114152

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Porcine circovirus 4 (PCV4) was a novel circovirus identified from diseased pigs in 2019 in Hunan Province, China, and PCV3 and PCV4 co-infection has been reported. In order to detect and differentiate PCV3 and PCV4 simultaneously, the SYBR Green.-based duplex quantitative PCR (qPCR) assay was established in the present study. The two viruses could be easily distinguished by different Tm values: 86.5 degrees C for PCV3 and 79 degrees C for PCV4, while other porcine pathogens did not shown specific melting peaks. The detection limits of this duplex qPCR assay were 51.7 copies/mu L for PCV3 and 67.7 copies/mu L for PCV4, and both of the intra-assay and inter-assay of the CV analysis of this assay were less than 2.0 %. Sixty-four clinical samples from 22 different swine farms were screened by the duplex qPCR assay. The results showed that the positive detection rate of PCV3 was 37.5 % (24/ 64) and PCV4 was 34.38 % (22/64), and PCV3 and PCV4 co-infection rate was 17.19 % (11/64). The detection rate of the duplex qPCR assay was higher than that of the conventional PCR assay. The duplex qPCR was of high sensitivity and specificity, being able to provide technical support for clinical detection, differential diagnosis and control of PCV3 and PCV4.

引用

页数：5

共 27 条

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