Development of SYBR Green I-based quantitative PCR assay for identification of porcine circovirus 1, 2 and 3
文献类型： 外文期刊
作者： Liu, Chang 1 ; Tian, Yao 1 ; Zheng, Li-mei 2 ; Ma, Ying-Ru 2 ; Shi, Jian-Li 1 ; Li, Chen 1 ; Xu, Shao-Jian 1 ; Wu, Xiao-Yan 1 ; Han, Hong 1 ; Li, Jun 1 ; Han, Xian-Jie 2 ;
作者机构： 1.Shandong Acad Agr Sci, Shandong Prov Key Lab Anim Dis Control & Breediin, Inst Anim Sci & Vet Med, 202 Ind North Rd, Jinan 250100, Peoples R China
2.Qingdao Agr Univ, Coll Vet Med, Qingdao, Peoples R China
关键词： porcine circovirus; specificity; sensitivity; SYBR Green I based quantitative PCR; single identification method
期刊名称：VETERINARSKI ARHIV （ 影响因子：0.5； 五年影响因子：0.6 ）
ISSN： 0372-5480
年卷期： 2023 年 93 卷 4 期
页码：
收录情况： SCI
摘要： Porcine Circovirus (PCV) includes Porcine Circovirus 1(PCV1), Porcine Circovirus 2 (PCV2) and Porcine Circovirus 3 (PCV3). In recent years, co-infection exists between PCV1, PCV2 and PCV3 serotypes. Therefore, it is particularly necessary to establish a fast, specific and sensitive SYBR Green I real-time quantitative PCR detection method for PCV1, PCV2 and PCV3. In this experiment, specific primers were selected and the reaction conditions were optimized. A real-time quantitative PCR identification method was established. The results showed the detection limits of this assay were 40.3 copies/mu l for PCV1, 25.2 copies/mu l for PCV2 and22.4 copies/mu l for PCV3. There was no cross-reactivity with swine fever virus (CSFV), porcine reproductive and respiratory syndrome virus (PRRSV), porcine pseudorabies virus (PRV) and porcine parvovirus (PPV). The intra-assay and inter-assay coefficients of variation were less than 1%. The test results of 100 PCV suspected positive samples revealed that the PCV1, PCV2 and PCV3 singular infection rate was 10% (10/100), 64% (64/100) and 52% (52/100), respectively. The PCV1 and PCV2 co-infection rate was 8% (8/100), the PCV1 and PCV3 co-infection rate was 7% (7/100), the PCV2 and PCV3 co-infection rate was 26% (26/100), and the PCV1, PCV2 and PCV3 co-infection rate was 7% (7/100). This method has good specificity, sensitivity and stability. It provides a promising tool for rapid differential detection of PCV1, PCV2 and PCV3.
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