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Development of a Real-Time Quantitative PCR Assay for Detection of Porcine Circovirus Type 2e

文献类型： 外文期刊

作者： Wen, Xiaohui 1 ; Lv, Dianhong 1 ; Zhai, Qi 1 ; Zhou, Xiurong 1 ; Gondal, Mushtaq Ahmad 4 ; Nazar, Mudassar 5 ; Saleem, Muhammad Usman 6 ; Jia, Chunling 1 ; Zhai, Shao-Lun 1 ;

作者机构： 1.Guangdong Acad Agr Sci, Inst Anim Hlth, Guangzhou 510640, Peoples R China

2.Minist Agr Rural Affairs, Sci Observat & Expt Stn Vet Drugs & Diagnost Tech, Guangzhou 510640, Peoples R China

3.Key Lab Anim Dis Prevent Guangdong Prov, Guangzhou 510640, Peoples R China

4.Cholistan Univ Vet & Anim Sci Bahawalpur, Univ Diagnost Lab, Bahawalpur 63100, Pakistan

5.Univ Agr Faisalabad, Sub Campus, Burewala 61010, Pakistan

6.Bahauddin Zakariya Univ Multan, Fac Vet Sci, Dept Biosci, Multan, Pakistan

关键词： Porcine circovirus type 2e; Quantitative PCR; TaqMan probe; Diagnosis; Epidemiological investigation

期刊名称：PAKISTAN VETERINARY JOURNAL （ 影响因子：1.803； 五年影响因子：1.233 ）

ISSN： 0253-8318

年卷期： 2022 年 42 卷 3 期

页码：

收录情况： SCI

摘要： PCV2e shows high sequence similarity to other PCV2 genotype, causing greater difficulties in the molecular biological identification of pathogens. In this study, we developed a TaqMan-based RT-qPCR for efficient detection of PCV2e in clinical samples. We designed specific primers and a probe for the PCV2e ORF2 gene and then identified target genes using the optimized PCR reaction system and reaction conditions. The results indicated that the cycle threshold (Ct) and the standard DNA template had a linear relationship between 103-108 copies/mu L, and the determination coefficient was 0.998. The assay has a detection limit of 100 copies per reaction and a standard deviation of cycle thresholds below 1.00. The PCR method we established can specifically identify PCV2e, did not cross-react with other genotypes of PCV, as well as PRV and PPV. The RT-qPCR assay can be used for the special diagnosis and epidemiological investigation of PCV2e.

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