PAKISTAN
VETERINARY
JOURNAL
     
 
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Development of a Real-Time Quantitative PCR Assay for Detection of Porcine Circovirus Type 2e
 
Xiaohui Wen1,2,3, Dianhong Lv1,2,3, Qi Zhai1,2,3, Xiurong Zhou1,2,3, Mushtaq Ahmad Gondal4, Mudassar Nazar5, Muhammad Usman Saleem6, Chunling Jia1,2,3*, Shao-Lun Zhai1,2,3*
 

1Institute of Animal Health, Guangdong Academy of Agricultural Sciences, Guangzhou, 510640, China; 2Scientific Observation and Experiment Station of Veterinary Drugs and Diagnostic Techniques of Guangdong Province, Ministry of Agriculture of Rural Affairs, Guangzhou, 510640, China; 3Key Laboratory of Animal Disease Prevention of Guangdong Province, Guangzhou, 510640, China; 4University Diagnostic Laboratory, Cholistan University of Veterinary and Animal Sciences Bahawalpur 63100, Bahawalpur, Pakistan; 5University of Agriculture Faisalabad, Sub-Campus Burewala-61010, Pakistan; 6Department of Biosciences, Faculty of Veterinary Sciences, Bahauddin Zakariya University Multan, Pakistan
*Corresponding author: Chunling Jia: jcl898@163.com; Shao-Lun Zhai: zhaishaolun@163.com

Abstract   

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/µL, 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.

To Cite This Article: Wen X, Lv D, Zhai Q, Zhou X, Gondal MA, Nazar M, Saleem MU, Jia C, Zhai SL 2022. Development of a real-time quantitative pcr assay for detection of porcine circovirus type 2e. Pak Vet J, 42(3): 429-432. http://dx.doi.org/10.29261/pakvetj/2020.043

 
   
 

ISSN 0253-8318 (Print)
ISSN 2074-7764 (Online)



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