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 asPRV
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