Establishment of a duplex fluorescent quantitative PCR assay for detection of Porcine circovirus type 2 and type 3
Acta Agriculturae Zhejiangensis
; 34(3):457-463, 2022.
Article
in Chinese
| CAB Abstracts | ID: covidwho-20240064
ABSTRACT
To establish a method for simultaneous detection of porcine circovirus type 2 (PCV2) and porcine circovirus type 3 (PCV3), specific primers and TaqMan probes were designed after sequence alignment according to the specific sequences of PCV2 Cap gene and PCV3 Cap gene on GenBank. By optimizing the reaction conditions, a duplex fluorescence quantitative PCR detection method for simultaneous detection of porcine circovirus type 2 and 3 was established, and the specificity, sensitivity, and reproducibility were tested. Specificity test results showed that in addition to the positive test results for PCV2 and PCV3, tests for PRRSV, CSFV, PPV, PRV, PEDV, and TGEV were all negative with no cross-reaction, indicating its good specificity. Sensitivity test results showed that the minimum detection limit for detection of PCV2 and PCV3 can both reach 10 copies.L-1, indicating its high sensitivity. The coefficient of variation within and between groups of this method was less than 2%, indicating its good stability. A total of 181 pork and whole blood samples collected from Zhejiang Province were tested using the detection method established in this article and the standard common fluorescent PCR detection method. The results showed that the positive rate of PCV2 was 50.83% (92/181), the positive rate of PCV3 was 37.57% (68/181), and the co-infection rate of PCV2 and PCV3 was 12.15% (22/181). The above detection results of ordinary fluorescent PCR were 50.28% (91/181), 36.46% (66/181), and the co-infection rate was 11.60% (21/181). The coincidence rates of the two methods for PCV2 and PCV3 can reach 98.91% and 97.06%, and the coincidence rate for PCV2 and PCV3 mixed infection were 95.45%. In summary, the duplex fluorescence quantitative PCR detection method established in this experiment can distinguish PCV2 and PCV3 rapidly, which can be used for pathogen detection and epidemiological investigation.
Diagnosis of Animal Diseases [LL886]; Prion; Viral; Bacterial and Fungal Pathogens of Animals [LL821]; Animal Immunology [LL650]; Genetics and Molecular Biology of Microorganisms [ZZ395]; Molecular Biology and Molecular Genetics [ZZ360]; porcine circoviruses; diagnostic techniques; detection; polymerase chain reaction; diagnosis; nucleotide sequences; gene banks; swine diseases; viral diseases; porcine reproductive and respiratory syndrome; Porcine circovirus 2; Porcine circovirus 3; pigs; Porcine reproductive and respiratory syndrome virus; Classical swine fever virus; Porcine parvovirus; Porcine epidemic diarrhea virus; Transmissible gastroenteritis virus; Suid herpesvirus 1; Circovirus; Arterivirus; Varicellovirus; Circoviridae; ssDNA Viruses; DNA Viruses; viruses; Sus scrofa; Sus; Suidae; Suiformes; Artiodactyla; mammals; vertebrates; Chordata; animals; eukaryotes; Arteriviridae; Nidovirales; positive-sense ssRNA Viruses; ssRNA Viruses; RNA Viruses; Pestivirus; Flaviviridae; Parvovirus; Parvovirinae; Parvoviridae; Alphacoronavirus; Coronavirinae; Coronaviridae; Alphacoronavirus 1; Alphaherpesvirinae; Herpesviridae; Herpesvirales; dsDNA Viruses; Porcine circovirus-2; PCR; DNA sequences; swine; hogs; germplasm banks; pig diseases; viral infections; Swine fever virus; Hog cholera virus; hog cholera; Porcine epidemic diarrhoea virus; Aujeszky virus
Full text:
Available
Collection:
Databases of international organizations
Database:
CAB Abstracts
Type of study:
Diagnostic study
/
Experimental Studies
/
Randomized controlled trials
Language:
Chinese
Journal:
Acta Agriculturae Zhejiangensis
Year:
2022
Document Type:
Article
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