Real-time PCR protocol for the detection of porcine parvovirus in field samples.
This report describes a real-time polymerase chain reaction assay with SYBR Green for detection of a broad range of porcine parvoviruses (PPV) and accurate virus quantification in porcine tissues. The assay targets the VP2 gene of PPV and the porcine genomic c-myc gene for normalization. The detection limit of the SYBR Green reaction was shown to be equivalent to 6 x 10(0) to 6 x 10(1) PPV copies/reaction and the overall detection limit equivalent to 0.1 TCID(50)/100 microl. The assay was linear over a 10(7) dilution range of template concentrations. Other porcine pathogens involved in reproductive disorders such as porcine circovirus 2 (PCV-2), porcine reproductive and respiratory virus (PRRSV), Aujeszky's disease virus (PRV) and other parvoviruses such as feline parvovirus (FPV), canine parvovirus (CPV), minute virus of canines (MVC) and a human parvovirus (B19) were not detected by this assay.
Wilhelm
Sonja
S
Institute for Animal Hygiene and Veterinary Public Health, An den Tierkliniken 1, 04103 Leipzig, Germany.
Zimmermann
Pia
P
Selbitz
Hans Joachim
HJ
Truyen
Uwe
U
D000818:Animals; D000835:Animals, Wild; D052160:Benzothiazoles; D036022:Capsid Proteins / Q000235:genetics; D002460:Cell Line; D017931:DNA Primers; D003959:Diamines; D005333:Fetus; D005814:Genes, Viral; D016259:Genes, myc / Q000235:genetics; D009930:Organic Chemicals; D010322:Parvoviridae Infections / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D028323:Parvovirus, Porcine / Q000235:genetics / Q000302:isolation & purification*; D011804:Quinolines; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013552:Swine; D013553:Swine Diseases / Q000175:diagnosis / Q000821:virology; D010948:Viral Plaque Assay / Q000379:methods; D014781:Viscera / Q000821:virology
D052160:Benzothiazoles; D036022:Capsid Proteins; D017931:DNA Primers; D003959:Diamines; D009930:Organic Chemicals; D011804:Quinolines; C098022:SYBR Green I