Evaluation of real-time RT-PCR assay for the routine intra vitam diagnosis of classical swine fever.
The aim of the study was to evaluate the diagnostic quality of the real-time RT-PCR assay described by Hoffmann et al. [Hoffmann, B., Beer, M., Schelp, C., Schirrmeier, H., Depner, K., 2005. Validation of a real-time RT-PCR assay for sensitive and specific detection of classical swine fever. J. Virol. Methods 130, 36-44] for the routine intra vitam diagnosis of classical swine fever (CSF). We compared the assay with conventional diagnostic methods by using defined diagnostic material from an animal experiment with pigs showing different clinical forms of CSF. Compared to virus isolation and antigen ELISA an enhanced sensitivity of the real-time RT-PCR could be shown. We were able to detect all infected pigs regardless of the clinical course of CSF. CSF infection was detected already during the incubation period, during the entire clinical phase as well as at the beginning of convalescence when the first antibodies were detected and no virus could be isolated any more. In most cases, positive PCR results were obtained 2 days earlier than with virus isolation and 2-4 days earlier than with the antigen ELISA.
Depner
Klaus
K
Friedrich-Loeffler-Institut, Institute of Diagnostic Virology, Boddenblick 5a, 17493 Greifswald-Insel Riems, Germany. Klaus.depner@fli.bund.de
Hoffmann
Bernd
B
Beer
Martin
M
D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D000956:Antigens, Viral / Q000097:blood; D006691:Classical Swine Fever / Q000175:diagnosis* / Q000821:virology; D006692:Classical Swine Fever Virus / Q000235:genetics / Q000302:isolation & purification*; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary; D009500:Neutralization Tests / Q000662:veterinary; D012367:RNA, Viral / Q000737:chemistry / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary*; D012680:Sensitivity and Specificity; D013552:Swine; D014766:Viremia / Q000097:blood / Q000662:veterinary* / Q000821:virology
D000914:Antibodies, Viral; D000956:Antigens, Viral; D012367:RNA, Viral