Identification of inhibition of protein disulphide isomerase expression related to classical swine fever virus infection by using real-time PCR analysis

Abstract

Although protein disulphide isomerase (PDI) is a well-known multifunctional protein, the precise mechanism of its response to viral infections has not been fully elucidated, especially in veterinary research. Therefore, in this study we developed a real-time polymerase chain reaction (PCR) method for the simultaneous detection of PDI expression in porcine cell lines and tissues. Several primers were designed to amplify a specific nucleotide sequence of PDI and to develop a PCR-based biotechnology platform in macrophages. The sensitivity of the assay for detecting PDI was determined to be in the range of 108–101 genomic units. Our results demonstrated that this method could be used to accurately quantify PDI expression and to examine its effects on infection responses to classical swine fever virus (CSFV) in porcine tissue. PDI expression was found to be inhibited in the heart, liver, spleen, lung, kidney and mesenteric lymph node tissue from a CSFV-positive pig. These data contribute to an improved understanding of the mechanism of CSFV pathogenesis.