|Callahan, Johnny - TETRACORE, INC., MD|
|Nelson, William - TETRACORE, INC., MD|
Submitted to: Journal of Clinical Microbiology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: November 3, 2002
Publication Date: January 5, 2003
Interpretive Summary: Classical Swine Fever (CSF), also known as Hog Cholera, is a highly infectious, sometimes fatal, disease of domestic and wild pigs, caused by Classical Swine Fever Virus (CSFV). CSF is distributed worldwide, including South and Central America, southern Mexico and the Caribbean, thus posing a serious threat to the pork industry of countries like US that are free of the disease. CSF can spread rapidly in areas with high pork density and free of the disease, thus rapid and precise detection of CSFV, preferably on site, is critical for disease containment. To address this need we have developed a real time diagnostic test based on PCR TaqMan technology for CSFV detection. The test is specific for CSFV, highly sensitive, and detects isolates from different geographical areas. Importantly the assay detects the virus before the appearance of clinical signs when tested in experimentally infected animals. The assay is performed in a single tube, containing all diied reagents, and performed in less than 2 hours, on a portable detection instrument in real time. The simplicity and portability of the assay allow for its use as a pen-side assay for rapid on-site detection of CSFV
Technical Abstract: A fluorogenic probe hydrolysis (TaqMan)-reverse transcriptase PCR assay for CSF virus (CSFV) was developed and evaluated in experimentally infected swine. The assay detected CSFV representing different phylogenetic groupings but did not amplify viral RNA from related Pestiviruses. The assay met or exceeded the sensitivity (1-100 tissue culture infective doses per ml) of viral cultures on samples from experimentally infected animals. Viral RNA was detected in nasal and tonsil scraping samples 2 to 4 days prior to onset of clinical disease. The assay can be performed in two hours or less, thus providing a rapid method for the diagnosis of CSF.