detection of pestivirus contamination in cell cultures by nested-pcr

in this study a nested-pcr assay was optimized for detection of two bvdv biotype of nadl strain. a part of 5' non-coding region of virus, 249 bp in size, was amplified in rt-pcr. pcr product was cloned in a ptz57r/t vector and sequencing results confirmed the specificity of the test. internal primers were designed and a 155 bp dna fragment was amplified in nested-pcr. the sensitivity of rt-pcr and nested-pcr for detection of virus in cell culture were found to be 104 tcid50 and 102 tcid50, respectively. seven cell cultures were tested for bvdv contamination using elisa, rt-pcr and nested-pcr. results indicate that sensitivity of molecular tests for detection of virus in cell culture samples is higher than elisa.