Detection ofpestivirus contamination in cellcultures bynested-PCR

S. A. Ghorashi, M. Daliri Joupari, D. Morshedi, T. Hajian, M. Lotfi

Research output: Contribution to journalArticle

Abstract

In this study a nested-PCR assay was optimized for detection of two BVDV biotype of NADL strain. Apart 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 10 4 TCID50 and 10 2 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.

Original languageEnglish
Pages (from-to)11-16
Number of pages6
JournalJournal of Veterinary Research
Volume63
Issue number1
Publication statusPublished - 01 May 2008

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