title: Evaluation of poliovirus vaccines for pestivirus contamination: non-specific amplification of poliovirus sequences by pan-pestivirus primers.
abstract: Two lots of polyvalent live vaccines for human use against poliovirus were tested by reverse transcriptase (RT) and nested PCR for the presence of contaminating pestivirus RNA. By RT-PCR, samples from both lots showed a band of approximately 450 bp instead of 300 bp for the reference pestivirus strain used as positive control. After nested PCR, the template DNA (450 bp product) was not amplified, suggesting non-specificity of the previous amplification. Sequencing analysis confirmed the non-specificity of the 450 bp bands and revealed, respectively, 80 and 77% homology with a region in the VP1 gene of poliovirus type 1 in samples 1 and 2. This suggests that more caution should be taken in interpreting the results obtained by PCR, and that they should be confirmed by nested PCR or sequencing.
authors.affiliation: Department of Pharmacology, University of Milan, Via Vanvitelli, 32, 20129, Milan, Italy.
authors.forename: Carlo
authors.initials: C
authors.lastname: Zanotto
authors.forename: Massimo
authors.initials: M
authors.lastname: Giangaspero
authors.forename: Mathias
authors.initials: M
authors.lastname: Büttner
authors.forename: Angelika
authors.initials: A
authors.lastname: Braun
authors.forename: Carlo De Giuli
authors.initials: Cde G
authors.lastname: Morghen
authors.forename: Veronica
authors.initials: V
authors.lastname: Elli
authors.forename: Alfonso
authors.initials: A
authors.lastname: Panuccio
authors.forename: Antonia
authors.initials: A
authors.lastname: Radaelli
mesh_terms: D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D017931:DNA Primers; D004340:Drug Contamination; D004341:Drug Evaluation; D000882:Haplorhini; D006801:Humans; D008969:Molecular Sequence Data; D010576:Pestivirus / Q000235:genetics / Q000302:isolation & purification*; D023321:Poliovirus Vaccines; D016133:Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012689:Sequence Homology, Nucleic Acid
chemical_list: D017931:DNA Primers; D023321:Poliovirus Vaccines