Evaluation of poliovirus vaccines for pestivirus contamination:: non-specific amplification of poliovirus sequences by pan-pestivirus primers

被引:4
作者
Zanotto, C
Giangaspero, M
Büttner, M
Braun, A
Morghen, CD
Elli, V
Panuccio, A
Radaelli, A
机构
[1] Univ Milan, Dept Pharmacol, I-20129 Milan, Italy
[2] Univ Milan, Inst Special Pathol, Fac Vet Med, Milan, Italy
[3] Univ Milan, Vet Med Clin, Milan, Italy
[4] Fed Res Ctr Virus Dis Anim, Inst Immunol, D-7400 Tubingen, Germany
[5] Univ Milan, Dept Pharmacol Sci, Milan, Italy
[6] ASL City Milan, Publ Hlth Lab, Milan, Italy
[7] CNR, Cellular & Mol Pharmacol Ctr, I-20133 Milan, Italy
关键词
vaccines; pestivirus contamination; RT-PCR; nested PCR;
D O I
10.1016/S0166-0934(02)00009-5
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
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. (C) 2002 Published by Elsevier Science B.V.
引用
收藏
页码:167 / 172
页数:6
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