Temperature inactivation of Feline calicivirus vaccine strain FCV F-9 in comparison with human noroviruses using an RNA exposure assay and reverse transcribed quantitative real-time polymerase chain reaction-A novel method for predicting virus infectivity

被引：96

作者：

Topping, J. R. ^{[1
]}

Schnerr, H. ^{[1
]}

Haines, J. ^{[1
]}

Scott, M. ^{[1
]}

Carter, M. J. ^{[2
]}

Willcock, M. M. ^{[2
]}

Bellamy, K. ^{[3
]}

Brown, D. W. ^{[4
]}

Gray, J. J. ^{[4
]}

Gallimore, C. I. ^{[4
]}

Knight, A. I. ^{[1
]}

机构：

[1] Leatherhead Food Int, Leatherhead KT22 7RY, Surrey, England

[2] Univ Surrey, Fac Hlth & Med Sci, Surrey GU2 7XH, England

[3] Unilever Cent Resources Ltd, Sharnbrook MK44 1LQ, Beds, England

[4] Hlth Protect Agcy, Ctr Infect, London NW9 5EQ, England

来源：

JOURNAL OF VIROLOGICAL METHODS | 2009年 / 156卷 / 1-2期

关键词：

Norovirus; NoVs; Calicivirus; RNase; RT-QPCR; Inactivation; Virus infectivity; ROUND STRUCTURED VIRUSES; PCR; ENVIRONMENT; STABILITY; SURROGATE; REVEALS;

D O I：

10.1016/j.jviromet.2008.10.024

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

A one-step reverse transcription quantitative real-time polymerase chain reaction (RT-QPCR) method in combination with RNase treatment and low copy number samples was developed in order to examine the effect of temperature on the ability of virus capsids to protect their RNA content. The method was applied to a non-cultivable virus (GII.4 norovirus) and Feline calicivirus vaccine strain F-9 (FCV) which is often used as a norovirus surrogate. Results demonstrated that FCV RNA is exposed maximally after 2 min at 63.3 degrees C and this correlated with a greater than 4.5 log reduction in infectivity as assessed by plaque assay. In contrast human GII.4 norovirus RNA present in diluted clinical specimens was not exposed maximally until 76.6 degrees C, at least 13.3 degrees C greater than that for FCV. These data suggest that norovirus possesses greater thermostability than this commonly used surrogate. Further, these studies indicate that current food processing regimes for pasteurisation are insufficient to achieve inactivation of GII.4 NoVs. The method provides a novel molecular method for predicting virus infectivity. (c) 2008 Elsevier B.V. All rights reserved.

引用

页码：89 / 95

页数：7