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Evaluation of viability PCR performance for assessing norovirus infectivity in fresh-cut vegetables and irrigation water.

Abstract：

:Norovirus (NoV) detection in food and water is mainly carried out by quantitative RT-PCR (RT-qPCR). The inability to differentiate between infectious and inactivated viruses and the resulting overestimation of viral targets is considered a major disadvantage of RT-qPCR. Initially, conventional photoactivatable dyes (i.e. propidium monoazide, PMA and ethidium monoazide, EMA) and newly developed ones (i.e. PMAxx and PEMAX) were evaluated for the discrimination between infectious and thermally inactivated NoV genogroup I (GI) and II (GII) suspensions. Results showed that PMAxx was the best photoactivatable dye to assess NoV infectivity. This procedure was further optimized in artificially inoculated lettuce. Pretreatment with 50μM PMAxx and 0.5% Triton X-100 (Triton) for 10min reduced the signal of thermally inactivated NoV by ca. 1.8 logs for both genogroups in lettuce concentrates. Additionally, this pretreatment reduced the signal of thermally inactivated NoV GI between 1.4 and 1.9 logs in spinach and romaine and lamb's lettuces and by >2 logs for NoV GII in romaine and lamb's lettuce samples. Moreover this pretreatment was satisfactorily applied to naturally-contaminated water samples with NoV GI and GII. Based on the obtained results this pretreatment has the potential to be integrated in routine diagnoses to improve the interpretation of positive NoV results obtained by RT-qPCR.

journal_name

Int J Food Microbiol

journal_title

International journal of food microbiology

authors

Randazzo W,López-Gálvez F,Allende A,Aznar R,Sánchez G

doi

10.1016/j.ijfoodmicro.2016.04.010

subject

Has Abstract

pub_date

2016-07-16 00:00:00

pages

1-6

eissn

0168-1605

issn

1879-3460

pii

S0168-1605(16)30169-6

journal_volume

229

pub_type

杂志文章

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