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Title: Application of an Improved Micro-amount of Virion Enrichment Technique (MiVET) for the Detection of Avian Influenza A Virus in Spiked Chicken Meat Samples
Authors: Makino, Ryohei
Yamazaki, Yasuko
Nagao, Konomu
Apego, Francis Victor
Mekata, Hirohisa
Yamazaki, Wataru  kyouindb  KAKEN_id  orcid https://orcid.org/0000-0002-7874-8014 (unconfirmed)
Author's alias: 山崎, 安子
山崎, 渉
Keywords: MiVET
Micro-amount of virion enrichment technique
Influenza A virus
Chicken
Meat
Issue Date: Jun-2020
Publisher: Springer Science and Business Media LLC
Journal title: Food and Environmental Virology
Volume: 12
Start page: 167
End page: 173
Abstract: Highly sensitive detection of pathogens is effective for screening meat during quarantine inspection and export. The “micro-amount of virion enrichment technique” (MiVET) was recently developed, which is a new method combining virus concentration with immunomagnetic beads and simple RNA extraction with sodium dodecyl benzenesulfonate (SDBS) for the specific and sensitive detection of avian influenza viruses (AIVs). AIV subtypes H3N2 and H4N2 were used to spike the surface of chicken breast meat samples. The modified MiVET protocol was tested by comparing it against three different homogenate preparation conditions, as well as in samples with added α-amylase and collagenase to digest inhibitors. The performance of the modified MiVET was evaluated by real-time RT-PCR assay targeting the matrix gene. Compared with conventional RNA extraction, the modified MiVET reproducibly concentrated AIVs in chicken meat samples with 100–1000-fold improvement by 60 s-hand homogenization. The 30 s- and 60 s-stomacher homogenizations resulted 100-fold and 10–100-fold improvement, respectively. The modified MiVET required < 60 min from homogenate preparation to final RNA elution. Further, use of the modified MiVET also decreased the rate of false-negative results. The modified MiVET is effective for the rapid and highly sensitive detection of AIVs in chicken meat samples, and can be applied to quarantine and export inspection at airports and seaports.
Rights: This is a post-peer-review, pre-copyedit version of an article published in Food and Environmental Virology. The final authenticated version is available online at: http://dx.doi.org/10.1007/s12560-020-09425-1 .
The full-text file will be made open to the public on 19 March 2021 in accordance with publisher's 'Terms and Conditions for Self-Archiving'.
This is not the published version. Please cite only the published version. この論文は出版社版でありません。引用の際には出版社版をご確認ご利用ください。
URI: http://hdl.handle.net/2433/250998
DOI(Published Version): 10.1007/s12560-020-09425-1
PubMed ID: 32193766
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