Real-time PCR-based infectivity assay for the titration of turkey hemorrhagic enteritis virus, an adenovirus, in live vaccines

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• 作者：Hassan M. Mahsoub^a ; ^b ; ^c ; ^{hmahsoub@vt.edu} ; Nicholas P. Evans^a ; ^c ; ^{nievans@vt.edu} ; Nathan M. Beach^c ; ^{beachnm@gmail.com} ; Lijuan Yuan^d ; ^lyuan@vt.edu ; Kurt Zimmerman^d ; ^{kzimmerm@vt.edu} ; Frank W. Pierson^a ; ^c ; ^{pierson@vt.edu}
• 关键词：CC ; cell culture ; CCID50 ; cell-culture infectious dose 50 ; CLM ; complete Leibovitz&rsquo ; s L15-McCoy&rsquo ; s 5A media ; CPE ; cytopathic effect ; Ct ; threshold cycle ; EPD ; end-point dilution assay ; HE ; hemorrhagic enteritis disease ; CC HE ; cell-culture hemorrhagic enteritis vaccine ; IVP ; infectious viral particle(s) ; LOQ ; limit of quantification ; %CV ; percentage coefficient of variation ; qPCR ; quantitative (real-time) polymerase chain reaction ; SRLM ; serum-reduced Leibovitz&rsquo ; s L15-McCoy&rsquo
• 刊名：Journal of Virological Methods
• 出版年：2017
• 出版时间：January 2017
• 年：2017
• 卷：239
• 期：Complete
• 页码：42-49
• 全文大小：1172 K
• 卷排序：239

文摘

A new infectivity titration method combining qPCR with cell culture was developed for turkey hemorrhagic enteritis virus. The method was used for titration of 9 lots of hemorrhagic enteritis vaccine. Significant variations in genomic and infectious titers were found. Genomic titer of HE vaccines can serve as an indicator of qPCR infectious titer. The method is a useful alternative for current traditional methods.