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Development and evaluation of a droplet digital PCR assay for the detection of fowl adenovirus serotypes 4 and 10 in attenuated vaccines.

Abstract：

:In recent years, there has been an increase in reported cases of fowl adenovirus serotype 4 (FAdV-4) in chickens in China. The use of live attenuated vaccines contaminated with FAdV-4 has been proved to be one of the important causes of massive outbreaks of hydropericardium syndrome. To detect the contamination with FAdV-4 in attenuated vaccines more promptly and accurately, a droplet digital PCR (ddPCR) assay was developed for the rapid detection of FAdV-4 and FAdV-10. The ability of this assay to detect FAdV-4 contamination in attenuated Newcastle disease virus vaccines was assessed in comparison to a quantitative real-time PCR (qPCR) and a conventional PCR assay. The findings indicated that the ddPCR assay could detect FAdV-4 contamination at 0.1 EID50/1,000 feathers, while the qPCR could detect FAdV-4 contamination at 1 EID50/1,000 feathers with identical genomic targets, which was 1,000-fold more sensitive than conventional PCR detection with a sensitivity of 102 EID50/1,000 feathers. The ddPCR assay also showed high specificity for FAdV-4/10 and no positive signals were detected for other FAdVs. Consequently, the intuitive and rapid results were especially suitable for the detection of FAdV-4 contamination in vaccines. In this study, a ddPCR assay was developed to effectively detect and quantify low-dose FAdV-4 contamination, providing a new method for rapid detection of FAdV-4 contamination in various samples, especially vaccines.

journal_name

J Virol Methods

journal_title

Journal of virological methods

authors

Dong G,Meng F,Zhang Y,Cui Z,Lidan H,Chang S,Zhao P

doi

10.1016/j.jviromet.2018.09.005

subject

Has Abstract

pub_date

2019-03-01 00:00:00

pages

59-65

eissn

0166-0934

issn

1879-0984

pii

S0166-0934(18)30417-8

journal_volume

265

pub_type

杂志文章

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