The Development of a Sensitive Droplet Digital Polymerase Chain Reaction Test for Quantitative Detection of Goose Astrovirus
文献类型: 外文期刊
第一作者: Shi, Jianzhou
作者: Shi, Jianzhou;Shi, Jianzhou;Zhao, Jinbing;Li, Na;Dong, Bingxue;Yu, Jinran;Yao, Lunguang;Jin, Qianyue;Zhang, Xiaozhan;Yao, Lunguang
作者机构:
关键词: goose astrovirus; droplet digital PCR; quantitative real-time PCR; ORF2 gene; detection
期刊名称:VIRUSES-BASEL ( 影响因子:4.7; 五年影响因子:4.8 )
ISSN:
年卷期: 2024 年 16 卷 5 期
页码:
收录情况: SCI
摘要: Simple Summary: Goose astrovirus (GAstV) is a novel emerging pathogen that causes significant economic losses in waterfowl farming. In this study, we developed a droplet digital polymerase chain reaction (ddPCR) system for the sensitive and accurate quantification of GAstV using the conserved region of the ORF2 gene. The detection limit of ddPCR was 10 copies/mu L, similar to 28 times greater sensitivity than quantitative real-time PCR (qPCR). The specificity of the test was determined by the failure of amplification of other avian viruses. The ddPCR test showed good repeatability and linearity, and the established ddPCR method had high sensitivity and good specificity to GAstV. Clinical sample test results showed that the positive rate of ddPCR was higher than that of qPCR. A convenient, sensitive, and specific detection method for GAstV in field samples is important in order to effectively control GAstV. (1) Goose astrovirus (GAstV) is a novel emerging pathogen that causes significant economic losses in waterfowl farming. A convenient, sensitive, and specific detection method for GAstV in field samples is important in order to effectively control GAstV. Droplet digital polymerase chain reaction (ddPCR) is a novel, sensitive, good-precision, and absolute quantitation PCR technology which does not require calibration curves. (2) In this study, we developed a ddPCR system for the sensitive and accurate quantification of GAstV using the conserved region of the ORF2 gene. (3) The detection limit of ddPCR was 10 copies/mu L, similar to 28 times greater sensitivity than quantitative real-time PCR (qPCR). The specificity of the test was determined by the failure of amplification of other avian viruses. Both ddPCR and qPCR tests showed good repeatability and linearity, and the established ddPCR method had high sensitivity and good specificity to GAstV. Clinical sample test results showed that the positive rate of ddPCR (88.89%) was higher than that of qPCR (58.33%). (4) As a result, our results suggest that the newly developed ddPCR method might offer improved analytical sensitivity and specificity in its GAstV measurements. The ddPCR could be widely applied in clinical tests for GAstV infections.
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