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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

作者： Shi, Jianzhou 1 ; Jin, Qianyue 3 ; Zhang, Xiaozhan 4 ; Zhao, Jinbing 2 ; Li, Na 2 ; Dong, Bingxue 2 ; Yu, Jinran 2 ; Yao, Lunguang 2 ;

作者机构： 1.Shennong Lab, Zhengzhou 450046, Peoples R China

2.Nanyang Normal Univ, Sch Life Sci & Technol, Nanyang 473061, Peoples R China

3.Henan Acad Agr Sci, Key Lab Anim Immunol, Zhengzhou 450002, Peoples R China

4.Henan Univ Anim Husb & Econ, Coll Vet Med, Zhengzhou 450046, Peoples R China

5.Nanyang Normal Univ, Cent Route South North Water Divers Project, Henan Field Observat & Res Stn Headwork Wetland Ec, Nanyang 473061, Peoples R China

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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