Simultaneous detection of novel goose parvovirus and novel duck reovirus by SYBR Green I-based duplex real-time quantitative polymerase chain reaction.

IF 2.6 4区 生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY 3 Biotech Pub Date : 2024-11-01 Epub Date: 2024-11-03 DOI:10.1007/s13205-024-04139-8
Yimin Wang, Yong Wang, Zhuangli Bi, Yuhan Liu, Chunchun Meng, Jie Zhu, Guangqing Liu, Chuanfeng Li
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Abstract

Co-infection with novel goose parvovirus (NGPV) and novel duck reovirus (NDRV) is common, significantly impeding duck growth and resulting in considerable economic losses within the duck farming industry. To facilitate rapid and accurate diagnosis and differentiation of these two viruses, this study developed a SYBR Green I-based duplex real-time quantitative polymerase chain reaction (qPCR) assay. This assay enabled the simultaneous detection of NGPV and NDRV by exploiting their distinct melting temperatures (Tm): 78.5 ± 0.50 °C for NGPV and 84.5 ± 0.50 °C for NDRV. No amplification was observed for other prevalent non-target duck viruses. The intra- and inter-assay coefficients of variation were less than 1.75%. The assay showed good performance with the same detection limit of 102 copies/μL for both NGPV and NDRV. The results of the clinical testing indicated that 45.3% (34/75) of the samples tested positive for NGPV, while 38.7% (29/75) were positive for NDRV. Notably, 13.3% (10/75) exhibited co-infection. These results revealed that the sensitivity of the developed method exceed that of conventional polymerase chain reaction (PCR). The developed method for the identifying of NGPV and NDRV shows good specificity, sensitivity, and repeatability, rendering it an effective tool for the simultaneous detection of co-infection with NGPV and NDRV.

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利用基于 SYBR Green I 的双工实时定量聚合酶链式反应同时检测新型鹅细小病毒和新型鸭细小病毒。
新型鹅副粘病毒(NGPV)和新型鸭再病毒(NDRV)的混合感染很常见,严重阻碍了鸭子的生长,给养鸭业造成了巨大的经济损失。为了快速准确地诊断和区分这两种病毒,本研究开发了一种基于 SYBR Green I 的双工实时定量聚合酶链反应(qPCR)检测方法。通过利用 NGPV 和 NDRV 不同的熔解温度 (Tm),该检测方法可同时检测这两种病毒:NGPV 为 78.5 ± 0.50 °C,NDRV 为 84.5 ± 0.50 °C。其他流行的非目标鸭病毒未发现扩增。测定内和测定间的变异系数均小于 1.75%。该检测方法性能良好,对 NGPV 和 NDRV 的检测限均为 102 拷贝/μL。临床检测结果表明,45.3%(34/75)的样本对 NGPV 检测呈阳性,38.7%(29/75)的样本对 NDRV 检测呈阳性。值得注意的是,13.3%(10/75)的样本表现出合并感染。这些结果表明,所开发方法的灵敏度超过了传统的聚合酶链反应(PCR)。所开发的鉴定 NGPV 和 NDRV 的方法具有良好的特异性、灵敏度和重复性,是同时检测 NGPV 和 NDRV 共同感染的有效工具。
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来源期刊
3 Biotech
3 Biotech Agricultural and Biological Sciences-Agricultural and Biological Sciences (miscellaneous)
CiteScore
6.00
自引率
0.00%
发文量
314
期刊介绍: 3 Biotech publishes the results of the latest research related to the study and application of biotechnology to: - Medicine and Biomedical Sciences - Agriculture - The Environment The focus on these three technology sectors recognizes that complete Biotechnology applications often require a combination of techniques. 3 Biotech not only presents the latest developments in biotechnology but also addresses the problems and benefits of integrating a variety of techniques for a particular application. 3 Biotech will appeal to scientists and engineers in both academia and industry focused on the safe and efficient application of Biotechnology to Medicine, Agriculture and the Environment.
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