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用于检测牛呼吸道合胞体病毒的纳米颗粒辅助PCR检测方法的开发。

Development of a nanoparticle-assisted PCR assay for detection of bovine respiratory syncytial virus.

作者信息

Liu Zhankui, Li Jianyou, Liu Zeyu, Li Jiawei, Li Zhijie, Wang Chao, Wang Jianke, Guo Li

机构信息

Institute of Special Animal and Plant Sciences, Chinese Academy of Agricultural Sciences, Changchun, 130112, China.

Graduate School of Northeast Agricultural University, Harbin, China.

出版信息

BMC Vet Res. 2019 Apr 11;15(1):110. doi: 10.1186/s12917-019-1858-0.

Abstract

BACKGROUND

Bovine respiratory syncytial virus (BRSV) is a common pathogen causing respiratory disease in cattle and a significant contributor to the bovine respiratory disease (BRD) complex. BRSV is widely distributed around the world, causing severe economic losses. This study we established a new molecular detection method of BRSV pathogen NanoPCR attributed to the combination of nano-particles in traditional PCR (Polymerase chain reaction) technology.

RESULTS

In this study, the BRSV NanoPCR assay was developed, and its specificity and sensitivity were investigated. The results showed that no cross-reactivity was observed for the NanoPCR assay for related viruses, including the infectious bovine rhinotracheitis virus (IBRV), bovine viral diarrhea virus (BVDV), and bovine parainfluenza virus type 3 (BPIV3), and the assay was more sensitive than the conventional PCR assay, with a detection limit of 1.43 × 10 copies recombinant plasmids per reaction, compared with 1.43 × 10 copies for conventional PCR analysis. Moreover, thirty-nine clinical bovine samples collected from two provinces in North-Eastern China, 46.15% were determined BRSV positive by our NanoPCR assay, compared with 23.07% for conventional PCR.

CONCLUSIONS

This is the first report to demonstrate the application of a NanoPCR assay for the detection of BRSV. The sensitive and specific NanoPCR assay developed in this study can be applied widely in clinical diagnosis and field surveillance of BRSV infection.

摘要

背景

牛呼吸道合胞病毒(BRSV)是引起牛呼吸道疾病的常见病原体,也是牛呼吸道疾病(BRD)综合征的重要病因。BRSV在全球广泛分布,造成严重经济损失。本研究通过将纳米颗粒与传统聚合酶链反应(PCR)技术相结合,建立了一种新的BRSV病原体纳米PCR分子检测方法。

结果

本研究开发了BRSV纳米PCR检测方法,并对其特异性和敏感性进行了研究。结果表明,该纳米PCR检测方法对相关病毒,包括传染性牛鼻气管炎病毒(IBRV)、牛病毒性腹泻病毒(BVDV)和牛副流感病毒3型(BPIV3)均无交叉反应,且该检测方法比传统PCR检测方法更敏感,每个反应的检测限为1.43×10拷贝重组质粒,而传统PCR分析的检测限为1.43×10拷贝。此外,从中国东北两个省份采集的39份临床牛样本中,46.1%经本纳米PCR检测法判定为BRSV阳性,而传统PCR检测法的阳性率为23.07%。

结论

这是首次报道纳米PCR检测方法在BRSV检测中的应用。本研究开发的灵敏且特异的纳米PCR检测方法可广泛应用于BRSV感染的临床诊断和现场监测。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9c27/6458741/0310c2e30fe9/12917_2019_1858_Fig1_HTML.jpg

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