Development of recombinase polymerase amplification assays for rapid and visual detection of canine distemper virus infecting giant panda.
Development of recombinase polymerase amplification assays for rapid and visual detection of canine distemper virus infecting giant panda.
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开发重组酶聚合酶扩增检测方法，用于快速、直观地检测感染大熊猫的犬瘟热病毒
DOI：
10.1186/s12917-021-02880-3
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发表时间：
2021-04-23
影响因子：
2.6
通讯作者：
Feng N
中科院分区：
文献类型：
--
作者：
Huang P;Yu Y;Meng X;Wang T;Yan F;Li E;Shi Z;He H;Yang S;Xia X;Wang J;Feng N
犬瘟热病毒(CDV)是一种囊膜负链RNA病毒，变异率高，宿主范围不断扩大。值得注意的是，CDV从犬类体内的病毒库溢出，感染了大熊猫。众所周知，大熊猫，特别是圈养大熊猫，容易自然感染CDV。CDV的高致死率对大熊猫种群的安全构成了严重威胁。然而，到目前为止，针对大熊猫犬瘟的疫苗或药物还没有开发出来。因此，能够实现准确现场检测的CDV快速检测，对于及时落实防控措施具有重要意义。在这项研究中，我们建立了针对CDV N基因的核酸可视化方法，将逆转录重组酶聚合酶扩增和封闭垂直流动可视化条带(RT-RPA-VF)相结合。RT-RPA-VF不需要复杂的设备，在35 °C下30 分钟可快速检测，而检测限为5 × 101Copes/μL RNA转录本和100.5 TCID50毫升− 1病毒。结果表明，该方法对CDV具有较高的特异性，与其他感染大熊猫的病毒无交叉反应。与RT-qPCR比较，RT-RPA-VF检测29例临床标本的敏感性和特异性分别为100%和100%。RT-RPA-VF与RT-qPCR的符合率为100%(kappa = 1)，表明RT-RPA-VF对临床标本具有良好的诊断性能。RT-RPA-VF为CDV的简便、敏感和特异性鉴定提供了一种新的选择，并在圈养大熊猫和野生大熊猫的护理点诊断中显示出巨大的潜力。
Canine distemper virus (CDV) is an enveloped negative-strand RNA virus that exhibits a high mutation rate and continuously expands the range of hosts. Notably, CDV has infected giant panda with spill over from viral reservoirs in canines. Giant pandas (Ailuropoda melanoleuca), especially captive pandas, are known to be susceptible to natural infection with CDV. The high fatality rate of CDV poses a serious threat to the safety of the giant panda population. However, vaccines or drugs for canine distemper in giant pandas have not been developed to date. Therefore, a rapid test that can achieve accurate onsite detection of CDV is important to enable the timely implementation of control measures. In this study, we established a nucleic acid visualization assay for targeting the CDV N gene by using combines reverse transcription recombinase polymerase amplification with a closed vertical flow visualization strip (RT-RPA-VF). The RT-RPA-VF assay does not require sophisticated equipment, and it was determined to provide rapid detection at 35 °C for 30 min, while the limit of detection was 5 × 101 copies/μl RNA transcripts and 100.5 TCID50 ml− 1 viruses. The results showed that the assay was high specific to CDV and had no cross-reactivity with other viruses infecting the giant panda. Compared with RT-qPCR, RT-RPA-VF assay had a sensitivity of 100% and a specificity of 100% in 29 clinical samples. The coincidence rate between RT-RPA-VF and RT-qPCR was 100% (kappa = 1), indicating that the RT-RPA-VF assay possessed good diagnostic performance on clinical samples. The RT-RPA-VF provides a novel alternative for the simple, sensitive, and specific identification of CDV and showed great potential for point of care diagnostics for captive and wild giant panda.
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影响因子：
12.3
作者：
Fan, Huizhong;Wu, Qi;Hu, Yibo
通讯作者：
Hu, Yibo
开发重组酶聚合酶扩增结合侧流试纸法检测牛短暂热病毒
影响因子：
3.3
作者：
Hou P;Zhao G;Wang H;He C;Huan Y;He H
通讯作者：
He H
影响因子：
3.1
作者：
Wang J;Wang J;Li R;Shi R;Liu L;Yuan W
通讯作者：
Yuan W
影响因子：
3.7
作者：
Lillis L;Lehman D;Singhal MC;Cantera J;Singleton J;Labarre P;Toyama A;Piepenburg O;Parker M;Wood R;Overbaugh J;Boyle DS
通讯作者：
Boyle DS
DOI：
10.1136/mp.52.3.158
发表时间：
1999-06-01
期刊：
JOURNAL OF CLINICAL PATHOLOGY-MOLECULAR PATHOLOGY
影响因子：
--
作者：
Caldarelli-Stefano, R;Vago, L;Costanzi, G
通讯作者：
Costanzi, G