Evaluation of an incubation instrument-free reverse transcription recombinase polymerase amplification assay for rapid and point-of-need detection of canine distemper virus.
Evaluation of an incubation instrument-free reverse transcription recombinase polymerase amplification assay for rapid and point-of-need detection of canine distemper virus.
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DOI：
10.1016/j.jviromet.2018.07.007
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发表时间：
2018-10
影响因子：
3.1
通讯作者：
Yuan W
中科院分区：
文献类型：
--
作者：
Wang J;Wang J;Li R;Shi R;Liu L;Yuan W
建立了一种直观、快速的CDV分子检测方法。检测是基于RT-RPA和使用横向流动带来可视化产品。检测在紧握的拳头中进行，体热15 分钟。检测分析的灵敏度和特异度与实时荧光RT-PCR相似。该方法具有简便、快速、可靠的特点。犬瘟热是由犬瘟热病毒(CDV)引起的一种高度传染性和致死性的全身性疾病，在世界范围内自由生活和圈养的食肉动物中。准确、快速、简便地检测CDV对于改进疾病管理和预防暴发至关重要。在本研究中，建立了一种无需孵育仪器的可视化逆转录重组酶聚合酶扩增结合侧向流动试纸条(LFS RT-RPA)的方法，利用针对N蛋白基因的特异的引物和侧向流动(LF)探针检测CDV。CDV LFS RT-RPA检测在闭合拳头中进行，体热15 ，5 min内肉眼可见。该方法可检测出CDV，且与检测的其他病毒无交叉反应。以体外转录的CDVRNA为模板，每次反应的分析灵敏度为9.4 × 10 1拷贝，与实时荧光定量RT-PCR法的结果一致。对32例鼻咽拭子标本进行检测，LFS RT-RPA检测CDV RNA阳性率为62.0%(20/32)，与实时荧光RT-PCR检测结果一致。LFS RT-RPA的检测性能与实时RT-PCR相当，而LFS RT-RPA的检测速度更快、更容易进行。这种新型的CDV LFS RT-RPA方法为在设备简陋的实验室和点播设备中快速、可靠地检测CDV提供了一种有吸引力和前景的工具，对于低资源环境下的CD控制具有重要意义。
Visual, rapid molecular assay for detection of CDV was developed. Assay is based on RT-RPA and use of a lateral flow strip to visualize product. Assay was performed in closed fists using body heat for 15 min. Assay analytical sensitivity and specificity was similar to a real-time RT-PCR. Assay was demonstrated to be simple, convenient, rapid and reliable for detection of CDV. Canine distemper, caused by Canine distemper virus (CDV), is a highly contagious and fatal systemic disease in free-living and captive carnivores worldwide. Accurate, rapid and simple detection of CDV is critical to improve disease management and prevent outbreaks. In this study, a visible and incubation instrument-free reverse-transcription recombinase polymerase amplification assay combined with lateral flow strip (LFS RT-RPA) was developed to detect CDV using primers and lateral flow (LF) probe specific for the nucleocapsid (N) protein gene. The CDV LFS RT-RPA assay was performed in a closed fist using body heat for 15 min, and the products were visible to the naked eyes on the LFS within 5 min. The assay could detect CDV, and there was no cross-reaction with the other viruses tested. Using the in vitro transcribed CDV RNA as template, the analytical sensitivity was 9.4 × 101 copies per reaction, which was the same result as that of a real-time RT-PCR. The assay performance was further evaluated by testing 32 nasal/oropharyngeal swab samples, and CDV RNA positive rate was 62.0% (20/32) by LFS RT-RPA, which was the same result as that of the real-time RT-PCR assay. The performance of the LFS RT-RPA was comparable to real-time RT-PCR, while the LFS RT-RPA assay was much faster and easier to perform. The novel CDV LFS RT-RPA assay provides an attractive and promising tool for rapid and reliable detection of CDV in the underequipped laboratory and point-of-need facility, which is of great significance in CD control in low resource settings.
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影响因子：
3.1
作者：
Elia G;Camero M;Losurdo M;Lucente MS;Larocca V;Martella V;Decaro N;Buonavoglia C
通讯作者：
Buonavoglia C
影响因子：
4.8
作者：
Tan B;Wen YJ;Wang FX;Zhang SQ;Wang XD;Hu JX;Shi XC;Yang BC;Chen LZ;Cheng SP;Wu H
通讯作者：
Wu H
影响因子：
5.4
作者：
Seki, F;Ono, N;Yanagi, Y
通讯作者：
Yanagi, Y
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
影响因子：
3.1
作者：
Elia, Gabriella;Decaro, Nicola;Buonavoglia, Canio
通讯作者：
Buonavoglia, Canio