A rapid visual detection method for porcine teschovirus through reverse transcription recombinase-aided amplification coupled with lateral flow dipstick.

IF 2.3 2区 农林科学 Q1 VETERINARY SCIENCES
Qiu-Yong Chen, Zhi-Hua Sun, Ru-Jing Chen, Xue-Min Wu, Yong-Liang Che, Ren-Jie Wu, Jing-Li Qiu, Bing He, Long-Bai Wang, Lun-Jiang Zhou
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引用次数: 0

Abstract

Background: Porcine teschovirus (PTV) is an important enteropathogen, associated with symptoms of polioencephalomyelitis, pneumonia, pericarditis, myocarditis, diarrhea, and reproductive disorders in pigs. Rapid and precise diagnostic methods are essential for managing PTV infections. The study introduced a simple, quick, and visual approach for detecting PTV through the use of RT-RAA coupled with LFD.

Results: The procedures of RT-RAA-LFD for PTV could be carried out with 1.0 μmol/L primer concentration and 2.0 μmol/L probe concentration at 37 °C for 20 min, and the amplification result could be visualized within 5 min through LFD detection. Meanwhile, the assay established in this study showed no interaction with other associated diarrhea viruses, and has high specificity to PTV, with a minimal detection limit of 10 copies/μL and good repeatability. 128 clinical samples suspected of having a PTV infection were tested by RT-PCR and RT-RAA-LFD, respectively. The total diagnostic coincidence rate was 98.44% (126/128) with a Kappa value of 0.96(K ≥ 0.75), demonstrating a high degree of agreement to detect PTV for the two methods.

Conclusions: The RT-RAA-LFD assay created in this research displayed quick response, specificity, and sensitivity, capable of successfully detecting PTV in less than 25 min, providing an easy-to-use diagnostic instrument for rapid and visual PTV detection, especially suitable for labs and low-resource environments.

逆转录重组酶辅助扩增与侧流试纸相结合的猪teschovirus快速视觉检测方法。
背景:猪蛲虫病毒(PTV)是一种重要的肠道病原体,与猪脊髓灰质炎、肺炎、心包炎、心肌炎、腹泻和生殖障碍等症状相关。快速、准确的诊断方法对于治疗PTV感染至关重要。该研究介绍了一种简单、快速、直观的方法,通过使用RT-RAA结合LFD来检测PTV。结果:RT-RAA-LFD检测PTV时,引物浓度为1.0 μmol/L,探针浓度为2.0 μmol/L,温度为37℃,反应时间为20 min, LFD检测可在5 min内显示扩增结果。同时,该方法与其他相关腹泻病毒无相互作用,对PTV具有较高的特异性,最低检出限为10拷贝/μL,重复性好。采用RT-PCR和RT-RAA-LFD分别对128例疑似PTV感染的临床样本进行检测。总诊断符合率为98.44% (126/128),Kappa值为0.96(K≥0.75),表明两种方法检测PTV的一致性很高。结论:本研究建立的RT-RAA-LFD检测方法反应速度快,特异性强,灵敏度高,可在25 min内成功检测PTV,为快速、直观地检测PTV提供了一种易于使用的诊断工具,特别适用于实验室和低资源环境。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
BMC Veterinary Research
BMC Veterinary Research VETERINARY SCIENCES-
CiteScore
4.80
自引率
3.80%
发文量
420
审稿时长
3-6 weeks
期刊介绍: BMC Veterinary Research is an open access, peer-reviewed journal that considers articles on all aspects of veterinary science and medicine, including the epidemiology, diagnosis, prevention and treatment of medical conditions of domestic, companion, farm and wild animals, as well as the biomedical processes that underlie their health.
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