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

被引:0
作者
Chen, Qiu-Yong [1 ,2 ]
Sun, Zhi-Hua [1 ,2 ]
Chen, Ru-Jing [1 ,2 ]
Wu, Xue-Min [1 ,2 ]
Che, Yong-Liang [1 ,2 ]
Wu, Ren-Jie [1 ,2 ]
Qiu, Jing-Li [1 ,2 ]
He, Bing [1 ,2 ]
Wang, Long-Bai [1 ,2 ]
Zhou, Lun-Jiang [1 ,2 ]
机构
[1] Fujian Acad Agr Sci, Inst Anim Husb & Vet Med, Fuzhou 350013, Peoples R China
[2] Fujian Anim Dis Control Technol Dev Ctr, Fuzhou 350013, Peoples R China
关键词
Porcine teschovirus; Visual detection method; Recombinase-aided amplification; Lateral flow dipstick; GENETIC DIVERSITY; PIG-POPULATIONS; FECAL SAMPLES; ENTEROVIRUS-G; ASSAY; POLIOENCEPHALOMYELITIS; PICORNAVIRUSES; SAPELOVIRUS; PREVALENCE; VP1;
D O I
10.1186/s12917-024-04442-9
中图分类号
S85 [动物医学(兽医学)];
学科分类号
0906 ;
摘要
BackgroundPorcine 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.ResultsThe procedures of RT-RAA-LFD for PTV could be carried out with 1.0 mu mol/L primer concentration and 2.0 mu mol/L probe concentration at 37 degrees 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/mu L 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.ConclusionsThe 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.
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页数:9
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