College of Animal Science and Veterinary Medicine, Jilin University, Changchun, China.
Virol J. 2012 Aug 24;9:172. doi: 10.1186/1743-422X-9-172.
The incidence of PHE among pigs in many countries is on the rise, and it has caused great economic losses to the pig industry. Therefore, the development of a sensitive, specific, and easily-performed assay is crucial for the rapid detection and surveillance of PHE-CoV infection and transmission.
An immunochromatographic strip was developed for the detection of PHE-CoV. The colloidal gold-labeled MAb 4D4 was used as the detection reagent, and the MAb 1E2 and goat anti-mouse IgG coated the strip's test and control lines, respectively. The immunochromatographic strip was capable of specifically detecting PHE-CoV with a HA unit of 2 within 10 min. Storage of the strips at room temperature for 6 months or at 4°C for 12 months did not change their sensitivity or specificity. Using RT-PCR as a reference test, the relative specificity and sensitivity of the immunochromatographic strip were determined to be 100% and 97.78%, respectively. There was an excellent agreement between the results obtained by RT-PCR and the immunochromatographic strips (kappa = 0.976). Additionally, there was a strong agreement between the sandwich enzyme-linked immunosorbent assay (ELISA) and immunochromatographic strips (Kappa = 0.976). When the immunochromatographic strips were used for diagnosing PHE-CoV infection in the Jilin Province, the PHE-CoV-positive rate ranged from 61.54% in the Jilin district to 17.95% in the Songyuan district.
Based on its high specificity, sensitivity, and stability, the immunochromatographic strip would be suitable for on-site detection of PHE-CoV for surveillance and epidemiological purposes.
许多国家猪群中的 PHE 发病率呈上升趋势,这给养猪业造成了巨大的经济损失。因此,开发一种敏感、特异、易于操作的检测方法对于快速检测和监测 PHE-CoV 的感染和传播至关重要。
建立了一种用于检测 PHE-CoV 的免疫层析试纸条。胶体金标记的单抗 4D4 作为检测试剂,单抗 1E2 和羊抗鼠 IgG 分别包被在试纸条的检测线和质控线上。该免疫层析试纸条能够特异性地检测到 HA 单位为 2 的 PHE-CoV,在 10 min 内即可完成检测。室温下储存 6 个月或 4°C 下储存 12 个月均不会改变其敏感性或特异性。以 RT-PCR 为参考试验,免疫层析试纸条的相对特异性和敏感性分别为 100%和 97.78%。RT-PCR 与免疫层析试纸条的检测结果具有极好的一致性(kappa 值为 0.976)。此外,夹心 ELISA 与免疫层析试纸条之间也具有很强的一致性(kappa 值为 0.976)。用该免疫层析试纸条对吉林省的 PHE-CoV 感染进行诊断时,PHE-CoV 的阳性率在吉林地区为 61.54%,在松原地区为 17.95%。
基于其高特异性、敏感性和稳定性,该免疫层析试纸条适合用于 PHE-CoV 的现场检测,可用于监测和流行病学研究。