Development of an immunochromatographic strip for serological diagnosis of Porcine hemagglutinating encephalomyelitis virus
Development of an immunochromatographic strip for serological diagnosis of Porcine hemagglutinating encephalomyelitis virus
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猪血凝性脑脊髓炎病毒血清学诊断免疫层析试纸条的研制
DOI：
10.1177/104063871102300214
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发表时间：
2011-03-01
影响因子：
1.5
通讯作者：
Gao, Feng
中科院分区：
文献类型：
--
作者：
Chen, Keyan;He, Wenqi;Gao, Feng
建立了猪血凝性脑脊髓炎病毒（PHEV）抗体的免疫层析检测方法。以胶体金标记的兔抗猪免疫球蛋白G （IgG）作为检测试剂，将PHEV重组抗原和山羊抗兔IgG分别包被在原型试纸和对照线上。免疫层析条能在10 min内特异性检测出血凝抑制（HI）效价为2的PHEV抗体。在室温下保存6个月或在4℃下保存12个月不改变其敏感性和特异性。以HI为参比试验，确定免疫层析条的相对特异性为93.41%，灵敏度为98.42%。免疫层析条检测结果与免疫层析条检测结果吻合较好（kappa = 0.926）。此外，酶联免疫吸附试验与免疫层析条带之间有很强的一致性（kappa = 0.929）。将免疫层析试纸条用于吉林省1117份血清的血清学诊断，血清阳性率从辽源区的6.5%到长春区的81.6%不等。此外，许多仔猪对PHEV血清呈阳性，表明母体抗体可能通过初乳转移。该免疫层析条具有较高的特异性、敏感性和稳定性，可用于PHEV抗体的现场检测，从而监测动物群体中PHEV感染情况。
An immunochromatographic strip was developed for the detection of an antibody against Porcine hemagglutinating encephalomyelitis virus (PHEV). Colloidal gold labeled rabbit anti-pig immunoglobulin G (IgG) was used as the detection reagent, and the PHEV recombinant antigens and goat anti-rabbit IgG were coated on the prototype strip and the control lines, respectively. The immunochromatographic strip was capable of specifically detecting PHEV antibodies in serum with a hemagglutination inhibition (HI) titer 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 and specificity. Using HI as a reference test, the relative specificity and sensitivity of the immunochromatographic strip were determined to be 93.41% and 98.42%, respectively. There was a strong agreement between the results obtained by HI and the immunochromatographic strips (kappa = 0.926). Additionally, there was a strong agreement between enzyme-linked immunosorbent assay and immunochromatographic strips (kappa = 0.929). When the immunochromatographic strip was used for serological diagnosis of 1,117 serum samples in Jilin Province in China, the seropositivity ranged from 6.5% in the Liaoyuan District to 81.6% in the Changchun District. Furthermore, many piglets were seropositive to PHEV, indicating the possible transfer of maternal antibodies via the colostrum. Based on the high specificity, sensitivity, and stability of the immunochromatographic strip, it would be suitable for on-site detection of PHEV antibodies in order to monitor PHEV infections in an animal population.