Key Laboratory of Livestock Infectious Diseases, Ministry of Education, Shenyang Agricultural University, Shenyang, Liaoning Province, China.
Key Laboratory of Ruminant Infectious Disease Prevention and Control (East), Ministry of Agriculture and Rural Affairs, Shenyang, Liaoning Province, P.R. China.
Vet Res Commun. 2024 Nov 25;49(1):34. doi: 10.1007/s11259-024-10603-w.
A recently identified virus, chicken circovirus (ChCV), has been linked to the onset of acute gastroenteritis in chicks, a condition that can have a detrimental impact on the overall health and well-being of chickens in a farming setting. In this study, we developed an enzyme-linked immunosorbent assay (ELISA) method for the indirect detection of antibodies against the chicken circovirus (ChCV) through codon optimization, which effectively expressed the capsid protein of the ChCV and utilized it as an encapsulated antigen following purification. In establishing the ELISA method for detecting antibodies using the purified Cap protein as the antigen, the optimal concentration of the antigen was determined to be 1 µg/mL, the optimal blocking solution was identified as 1% bovine serum albumin, the optimal dilution ratio of the serum to be tested was established to be 1:100, and the dilution ratio of the secondary antibody was determined to be 1:5,000. At these thresholds, the sensitivity of the ELISA method was 94.44%, and the specificity was 100%. The testing of 203 clinical samples yielded a positivity rate of 46.8%, indicating that the virus is endemic in chickens. In conclusion, this study established an ELISA method to detect antibodies against chicken circovirus using recombinant Cap protein as antigen, demonstrating good specificity and sensitivity. This lays the foundation for the development of related kits and the detection of infection and epidemiology of chicken circovirus. Meanwhile, the analysis concluded that chicken circovirus infection is more common, and the prevention and control of this disease should be emphasised and strengthened.
最近发现的一种病毒，鸡圆环病毒（ChCV），与雏鸡急性胃肠炎的发生有关，这种疾病会对养殖环境中鸡的整体健康和福利产生不利影响。在这项研究中，我们通过密码子优化开发了一种酶联免疫吸附测定（ELISA）方法，用于间接检测针对鸡圆环病毒（ChCV）的抗体，该方法有效地表达了 ChCV 的衣壳蛋白，并在纯化后将其用作包被抗原。在建立使用纯化的 Cap 蛋白作为抗原检测抗体的 ELISA 方法时，确定抗原的最佳浓度为 1µg/mL，最佳封闭溶液为 1%牛血清白蛋白，最佳待测试血清稀释比例为 1:100，确定了二抗的稀释比例为 1:5,000。在这些阈值下，ELISA 方法的灵敏度为 94.44%，特异性为 100%。对 203 份临床样本的检测结果显示阳性率为 46.8%，表明该病毒在鸡群中流行。总之，本研究建立了一种使用重组 Cap 蛋白作为抗原检测针对鸡圆环病毒抗体的 ELISA 方法，显示出良好的特异性和敏感性。这为相关试剂盒的开发以及鸡圆环病毒感染和流行病学的检测奠定了基础。同时，分析得出鸡圆环病毒感染较为普遍，应加强对该病的防控。
