Soltani-Zangbar Mohammad Sadegh, Aghebati-Maleki Leili, Hajivalili Mahsa, Haji-Fatahaliha Mostafa, Motavalli Roza, Mahmoodpoor Ata, Kafil Hossein Samadi, Farhang Sara, Pourakbari Ramin, Jadidi-Niaragh Farhad, Roshangar Leila, Heris Javad Ahmadian, Kamrani Amin, Siahmansouri Homayoon, Hosseini Maryam, Miahipour Abolfazl, Shareghi-Oskoue Oldouz, Parhizkar Forough, Yousefi Mehdi
Stem Cell Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
Department of Immunology, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran.
Gene Rep. 2021 Jun;23:101140. doi: 10.1016/j.genrep.2021.101140. Epub 2021 Apr 13.
As the daily number of coronavirus infection disease 19 (COVID19) patients increases, the necessity of early diagnosis becomes more obvious. In this respect, we aimed to develop a serological test for specifically detecting anti-SARS-CoV2 antibodies.
We collected serum and saliva samples from 609 individuals who work at TBZMED affiliated hospitals in Tabriz, Iran, from April to June of 2020. Real-time PCR technique was used to detect SARS-CoV-2 genome using specific primers. An enzyme linked immunosorbent assay (ELISA) test was designed based on virus nucleocapsid (N), spike (S) and its receptor binding domain (RBD) protein, and the collected sera were subjected to IgM and/or IgG analysis.
Real-time PCR results showed that 66 people were infected with the SARS-CoV-2. Our designed ELISA kit showed 93.75% and 98% of sensitivity and specificity, respectively. In this study, 5.74% of participants had specific IgG against RBD, whereas the percentage for IgM positive individuals was 5.58%. Approximately the same results were observed for S protein. The number of positive participants for NP increased further, and the results of this antigen showed 7.38% for IgG and 7.06% for IgM.
The ELISA test beside real-time PCR could provide a reliable serologic profile for the status of the disease progress and early detection of individuals. More importantly, it possesses the potential to identify the best candidates for plasma donation according to the antibody titers.
随着新型冠状病毒肺炎（COVID-19）患者每日数量的增加，早期诊断的必要性愈发明显。在这方面，我们旨在开发一种专门检测抗SARS-CoV-2抗体的血清学检测方法。
我们于2020年4月至6月从伊朗大不里士市TBZMED附属医院工作的609名个体中收集了血清和唾液样本。使用实时荧光定量PCR技术，通过特异性引物检测SARS-CoV-2基因组。基于病毒核衣壳（N）、刺突（S）及其受体结合域（RBD）蛋白设计了一种酶联免疫吸附测定（ELISA）试验，并对收集的血清进行IgM和/或IgG分析。
实时荧光定量PCR结果显示，66人感染了SARS-CoV-2。我们设计的ELISA试剂盒的灵敏度和特异性分别为93.75%和98%。在本研究中，5.74%的参与者具有针对RBD的特异性IgG，而IgM阳性个体的比例为5.58%。S蛋白的结果大致相同。NP阳性参与者的数量进一步增加，该抗原的结果显示IgG为7.38%，IgM为7.06%。
除实时荧光定量PCR外，ELISA检测可为疾病进展状况和个体的早期检测提供可靠的血清学特征。更重要的是，它有潜力根据抗体滴度识别最佳的血浆捐献候选人。
