Defence Institute of Physiology and Allied Sciences (DIPAS), Ministry of Defence, DRDO, Govt. of India, Lucknow Road, Timarpur, Delhi, 110054, India.
Vanguard Diagnostics Private Limited, Okhla Industrial Area, New Delhi, 110020, India.
Appl Microbiol Biotechnol. 2022 Sep;106(18):6225-6238. doi: 10.1007/s00253-022-12113-8. Epub 2022 Aug 17.
The severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) pandemic has adversely affected humankind and caused millions of deaths globally since January 2020. Robust and quick serological tests such as antibody detection assays for SARS-CoV-2 provide relevant information and aid in the process of vaccine development and diagnostics, as well as in sero-epidemiological monitoring of antibody response to the virus. The receptor-binding domain (RBD) of spike and nucleocapsid protein are specific targets for detecting SARS-CoV-2 antibodies. Here, we present the development of a stable spike (S) and nucleocapsid (N) protein-based ELISA antibody detection test "CoroSuchak," with 99% sensitivity, 98% specificity, cost-effective, and detection in a minimum time for serodiagnosis and mass screening of the population for antibodies against SARS-CoV-2. Blood samples were analyzed from 374 SARS-CoV-2 reverse transcription-polymerase chain reaction (RT-PCR) positive, 772 negative and asymptomatic, and 874 random groups of subjects. We found that the antibody titer was significantly higher (p < 0.0001) in infected and vaccinated group compared to the only vaccinated and only infected group. Using enzyme-linked immunosorbent assay (ELISA), we detected SARS-CoV-2 immunoglobulin G (IgG) antibodies in 118/123 (96%) infected individuals, 570/653 (87%) non-infected but vaccinated individuals, 231/237 (97%) individuals who were both infected and vaccinated, and 499/874 (57%) from randomly selected individuals from the first and second waves of the pandemic. Similarly in the third wave, 14/14 (100%) infected and 16/20 (80%) RT-PCR-negative but symptomatic subjects were detected. Thus, the highly sensitive and specific in-house developed ELISA antibody detection kit "CoroSuchak" is extremely useful to determine the seroprevalence of SARS-CoV-2 antibodies in the coronavirus-exposed population. KEY POINTS: •Indigenous kit using a combination of spike and nucleocapsid proteins and peptide sequences. •High sensitivity and specificity to detect variants. •Highly sensitive for mass screening.
严重急性呼吸综合征冠状病毒 2 型（SARS-CoV-2）大流行自 2020 年 1 月以来对人类造成了严重影响，并在全球范围内导致数百万人死亡。快速而稳健的血清学检测方法，如 SARS-CoV-2 的抗体检测，为疫苗开发和诊断以及对病毒抗体反应的血清流行病学监测提供了相关信息和帮助。刺突（S）和核衣壳（N）蛋白的受体结合域（RBD）是检测 SARS-CoV-2 抗体的特异性靶标。在这里，我们提出了一种稳定的基于刺突（S）和核衣壳（N）蛋白的酶联免疫吸附试验（ELISA）抗体检测试剂盒“CoroSuchak”的开发，该试剂盒具有 99%的灵敏度、98%的特异性、具有成本效益，并且可以在最短的时间内用于检测人群对 SARS-CoV-2 的抗体。对 374 名 SARS-CoV-2 逆转录-聚合酶链反应（RT-PCR）阳性、772 名阴性和无症状以及 874 名随机组别的血液样本进行了分析。我们发现，与仅接种疫苗和仅感染组相比，感染和接种疫苗组的抗体滴度显著更高（p<0.0001）。使用酶联免疫吸附试验（ELISA），我们在 123 名（96%）感染个体、653 名（87%）未感染但接种疫苗个体、231 名（97%）既感染又接种疫苗个体和 874 名（57%）随机选择的个体中检测到 SARS-CoV-2 免疫球蛋白 G（IgG）抗体。从大流行的第一波和第二波中选择的个体。同样在第三波中，检测到 14/14（100%）感染和 16/20（80%）RT-PCR 阴性但有症状的个体。因此，这种高度敏感和特异的内部开发的 ELISA 抗体检测试剂盒“CoroSuchak”对于确定冠状病毒暴露人群中 SARS-CoV-2 抗体的血清流行率非常有用。关键点：• 使用刺突和核衣壳蛋白以及肽序列组合的本土试剂盒。• 对变体具有高灵敏度和特异性。• 非常适合大规模筛查。
