Ruano-Gallego David, García-Villadangos Miriam, Moreno-Paz Mercedes, Gómez-Elvira Javier, Postigo Marina, Simón-Sacristán María, Reyburn Hugh T, Carolis Carlo, Rodrigo Natalia, Codeseira Yaiza B, Rueda Paloma, Zúñiga Sonia, Enjuanes Luis, Parro Victor
Department of Molecular Evolution, Centro de Astrobiología (INTA-CSIC), Carretera de Ajalvir km 4, Torrejón de Ardoz, Madrid, 28850, Spain.
Space Payload Department, Instituto Nacional de Técnica Aeroespacial (INTA), Carretera de Ajalvir km 4, Torrejón de Ardoz, Madrid, 28850, Spain.
Microb Biotechnol. 2021 May;14(3):1228-1236. doi: 10.1111/1751-7915.13801.
The surge of SARS-CoV-2 has challenged health systems worldwide and efficient tests to detect viral particles, as well as antibodies generated against them, are needed. Specificity, sensitivity, promptness or scalability are the main parameters to estimate the final performance, but rarely all of them match in a single test. We have developed SCOVAM, a protein microarray with several viral antigens (spike, nucleocapsid, main protease Nsp5) as capturing probes in a fluorescence immunoassay for COVID-19 serological testing. SCOVAM depicts IgG and IgM antibody responses against each of these proteins of 22 individuals in a single microscope slide. It detects specific IgM (0.094 μg ml ) and IgG (~0.017 μg ml ) and is scalable and cost-effective. We validated SCOVAM by comparing with a widely used chemiluminescent commercial serological test (n = 742). SCOVAM showed twice the sensitivity and allowed following seroconversion in a single assay. By analysing the prevalence 4 months later in a subset of 76 positive sera, we still detected 93.42% of positives, almost doubling the detection of the commercial assay. The higher sensitivity of SCOVAM is especially relevant to screen sera for convalescent plasma-based treatments, high-throughput antibody response monitoring after vaccination or evaluation of vaccine efficiency.
严重急性呼吸综合征冠状病毒2(SARS-CoV-2)的激增给全球卫生系统带来了挑战,因此需要有效的检测方法来检测病毒颗粒以及针对它们产生的抗体。特异性、敏感性、及时性或可扩展性是评估最终性能的主要参数,但很少有一项检测能同时满足所有这些参数。我们开发了SCOVAM,这是一种蛋白质微阵列,在用于COVID-19血清学检测的荧光免疫分析中,将几种病毒抗原(刺突蛋白、核衣壳蛋白、主要蛋白酶Nsp5)作为捕获探针。SCOVAM能在一张显微镜载玻片上描绘出22名个体针对这些蛋白质中每一种的IgG和IgM抗体反应。它能检测特异性IgM(0.094μg/ml)和IgG(约0.017μg/ml),并且具有可扩展性和成本效益。我们通过与一种广泛使用的化学发光商业血清学检测方法(n = 742)进行比较,对SCOVAM进行了验证。SCOVAM的灵敏度提高了两倍,并且能够在一次检测中跟踪血清转化情况。通过分析4个月后76份阳性血清亚组中的流行率,我们仍检测到93.42%的阳性结果,几乎是商业检测方法检测率的两倍。SCOVAM较高的灵敏度对于筛选用于基于康复血浆的治疗的血清、疫苗接种后高通量抗体反应监测或疫苗效率评估尤为重要。