Tolah Ahmed Majdi K, Sohrab Sayed S, Tolah Khaled Majdi K, Hassan Ahmed M, El-Kafrawy Sherif A, Azhar Esam I
Special Infectious Agents Unit, King Fahd Medical Research Center, King Abdulaziz University, P.O. Box 128442, Jeddah 21362, Saudi Arabia.
Department of Medical Laboratory Technology, Faculty of Applied Medical Sciences, King Abdulaziz University, P.O. Box 80324, Jeddah 21589, Saudi Arabia.
Diagnostics (Basel). 2021 May 30;11(6):994. doi: 10.3390/diagnostics11060994.
The unusual cases of pneumonia outbreak were reported from Wuhan city in late December 2019. Serological testing provides a powerful tool for the identification of prior infection and for epidemiological studies. Pseudotype virus neutralization assays are widely used for many viruses and applications in the fields of serology. The accuracy of pseudotype neutralizing assay allows for its use in low biosafety lab and provides a safe and effective alternative to the use of wild-type viruses. In this study, we evaluated the performance of this assay compared to the standard microneutralization assay as a reference. The lentiviral pseudotype particles were generated harboring the Spike gene of SARS-CoV-2. The generated pseudotype particles assay was used to evaluate the activity of neutralizing antibodies in 300 human serum samples from a COVID-19 sero-epidemiological study. Testing of these samples resulted in 55 positive samples and 245 negative samples by pseudotype viral particles assay while microneutralization assay resulted in 64 positive and 236 negative by MN assay. Compared to the MN, the pseudotyped viral particles assay showed a sensitivity of 85.94% and a specificity of 100%. Based on the data generated from this study, the pseudotype-based neutralization assay showed a reliable performance for the detection of neutralizing antibodies against SARS-CoV-2 and can be used safely and efficiently as a diagnostic tool in a biosafety level 2 laboratory.
2019年12月下旬，武汉市报告了几起不同寻常的肺炎疫情。血清学检测为识别既往感染和开展流行病学研究提供了有力工具。假型病毒中和试验广泛应用于多种病毒及血清学领域。假型中和试验的准确性使其可在低生物安全实验室使用，为野生型病毒的使用提供了一种安全有效的替代方法。在本研究中，我们将该试验的性能与作为参考的标准微量中和试验进行了比较评估。构建了携带严重急性呼吸综合征冠状病毒2（SARS-CoV-2）刺突基因的慢病毒假型颗粒。所构建的假型颗粒试验用于评估一项新型冠状病毒肺炎血清流行病学研究中300份人血清样本的中和抗体活性。通过假型病毒颗粒试验检测这些样本，结果为55份阳性样本和245份阴性样本，而微量中和试验通过中和试验得出64份阳性和236份阴性。与中和试验相比，假型病毒颗粒试验的灵敏度为85.94%，特异性为100%。基于本研究产生的数据，基于假型的中和试验在检测针对SARS-CoV-2的中和抗体方面表现出可靠的性能，并且可以作为生物安全2级实验室的诊断工具安全有效地使用。
