Fabiani Marco, Margiotti Katia, Sabatino Manuela, Viola Antonella, Mesoraca Alvaro, Giorlandino Claudio
ALTAMEDICA, Human Genetics, Viale Liegi 45, 00198 Rome, Italy.
Rome Center for Molecular Design, Department of Drug Chemistry and Technology, Sapienza University, Piazzale Aldo Moro 5, 00185 Rome, Italy.
Vaccines (Basel). 2022 Mar 21;10(3):483. doi: 10.3390/vaccines10030483.
Since 2020, the COVID-19 pandemic has spread worldwide, causing health, economic, and social distress. Containment strategies rely on rapid and consistent methodology for molecular detection and characterization. Emerging variants of concern (VOCs) are currently associated with increased infectivity and immune escape (natural defence mechanisms and vaccine). Several VOCs have been detected, including Alpha variant (B.1.1.7), Beta variant (B.1.351), Gamma variant (P.1/B.1.1.28.1) and Delta variant (B.1.617.2), first identified in the UK, South Africa, Brazil and India, respectively. Here, a rapid and low-cost technique was validated to distinguish the Alpha, Beta, Gamma, and Delta SARS-CoV-2 variants by detecting spike gene mutations using a real-time reverse transcription polymerase chain reaction methodology (RT-PCR). A total of 132 positive patients affected by coronavirus disease-19 (COVID-19) were analysed by employing RT-PCR to target single-nucleotide polymorphisms (SNPs) to screen spike protein mutations. All data were validated by the next-generation sequencing (NGS) methodology and using sequences from a public database. Among 132 COVID-19-positive samples, we were able to discriminate all of the investigated SARS-CoV-2 variants with 100% concordance when compared with the NGS method. RT-PCR -based assays for identifying circulating VOCs of SARS-CoV-2 resulted in a rapid method used to identify specific SARS-CoV-2 variants, allowing for a better survey of the spread of the virus and its transmissibility in the pandemic phase.
自2020年以来，新冠疫情已在全球蔓延，造成健康、经济和社会困境。防控策略依赖于用于分子检测和特征分析的快速且一致的方法。目前，令人担忧的新型变体（VOCs）与传染性增加和免疫逃逸（自然防御机制和疫苗）有关。已检测到几种VOCs，包括分别在英国、南非、巴西和印度首次发现的阿尔法变体（B.1.1.7）、贝塔变体（B.1.351）、伽马变体（P.1/B.1.1.28.1）和德尔塔变体（B.1.617.2）。在此，一种快速且低成本的技术通过使用实时逆转录聚合酶链反应方法（RT-PCR）检测刺突基因突变，被验证可区分阿尔法、贝塔伽马和德尔塔SARS-CoV-2变体。通过采用RT-PCR靶向单核苷酸多态性（SNP）来筛查刺突蛋白突变，对总共132名感染新冠病毒病（COVID-19）的阳性患者进行了分析。所有数据均通过下一代测序（NGS）方法并使用公共数据库中的序列进行了验证。在132个COVID-19阳性样本中，与NGS方法相比，我们能够以100%的一致性区分所有研究的SARS-CoV-2变体。基于RT-PCR的检测SARS-CoV-2循环VOCs的方法产生了一种用于识别特定SARS-CoV-2变体的快速方法，有助于在疫情阶段更好地监测病毒的传播及其传染性。
