SARS-CoV-2 Subgenomic N (sgN) Transcripts in Oro-Nasopharyngeal Swabs Correlate with the Highest Viral Load, as Evaluated by Five Different Molecular Methods.
SARS-CoV-2 Subgenomic N (sgN) Transcripts in Oro-Nasopharyngeal Swabs Correlate with the Highest Viral Load, as Evaluated by Five Different Molecular Methods.
复制标题
DOI：
10.3390/diagnostics11020288
复制
发表时间：
2021-02-12
期刊：
影响因子：
--
通讯作者：
Capoluongo E
中科院分区：
文献类型：
--
作者：
Zollo M;Ferrucci V;Izzo B;Quarantelli F;Domenico CD;Comegna M;Paolillo C;Amato F;Siciliano R;Castaldo G;Capoluongo E
COVID-19大流行迫使诊断实验室将重点放在SARS-CoV-2的早期诊断上。分子测试的阳性结果不能回答有关病毒复制、传播能力和不同临床效果的问题。尽管商业上可用的测定法覆盖了一些靶标，但为了提高这些测定法给出的信息的质量，需要鉴定新的生物标志物。因此，由于亚基因组转录物（sgN和sgE）被认为是病毒活性的标志物，我们评估了这些亚基因组转录物与使用五种不同的商业CE-IVD工具获得的基因组扩增的关系。方法：比较5种CE-IVD试剂盒在20例covid -19阳性患者拭子中检测合成SARS-CoV-2病毒构建体（加入TMB或PBS培养基）和靶标（N、E、RdRp和Orf1ab基因）的能力。采用实时RT-qPCR和数字PCR检测sgN和sgE。结果：当将诊断试剂盒应用于添加TMB或PBS（稀释范围为100 pg至0.1 pg）的目标时，没有一个诊断试剂盒遗漏病毒靶基因。然而，一旦将它们应用于从患者拭子中提取的RNA，无论提取程序如何，重叠率从50%到100%不等。sgN RNA转录物仅在病毒载量较高（Ct≤22.5）的样品中检测到，而sgE在所有Ct范围内。结论：5种试剂盒的性能随检测布局的不同而不同。值得注意的是，sgN转录本的检测与较高的病毒载量相关，因此代表了早期和更严重感染的新标记。
The COVID-19 pandemic has forced diagnostic laboratories to focus on the early diagnostics of SARS-CoV-2. The positivity of a molecular test cannot respond to the question regarding the viral capability to replicate, spread, and give different clinical effects. Despite the fact that some targets are covered by commercially-available assays, the identification of new biomarkers is desired in order to improve the quality of the information given by these assays. Therefore, since the subgenomic transcripts (sgN and sgE) are considered markers of viral activity, we evaluated these subgenomic transcripts in relation to the genomic amplification obtained using five different commercial CE-IVD tools. Methods: Five CE-IVD kits were compared in terms of their capability to detect both synthetic SARS-CoV-2 viral constructs (spiked in TMB or PBS medium) and targets (N, E, RdRp and Orf1ab genes) in twenty COVID-19–positive patients’ swabs. The sgN and sgE were assayed by real-time RT-qPCR and digital PCR. Results: None of the diagnostic kits missed the viral target genes when they were applied to targets spiked in TMB or PBS (at dilutions ranging from 100 pg to 0.1 pg). Nevertheless, once they were applied to RNA extracted from the patients’ swabs, the superimposability ranged from 50% to 100%, regardless of the extraction procedure. The sgN RNA transcript was detected only in samples with a higher viral load (Ct ≤ 22.5), while sgE was within all of the Ct ranges. Conclusions: The five kits show variable performances depending on the assay layout. It is worthy of note that the detection of the sgN transcript is associated with a higher viral load, thus representing a new marker of early and more severe infection.
登录
查看更多内容
影响因子：
9.4
作者：
Lieberman, Joshua A.;Pepper, Gregory;Greninger, Alexander L.
通讯作者：
Greninger, Alexander L.
影响因子：
6.8
作者：
Basso, Daniela;Aita, Ada;Plebani, Mario
通讯作者：
Plebani, Mario
影响因子：
16.6
作者：
Alexandersen S;Chamings A;Bhatta TR
通讯作者：
Bhatta TR
影响因子：
2.8
作者：
Prabhakar PK;Lakhanpal J
通讯作者：
Lakhanpal J
影响因子：
64.8
作者：
Woelfel, Roman;Corman, Victor M.;Wendtner, Clemens
通讯作者：
Wendtner, Clemens