Valadan Reza, Golchin Soheila, Alizadeh-Navaei Reza, Haghshenas Mohammadreza, Zargari Mehryar, Mousavi Tahoora, Ghamati Mohammad
Molecular and Cell Biology Research Center, Faculty of Medicine, Mazandaran University of Medical Sciences, Sari, Iran.
Department of Immunology, Faculty of Medicine, Mazandaran University of Medical Sciences, Sari, Iran.
AMB Express. 2022 Sep 2;12(1):112. doi: 10.1186/s13568-022-01454-2.
COVID-19 currently is the main cause of the severe acute respiratory disease and fatal outcomes in human beings worldwide. Several genes are used as targets for the detection of SARS-CoV-2, including the RDRP, N, and E genes. The present study aimed to determine the RDRP, N, and E genes expressions of SARS-CoV- 2 in clinical samples. For this purpose, 100 SARS-CoV-2 positive samples were collected from diagnostic laboratories of Mazandaran province, Iran. After RNA extraction, the real-time reverse transcription PCR (real-time RT-PCR) assay was performed for differential gene expressions' analysis of N, E, and RDRP. The threshold cycle (Ct) values for N, RDRP, and E targets of 100 clinical samples for identifying SARS-CoV-2 were then evaluated using quantitative real-time PCR (qRT-PCR). This result suggests N gene as a potential target for the detection of the SARS-CoV-2, since it was observed to be highly expressed in the nasopharyngeal or oropharynges of COVID-19 patients (P < 0.0001). Herein, we showed that SARS-CoV- 2 genes were differentially expressed in the host cells. Therefore, to reduce obtaining false negative results and to increase the sensitivity of the available diagnostic tests, the target genes should be carefully selected based on the most expressed genes in the cells.
新型冠状病毒肺炎(COVID-19)目前是全球人类严重急性呼吸道疾病和致死的主要原因。几个基因被用作检测严重急性呼吸综合征冠状病毒2(SARS-CoV-2)的靶点,包括RNA依赖性RNA聚合酶(RDRP)、核衣壳蛋白(N)和包膜蛋白(E)基因。本研究旨在确定临床样本中SARS-CoV-2的RDRP、N和E基因表达情况。为此,从伊朗马赞德兰省的诊断实验室收集了100份SARS-CoV-2阳性样本。RNA提取后,进行实时逆转录聚合酶链反应(实时RT-PCR)检测,以分析N、E和RDRP的差异基因表达。然后使用定量实时聚合酶链反应(qRT-PCR)评估100份临床样本中用于鉴定SARS-CoV-2的N、RDRP和E靶点的阈值循环(Ct)值。该结果表明N基因是检测SARS-CoV-2的潜在靶点,因为在新型冠状病毒肺炎患者的鼻咽或口咽中观察到其高表达(P < 0.0001)。在此,我们表明SARS-CoV-2基因在宿主细胞中差异表达。因此,为了减少出现假阴性结果并提高现有诊断检测的灵敏度,应根据细胞中表达最多的基因仔细选择靶基因。