Development and validation of a multiplex RT-qPCR method for the simultaneous detection of influenza type A, B and SARS-COV-2 viruses

Q3 Medicine

Medicine in Novel Technology and Devices Pub Date : 2025-01-22 DOI:10.1016/j.medntd.2025.100350

Samira Karimkhani , Ehsan Lotfi , Fatemeh Karamali , Mahsa DarestaniFarahani , Reza Keikha , Mahmood Barati

{"title":"Development and validation of a multiplex RT-qPCR method for the simultaneous detection of influenza type A, B and SARS-COV-2 viruses","authors":"Samira Karimkhani , Ehsan Lotfi , Fatemeh Karamali , Mahsa DarestaniFarahani , Reza Keikha , Mahmood Barati","doi":"10.1016/j.medntd.2025.100350","DOIUrl":null,"url":null,"abstract":"<div><div>The study aimed to evaluate the diagnosis between SARS-COV-2 and influenza viruses using the Multiplex qPCR molecular method, highlighting the importance of these methods in disease management.</div><div>In this study, primers and probes were designed for the hemagglutinin region (HA) of influenza A, the M region of influenza B virus, and the RdRp region of SARS-COV-2. Optimization was performed using qPCR methods, and the method's analytical sensitivity and specificity were assessed. Finally, the process was compared to the commercial kit, Generi-Biotech Company (GB SARS-CoV-2 Influenza A/B).</div><div>The best annealing temperature for this method was determined to be 58 °C. Analytical sensitivity showed detection limits of 500 copies of the virus genome for SARS-CoV-2, 250 copies for influenza A, and 500 copies for influenza B. Clinical evaluations confirmed that the designed kit exhibited 100 % sensitivity and specificity, identical to the GB commercial kit, establishing its comparable diagnostic performance.</div></div>","PeriodicalId":33783,"journal":{"name":"Medicine in Novel Technology and Devices","volume":"25 ","pages":"Article 100350"},"PeriodicalIF":0.0000,"publicationDate":"2025-01-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Medicine in Novel Technology and Devices","FirstCategoryId":"1085","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S2590093525000013","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"Medicine","Score":null,"Total":0}

引用次数: 0

Abstract

The study aimed to evaluate the diagnosis between SARS-COV-2 and influenza viruses using the Multiplex qPCR molecular method, highlighting the importance of these methods in disease management.

In this study, primers and probes were designed for the hemagglutinin region (HA) of influenza A, the M region of influenza B virus, and the RdRp region of SARS-COV-2. Optimization was performed using qPCR methods, and the method's analytical sensitivity and specificity were assessed. Finally, the process was compared to the commercial kit, Generi-Biotech Company (GB SARS-CoV-2 Influenza A/B).

The best annealing temperature for this method was determined to be 58 °C. Analytical sensitivity showed detection limits of 500 copies of the virus genome for SARS-CoV-2, 250 copies for influenza A, and 500 copies for influenza B. Clinical evaluations confirmed that the designed kit exhibited 100 % sensitivity and specificity, identical to the GB commercial kit, establishing its comparable diagnostic performance.

Abstract Image

查看原文本刊更多论文

同时检测甲型流感、乙型流感和SARS-COV-2病毒的多重RT-qPCR方法的建立和验证

本研究旨在评价利用多重qPCR分子方法对SARS-COV-2和流感病毒的诊断，强调这些方法在疾病管理中的重要性。本研究针对甲型流感病毒的血凝素区（HA）、乙型流感病毒的M区和SARS-COV-2的RdRp区设计了引物和探针。采用qPCR方法进行优化，并对方法的分析灵敏度和特异性进行评价。最后，将该过程与通用生物技术公司（GB SARS-CoV-2 Influenza A/B）的商用试剂盒进行比较。该方法的最佳退火温度为58℃。分析灵敏度显示，SARS-CoV-2病毒基因组的检出限为500个拷贝，甲型流感为250个拷贝，乙型流感为500个拷贝。临床评价证实，设计的试剂盒具有100%的灵敏度和特异性，与GB商用试剂盒相同，建立了可比的诊断性能。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

求助全文

约1分钟内获得全文求助全文

来源期刊