• 文献检索
  • 文档翻译
  • 深度研究
  • 学术资讯
  • Suppr Zotero 插件Zotero 插件
  • 邀请有礼
  • 套餐&价格
  • 历史记录
应用&插件
Suppr Zotero 插件Zotero 插件浏览器插件Mac 客户端Windows 客户端微信小程序
定价
高级版会员购买积分包购买API积分包
服务
文献检索文档翻译深度研究API 文档MCP 服务
关于我们
关于 Suppr公司介绍联系我们用户协议隐私条款
关注我们

Suppr 超能文献

核心技术专利：CN118964589B侵权必究
粤ICP备2023148730 号-1Suppr @ 2026

文献检索

告别复杂PubMed语法，用中文像聊天一样搜索，搜遍4000万医学文献。AI智能推荐，让科研检索更轻松。

立即免费搜索

文件翻译

保留排版，准确专业，支持PDF/Word/PPT等文件格式，支持 12+语言互译。

免费翻译文档

深度研究

AI帮你快速写综述，25分钟生成高质量综述，智能提取关键信息，辅助科研写作。

立即免费体验

用于检测 SARS-CoV-2 的多重实时 RT-PCR 检测方法的建立。

Development of multiplex real-time RT-PCR assay for the detection of SARS-CoV-2.

机构信息

Department of Genetics Research, Institute for Research and Medical Consultations (IRMC), Imam Abdulrahman Bin Faisal University, Dammam, Saudi Arabia.

Department of Biology, College of Science and Institute for Research and Medical Consultations (IRMC), Imam Abdulrahman Bin Faisal University, Dammam, Saudi Arabia.

出版信息

PLoS One. 2021 Apr 29;16(4):e0250942. doi: 10.1371/journal.pone.0250942. eCollection 2021.

DOI:10.1371/journal.pone.0250942
PMID:33914804
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8084238/
Abstract

The outbreak of the new human coronavirus SARS-CoV-2 (also known as 2019-nCoV) continues to increase globally. The real-time reverse transcription polymerase chain reaction (rRT-PCR) is the most used technique in virus detection. However, possible false-negative and false-positive results produce misleading consequences, making it necessary to improve existing methods. Here, we developed a multiplex rRT-PCR diagnostic method, which targets two viral genes (RdRP and E) and one human gene (RP) simultaneously. The reaction was tested by using pseudoviral RNA and human target mRNA sequences as a template. Also, the protocol was validated by using 14 clinical SARS-CoV-2 positive samples. The results are in good agreement with the CDC authorized Cepheid`s Xpert® Xpress SARS-CoV-2 diagnostic system (100%). Unlike single gene targeting strategies, the current method provides the amplification of two viral regions in the same PCR reaction. Therefore, an accurate SARS-CoV-2 diagnostic assay was provided, which allows testing of 91 samples in 96-well plates in per run. Thanks to this strategy, fast, reliable, and easy-to-use rRT-PCR method is obtained to diagnose SARS-CoV-2.

摘要

新型人类冠状病毒 SARS-CoV-2（也称为 2019-nCoV）的爆发继续在全球范围内增加。实时逆转录聚合酶链反应（rRT-PCR）是最常用于病毒检测的技术。然而，可能出现的假阴性和假阳性结果会产生误导性后果，因此有必要改进现有方法。在这里，我们开发了一种多重 rRT-PCR 诊断方法，该方法同时针对两个病毒基因（RdRP 和 E）和一个人类基因（RP）。该反应使用假病毒 RNA 和人靶 mRNA 序列作为模板进行测试。此外，该方案还通过使用 14 个临床 SARS-CoV-2 阳性样本进行了验证。结果与 CDC 授权的 Cepheid`s Xpert® Xpress SARS-CoV-2 诊断系统（100%）一致。与单基因靶向策略不同，目前的方法在同一 PCR 反应中提供了两个病毒区域的扩增。因此，提供了一种准确的 SARS-CoV-2 诊断检测方法，可在每次运行中对 91 个 96 孔板进行检测。由于采用了这种策略，因此获得了快速、可靠且易于使用的 rRT-PCR 方法来诊断 SARS-CoV-2。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/606d/8084238/58bed3765c21/pone.0250942.g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/606d/8084238/05be09c3e990/pone.0250942.g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/606d/8084238/433a35353491/pone.0250942.g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/606d/8084238/58bed3765c21/pone.0250942.g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/606d/8084238/05be09c3e990/pone.0250942.g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/606d/8084238/433a35353491/pone.0250942.g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/606d/8084238/58bed3765c21/pone.0250942.g003.jpg

相似文献

1
Development of multiplex real-time RT-PCR assay for the detection of SARS-CoV-2.用于检测 SARS-CoV-2 的多重实时 RT-PCR 检测方法的建立。
PLoS One. 2021 Apr 29;16(4):e0250942. doi: 10.1371/journal.pone.0250942. eCollection 2021.
2
Clinical evaluation of a multiplex real-time RT-PCR assay for detection of SARS-CoV-2 in individual and pooled upper respiratory tract samples.用于检测个体和混合上呼吸道样本中 SARS-CoV-2 的多重实时 RT-PCR 检测的临床评估。
Arch Virol. 2021 Sep;166(9):2551-2561. doi: 10.1007/s00705-021-05148-1. Epub 2021 Jul 14.
3
Sensitive detection and quantification of SARS-CoV-2 by multiplex droplet digital RT-PCR.采用多重液滴数字 RT-PCR 法对 SARS-CoV-2 进行灵敏检测和定量。
Eur J Clin Microbiol Infect Dis. 2021 Apr;40(4):807-813. doi: 10.1007/s10096-020-04076-3. Epub 2020 Oct 26.
4
Sequence-specific and multiplex detection of COVID-19 virus (SARS-CoV-2) using proofreading enzyme-mediated probe cleavage coupled with isothermal amplification.基于校对酶介导的探针切割联合等温扩增的 COVID-19 病毒（SARS-CoV-2）序列特异性和多重检测。
Biosens Bioelectron. 2021 Apr 15;178:113041. doi: 10.1016/j.bios.2021.113041. Epub 2021 Jan 28.
5
Enhanced throughput of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) real-time RT-PCR panel by assay multiplexing and specimen pooling.通过多重检测和样本混合提高严重急性呼吸综合征冠状病毒 2 （SARS-CoV-2）实时 RT-PCR 检测试剂盒的检测通量。
J Virol Methods. 2021 Jul;293:114149. doi: 10.1016/j.jviromet.2021.114149. Epub 2021 Apr 8.
6
Evaluation of Three Multiplex Real-time Reverse Transcription PCR Assays for Simultaneous Detection of SARS-CoV-2, Influenza A/B, and Respiratory Syncytial Virus in Nasopharyngeal Swabs.评价三种多重实时逆转录聚合酶链反应检测试剂盒在鼻咽拭子标本中同时检测新型冠状病毒、甲型/乙型流感病毒和呼吸道合胞病毒的效果。
J Korean Med Sci. 2021 Dec 13;36(48):e328. doi: 10.3346/jkms.2021.36.e328.
7
Development and performance evaluation of the first in-house multiplex rRT-PCR assay in Bangladesh for highly sensitive detection of SARS-CoV-2.孟加拉国首个内部多重实时 RT-PCR 检测试剂盒的开发和性能评估，用于高灵敏度检测 SARS-CoV-2。
J Virol Methods. 2021 Jul;293:114147. doi: 10.1016/j.jviromet.2021.114147. Epub 2021 Apr 1.
8
Sample-to-Answer and Routine Real-Time RT-PCR: A Comparison of Different Platforms for SARS-CoV-2 Detection.样本到答案与常规实时逆转录聚合酶链反应：SARS-CoV-2检测不同平台的比较
J Mol Diagn. 2021 Jun;23(6):665-670. doi: 10.1016/j.jmoldx.2021.02.010. Epub 2021 Mar 8.
9
Development and Evaluation of AccuPower COVID-19 Multiplex Real-Time RT-PCR Kit and AccuPower SARS-CoV-2 Multiplex Real-Time RT-PCR Kit for SARS-CoV-2 Detection in Sputum, NPS/OPS, Saliva and Pooled Samples.AccuPower COVID-19 多重实时 RT-PCR 试剂盒和 AccuPower SARS-CoV-2 多重实时 RT-PCR 试剂盒的开发和评估，用于痰液、鼻咽拭子/口咽拭子、唾液和混合样本中 SARS-CoV-2 的检测。
PLoS One. 2022 Feb 10;17(2):e0263341. doi: 10.1371/journal.pone.0263341. eCollection 2022.
10
In-house reverse transcriptase polymerase chain reaction for detection of SARS-CoV-2 with increased sensitivity.采用室内逆转录聚合酶链反应提高 SARS-CoV-2 检测的灵敏度。
Sci Rep. 2021 Sep 9;11(1):17878. doi: 10.1038/s41598-021-97502-1.

引用本文的文献

1
Compact multiplex PCR device for HIV-1 and HIV-2 viral load determination from finger-prick whole blood in resource-limited settings.用于在资源有限环境中从指尖全血测定HIV-1和HIV-2病毒载量的紧凑型多重PCR装置。
Biosens Bioelectron. 2025 Mar 1;271:116997. doi: 10.1016/j.bios.2024.116997. Epub 2024 Nov 26.
2
A multiplexed real-time PCR assay for simultaneous quantification of human immunodeficiency virus and Hepatitis B virus for low-and-middle- income countries.用于中低收入国家的同时定量检测人类免疫缺陷病毒和乙型肝炎病毒的多重实时 PCR 检测方法。
J Virol Methods. 2024 Dec;330:115026. doi: 10.1016/j.jviromet.2024.115026. Epub 2024 Sep 2.
3

本文引用的文献

1
Promising methods for detection of novel coronavirus SARS-CoV-2.检测新型冠状病毒SARS-CoV-2的有前景的方法。
View (Beijing). 2020 Mar;1(1):e4. doi: 10.1002/viw2.4. Epub 2020 Mar 23.
2
Sensitive detection and quantification of SARS-CoV-2 in saliva.唾液中 SARS-CoV-2 的敏感检测和定量。
Sci Rep. 2021 Jun 14;11(1):12425. doi: 10.1038/s41598-021-91835-7.
3
Development of a multiplex Loop-Mediated Isothermal Amplification (LAMP) assay for on-site diagnosis of SARS CoV-2.建立一种多重环介导等温扩增（LAMP）检测方法，用于现场诊断 SARS-CoV-2。
Validation and Suitability Assessment of Multiplex Mesoscale Discovery Immunogenicity Assay for Establishing Serological Signatures Using Vaccinated, Non-Vaccinated and Breakthrough SARS-CoV-2 Infected Cases.
用于通过接种疫苗、未接种疫苗和突破性感染 SARS-CoV-2 的病例建立血清学特征的多重中尺度发现免疫原性检测的验证和适用性评估
Vaccines (Basel). 2024 Apr 18;12(4):433. doi: 10.3390/vaccines12040433.
4
Ultrasensitive and fast detection of SARS-CoV-2 using RT-LAMP without pH-dependent dye.使用 RT-LAMP 进行无 pH 依赖性染料的 SARS-CoV-2 的超灵敏和快速检测。
Funct Integr Genomics. 2024 Jan 20;24(1):16. doi: 10.1007/s10142-024-01297-z.
5
Development of a robust TaqMan probe-based one-step multiplex RT-qPCR for simultaneous detection of SARS-CoV-2 and Influenza A/B viruses.建立一种基于 TaqMan 探针的一步法多重实时 RT-qPCR 方法，用于同时检测 SARS-CoV-2 和流感 A/B 病毒。
BMC Microbiol. 2023 Nov 11;23(1):335. doi: 10.1186/s12866-023-03048-9.
6
Comparison of DNA extraction methods for COVID-19 host genetics studies.用于 COVID-19 宿主遗传学研究的 DNA 提取方法比较。
PLoS One. 2023 Oct 30;18(10):e0287551. doi: 10.1371/journal.pone.0287551. eCollection 2023.
7
Despite low viral titer in saliva samples, Sanger-based SARS-CoV-2 spike gene sequencing is highly applicable for the variant identification.尽管唾液样本中的病毒滴度较低，但基于 Sanger 的 SARS-CoV-2 刺突基因测序对于变异体鉴定非常适用。
BMC Med Genomics. 2023 Aug 24;16(1):199. doi: 10.1186/s12920-023-01633-5.
8
A review of current effective COVID-19 testing methods and quality control.当前有效 COVID-19 检测方法及质量控制综述。
Arch Microbiol. 2023 May 17;205(6):239. doi: 10.1007/s00203-023-03579-9.
9
Graphene surface plasmon sensor for ultra-low-level SARS-CoV-2 detection.用于超低水平 SARS-CoV-2 检测的石墨烯表面等离子体传感器。
PLoS One. 2023 Apr 25;18(4):e0284812. doi: 10.1371/journal.pone.0284812. eCollection 2023.
10
Development of loop-mediated isothermal amplification (LAMP) assays using five primers reduces the false-positive rate in COVID-19 diagnosis.开发使用五条引物的环介导等温扩增 (LAMP) 检测方法可降低 COVID-19 诊断中的假阳性率。
Sci Rep. 2023 Mar 28;13(1):5066. doi: 10.1038/s41598-023-31760-z.
PLoS One. 2021 Mar 3;16(3):e0248042. doi: 10.1371/journal.pone.0248042. eCollection 2021.
4
Novel dual multiplex real-time RT-PCR assays for the rapid detection of SARS-CoV-2, influenza A/B, and respiratory syncytial virus using the BD MAX open system.新型双重多重实时 RT-PCR 检测方法可快速检测 SARS-CoV-2、流感 A/B 和呼吸道合胞病毒，使用的是 BD MAX 开放式系统。
Emerg Microbes Infect. 2021 Dec;10(1):161-166. doi: 10.1080/22221751.2021.1873073.
5
A sensitive and affordable multiplex RT-qPCR assay for SARS-CoV-2 detection.一种用于 SARS-CoV-2 检测的灵敏且经济的多重 RT-qPCR assay。
PLoS Biol. 2020 Dec 15;18(12):e3001030. doi: 10.1371/journal.pbio.3001030. eCollection 2020 Dec.
6
An Overview on SARS-CoV-2 (COVID-19) and Other Human Coronaviruses and Their Detection Capability via Amplification Assay, Chemical Sensing, Biosensing, Immunosensing, and Clinical Assays.严重急性呼吸综合征冠状病毒2（COVID-19）及其他人类冠状病毒概述及其通过扩增检测、化学传感、生物传感、免疫传感和临床检测的检测能力
Nanomicro Lett. 2021;13(1):18. doi: 10.1007/s40820-020-00533-y. Epub 2020 Nov 2.
7
Detection of SARS-CoV-2 RNA by multiplex RT-qPCR.多重 RT-qPCR 检测 SARS-CoV-2 RNA。
PLoS Biol. 2020 Oct 7;18(10):e3000867. doi: 10.1371/journal.pbio.3000867. eCollection 2020 Oct.
8
Detection of the SARS-CoV-2 humanized antibody with paper-based ELISA.基于纸基 ELISA 的 SARS-CoV-2 人源化抗体检测。
Analyst. 2020 Nov 23;145(23):7680-7686. doi: 10.1039/d0an01609h.
9
Development of a New Multiplex Real-Time RT-PCR Assay for Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) Detection.开发一种用于严重急性呼吸综合征冠状病毒 2（SARS-CoV-2）检测的新型多重实时 RT-PCR 检测方法。
J Mol Diagn. 2020 Dec;22(12):1367-1372. doi: 10.1016/j.jmoldx.2020.09.004. Epub 2020 Sep 19.
10
Automated SARS-COV-2 RNA extraction from patient nasopharyngeal samples using a modified DNA extraction kit for high throughput testing.使用改良的DNA提取试剂盒从患者鼻咽样本中自动提取SARS-CoV-2 RNA，用于高通量检测。
Ann Saudi Med. 2020 Sep-Oct;40(5):373-381. doi: 10.5144/0256-4947.2020.373. Epub 2020 Oct 1.