国际老年医学杂志2024,Vol.45Issue(2) :135-141.DOI:10.3969/j.issn.1674-7593.2024.02.002

快速定性检测人呼吸道样本中冠状病毒基因芯片的制备方法及其效果评价

Preparation Method of Gene Chip for Rapid Qualitative Detection of Coronavirus in Human Respiratory Samples and Its Effect Evaluation

郑皓宇 郑文学 谷奕诺 安一鸣 张嘉琪 张雨涵 张姗姗 孟晓婷 王放
国际老年医学杂志2024,Vol.45Issue(2) :135-141.DOI:10.3969/j.issn.1674-7593.2024.02.002

快速定性检测人呼吸道样本中冠状病毒基因芯片的制备方法及其效果评价

Preparation Method of Gene Chip for Rapid Qualitative Detection of Coronavirus in Human Respiratory Samples and Its Effect Evaluation

郑皓宇 1郑文学 1谷奕诺 1安一鸣 1张嘉琪 1张雨涵 1张姗姗 1孟晓婷 2王放1
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作者信息

  • 1. 吉林大学基础医学院病原生物学教研室,长春 130021
  • 2. 吉林大学基础医学院组织学与胚胎学教研室,长春 130021
  • 折叠

摘要

目的 探讨冠状病毒基因芯片的研制方法,建立一种快速定性检测人咽拭子、痰液、肺泡灌洗液等呼吸道感染样本中的7种冠状病毒和2种流感病毒核酸的方法.方法 根据互联网公布的10种呼吸道感染病毒的基因序列,设计了相应的引物和探针序列,经琼脂糖凝胶电泳筛选验证后,将探针点样于醛基修饰基片,完成了检测芯片制作.结果 将聚合酶链式反应扩增产物用10倍梯度稀释法稀释后进行基因芯片检测,结果表明可形成杂交斑点的最低检测浓度为103copies/mL.结论 研制的冠状病毒基因芯片具有良好的特异度和灵敏度,可同时对SARS-CoV-2及其他呼吸道病毒同时进行高通量快速检测.

Abstract

Objective To explore the development method of coronavirus gene chip,and to establish a method for rapid qualitative detection of nucleic acids of 7 coronaviruses and 2 influenza viruses in respiratory infection samples such as human throat swabs,sputum,and alveolar lavage fluid.Methods According to the gene sequences of 10 kinds of respiratory infection viruses published on the Internet,the corresponding primers and probe sequences were designed.After screening and verification by agarose gel electrophoresis,the probes were spotted on the aldehyde base modified substrate,completed the detection chip fabrication.Re-sults The PCR amplification product was diluted by 10-fold serial dilution method and then detected by gene chip.The results showed that the lowest detection concentration that could form hybridization spots was 103copies/mL.Conclusion The developed coronavirus gene chip has good specificity and sensitivity,and can be used for simultaneous high-throughput and rapid detection of SARS-CoV-2 and other respiratory viruses.

关键词

冠状病毒/流感病毒/基因芯片

Key words

Coronavirus/Influenza virus/Gene chip

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基金项目

吉林省教育厅科学技术研究项目(JJKH20211158KJ)

吉林省科技发展项目自主课题(20200601011JC)

吉林省发改委自主创新能力课题(2022C036)

出版年

2024
国际老年医学杂志
吉林大学

国际老年医学杂志

CSTPCD
影响因子:0.435
ISSN:1674-7593
参考文献量25
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