[1]孙 璇,昝明辉,王慧慧,等.MIRA-CRISPR/Cas13a技术快速检测B族链球菌方法的建立及评价[J].现代检验医学杂志,2026,41(02):1-4+9.[doi:10.3969/j.issn.1671-7414.2026.02.001]
　SUN Xuan,ZAN Minghui,WANG Huihui,et al.Establishment and Evaluation of a Rapid Detection Method for Group B Streptococcus using MIRA-CRISPR/Cas13a Technology[J].Journal of Modern Laboratory Medicine,2026,41(02):1-4+9.[doi:10.3969/j.issn.1671-7414.2026.02.001]
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MIRA-CRISPR/Cas13a技术快速检测B族链球菌方法的建立及评价()

《现代检验医学杂志》[ISSN:/CN:]

卷:
第41卷
期数:
2026年02期
页码:
1-4+9
栏目:
出版日期:
2026-03-15

文章信息/Info

Title:
Establishment and Evaluation of a Rapid Detection Method for Group B Streptococcus using MIRA-CRISPR/Cas13a Technology
文章编号:
1671-7414（2026）02-001-05
作者:
孙 璇1，昝明辉2，王慧慧3,4，郑伟港3,4
1.北京市海淀区妇幼保健院，北京 100080;2.中国科学院苏州生物医学工程技术研究所，江苏苏州 215004;3.郑州中科生物医学工程技术研究院，郑州 450001;4.河南省中科科技成果转移转化中心，郑州 450003
Author(s):
SUN Xuan1，ZAN Minghui2，WANG Huihui3,4，ZHENG Weigang3,4
1.Beijing Haidian District Maternal and Child Health Care Hospital, Beijing 100080, China;2.Suzhou Institute of Biomedical Engineering and Technology, Chinese Academy of Sciences, Jiangsu Suzhou 215004, China;3.Zhengzhou Institute of Biomedical Engineering and Technolo-gy, Zhengzhou 450001, China;4.Henan Provincial Science and Technology Achievements Transfer and Transformation Center, Zhengzhou 450003, China
关键词:
B族链球菌; 多酶恒温快速扩增技术; 成簇规律间隔短回文重复序列
分类号:
R378.12；R446.19
DOI:
10.3969/j.issn.1671-7414.2026.02.001
文献标志码:
A
摘要:
目的?基于一管法多酶恒温快速扩增(MIRA)和成簇规律间隔短回文重复及其序列相关蛋白13a(CRISPR/Cas13a)技术开发一种兼具快速、准确和简单的B族链球菌(GBS)的检测方法。方法将MIRA的高效扩增与CRISPR/Cas13a的高特异性识别能力相结合,并将两个过程结合在同一反应管内进行,建立MIRA等温扩增和CRISPR/Cas13a切割的一管法GBS检测体系,并评价其灵敏度、特异度与实时荧光定量PCR(RT-qPCR)结果的一致性。结果成功建立一管法MI-RA-CRISPR/Cas13a检测体系。该方法可在40min内完成检测,灵敏度达到102copies/μl,与其他常见病原菌以及单、双碱基突变的非靶标序列无明显交叉反应,具有高度特异性。该方法与RT-qPCR方法的一致性为97.62%,Kappa值为0.949。结论建立的一管法MIRA-CRISPR/Cas13a检测体系具有快速、灵敏、特异、操作简便等优点,为GBS的快速筛查和诊断提供新的技术手段。
Abstract:
Objective A rapid, accurate and simple detection method for group B Streptococcus (GBS) was developed based on the one-tube multi-enzyme isothermal rapid amplification (MIRA) and clustered regularly interspaced short palindromic repeats (CRISPR) and their associated protein 13a (MIRA-CRISPR/Cas13a) technology. Methods Integrating the efficient amplifica-tion of MIRA with the highly specific recognition ability of CRISPR/Cas13a into a single reaction tube to establish a one-tube MIRA-CRISPR/Cas13a cleavage-based GBS detection system. Its sensitivity, specificity and consistency with RT-qPCR results were evaluated. Results A one-tube MIRA-CRISPR/Cas13a detection system was successfully established. The method could complete the detection within 40 minutes with a sensitivity of 102 copies/μl. It exhibited high specificity with no significant cross-reactivity to other common pathogens and non-targets sequences with single and double base mutations. The consistency rate between this method and RT-qPCR method was 97.62%, with a Kappa value of 0.949. Conclusions The one-tube MI-RA-CRISPR/Cas13a detection system established in this study offers advantages of rapidity, sensitivity, specificity and ease of operation, which provides a novel technical means for rapid screening and diagnosis of GBS.

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备注/Memo

备注/Memo:
基金项目：国家自然科学基金（82172077）；中科院河南中心资助基金（2024111）；苏州市基础研究试点项目（SSD2023015）。
作者简介：孙璇（1983-），女，学士，主管检验师，研究方向：免疫及生化检验，E-mail：xuaner0731@126.com。
通讯作者： 昝明辉（1988-），男，博士研究生，副研究员，研究方向：纳米生物医学工程技术，E-mail：minghui_zan@163.com。
郑伟港（1996-），男，硕士研究生，助理工程师，研究方向：分子学诊断，E-mail：zwg34@163.com。
更新日期/Last Update: 2026-03-15