| 应用CRISPR/Cas13a快速鉴定布鲁氏菌 |
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| 引用本文： | 黄明耀,梁文立,吴婉婷,刘足,谢淑媚,邓颖颖,傅俊方,姜长宏,龙军,江凌晓. 应用CRISPR/Cas13a快速鉴定布鲁氏菌[J]. 中国人兽共患病杂志, 2021, 37(5): 426-429. DOI: 10.3969/j.issn.1002-2694.2021.00.062 |
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| 作者姓名： | 黄明耀 梁文立 吴婉婷 刘足 谢淑媚 邓颖颖 傅俊方 姜长宏 龙军 江凌晓 |
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| 作者单位： | 1. 南方医科大学珠江医院检验医学部, 广州 510282;2. 微远基因科技有限公司, 广州 510000;3. 微远医学研究中心有限公司, 深圳 518000 |
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| 基金项目： | 国家重点研发计划项目(No.2017YFC1200801) |
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| 摘 要： | 目的 建立一种基于CRISPR/Cas13a的布鲁氏菌鉴定方法.方法 以布鲁氏菌BCSP31基因的保守区为靶标区域设计特异的引物和crRNA,建立鉴定布鲁氏菌的CRISPR/Cas13a方法.通过对纯菌株以及临床需氧血培养阳性菌液检测评价该方法的特异性和敏感性;通过梯度稀释法评估其检测下限.结果 建立了基于CRISPR...
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| 关 键 词： | 布鲁氏菌 CRISPR/Cas13a 快速诊断 |
| 收稿时间： | 2020-11-27 |
Rapid identification of Brucella with CRISPR/Cas13a |
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| HUANG Ming-yao,LIANG Wen-li,WU Wan-ting,LIU Zu,XIE Shu-mei,DENG Ying-ying,FU Jun-fang,JIANG Chang-hong,LONG Jun,JIANG Ling-xiao. Rapid identification of Brucella with CRISPR/Cas13a[J]. Chinese Journal of Zoonoses, 2021, 37(5): 426-429. DOI: 10.3969/j.issn.1002-2694.2021.00.062 |
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| Authors: | HUANG Ming-yao LIANG Wen-li WU Wan-ting LIU Zu XIE Shu-mei DENG Ying-ying FU Jun-fang JIANG Chang-hong LONG Jun JIANG Ling-xiao |
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| Affiliation: | 1. Division of Laboratory Medicine, Zhujiang Hospital, Southern Medical University, Guangzhou 510282, China;2. Vision Medicals, Co., Ltd, Guangzhou 510000, China;3. Vision Medicals Center for Medical Research, Shenzhen 518000, China |
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| Abstract: | This study aimed to establish a method based on CRISPR/Cas13a to identify Brucella. We designed specific primers for RPA amplification targeting the conserved region of the Brucella BCSP31 gene to develop a CRISPR/Cas13a method to identify Brucella. The specificity and sensitivity of the method were evaluated by testing pure strains and positive clinical aerobic blood cultures; the lower limit of this method was evaluated with the gradient dilution method. A rapid identification method for Brucella through CRISPR/Cas13a was successfully established, and its detection limit reached 10 fg/μL. On the basis of detection of 64 Brucella strains, 56 non-Brucella strains, human DNA, and 57 cases of clinical aerobic blood culture positive bacteria, the sensitivity and specificity of this method reached 100%. In conclusion, a Brucella identification method based on CRISPR/Cas13a was established and can be applied for rapid identification of Brucella after a positive clinical aerobic blood culture. |
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| Keywords: | Brucella CRISPR/Cas13a rapid diagnosis |
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