| 蒋婷清,杨涛,唐红花,王萍,何军,邓仲良.ERA实时荧光法快速检测鹦鹉热衣原体方法的建立.[J].中南医学科学杂志.,2023,(4):502-506. |
| ERA实时荧光法快速检测鹦鹉热衣原体方法的建立 |
| Establishment and application of ERA real-time fluorescence method for rapid detection of Chlamydia psittaci |
| 投稿时间：2023-02-22 修订日期：2023-06-30 |
| DOI：10.15972/j.cnki.43-1509/r.2023.04.007 |
| 中文关键词: 酶促重组等温扩增 鹦鹉热衣原体 实时荧光检测 [ |
| 英文关键词:enzymatic recombinase amplification Chlamydia psittaci real-time fluorescence detection |
| 基金项目:湖南省教育厅重点项目(21A0261),湖南省自然科学基金项目(2022JJ30501) |
| 作者 | 单位 | E-mail | | 蒋婷清 | 南华大学衡阳医学院 公共卫生学院,湖南衡阳 421001 | e-mail为517945150@qq.com,e-mail为dzl021015@163.com,e-mail为junhe2008@163.com | | 杨涛 | 南华大学衡阳医学院 公共卫生学院,湖南衡阳 421001 | | | 唐红花 | 南华大学衡阳医学院 附属第一医院,湖南衡阳 421001 | | | 王萍 | 常德市武陵区疾病预防控制中心, 湖南常德415000 | | | 何军 | 南华大学衡阳医学院附属南华医院,湖南衡阳421002 | e-mail为517945150@qq.com,e-mail为dzl021015@163.com,e-mail为junhe2008@163.com | | 邓仲良 | 南华大学衡阳医学院 公共卫生学院,湖南衡阳 421001 | e-mail为517945150@qq.com,e-mail为dzl021015@163.com,e-mail为junhe2008@163.com |
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| 中文摘要: |
| 目的建立一种快速检测鹦鹉热衣原体(Cps)的酶促重组等温扩增(ERA)实时荧光法。 方法根据鹦鹉热衣原体MOMP基因序列设计特异性引物和探针,建立ERA实时荧光法反应体系。优化体系中的引物和反应温度,再评价ERA实时荧光法的敏感性和特异性,以及对39例鸟类肺组织样本进行验证。 结果ERA实时荧光法可在40 ℃恒温条件下15 min内特异性扩增鹦鹉热衣原体,其检出限为10 copies/μL,特异性好。ERA实时荧光法检出39例鸟类肺组织样本中阳性样本30例,阴性样本9例,其灵敏度为96.77%,特异度为100.00%,阴性预测值为88.89%,阳性预测值为100.00%。 结论成功建立用于鹦鹉热衣原体检测的ERA实时荧光法,其操作简单,且灵敏度和特异度好。 |
| 英文摘要: |
| AimTo establish an enzymatic recombinant isothermal amplification (ERA) real-time fluorescence assay for rapid detection of Chlamydia psittaci (Cps). MethodsThe ERA real-time fluorometric reaction system was established by designing specific primers and probes based on the Cps MOMP gene sequence. The ERA real-time fluorescence assay was optimized by adjusting the primers and reaction temperature, and its sensitivity and specificity were evaluated. Subsequently, 39 bird lung tissue samples were used for test. ResultsThe ERA real-time fluorescence method can specifically amplify Cps within 15 min at 40 ℃, with a detection limit of 10 copies/μL and strong specificity. The ERA real-time fluorescence method detected 30 positive samples and 9 negative samples in 39 bird lung tissue samples, with a sensitivity of 96.77%, specificity of 100.00%, negative predictive value of 88.89%, and positive predictive value of 100.00%. ConclusionThe ERA real-time fluorescent method has been established to identify Cps, and it is simple to operate, with excellent sensitivity and specificity. |
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