JCLA：通过重组酶-聚合酶扩增- sybr green I法肉眼检测结核分枝杆菌复合物

2019-06-03 不详网络

结核分枝杆菌(Mtb)的快速诊断是控制结核病传播的关键，也是全球健康问题。本研究利用等温重组酶聚合酶扩增(RPA)技术检测了Mtb、IS6110和IS1081的特异性靶点。此外，使用肉眼检测了SYBR Green I的RPA产物。研究人员利用RPA分别在IS6110和IS1081位点扩增了146份基因组Mtb DNA和24份基因组非结核分枝杆菌(NTM) DNA。完成扩增后，用琼脂糖凝胶电

结核分枝杆菌(Mtb)的快速诊断是控制结核病传播的关键，也是全球健康问题。本研究利用等温重组酶聚合酶扩增(RPA)技术检测了Mtb、IS6110和IS1081的特异性靶点。此外，使用肉眼检测了SYBR Green I的RPA产物。

研究人员利用RPA分别在IS6110和IS1081位点扩增了146份基因组Mtb DNA和24份基因组非结核分枝杆菌(NTM) DNA。完成扩增后，用琼脂糖凝胶电泳(RPA‐AGE)和SYBR Green I (RPA‐S)检测RPA扩增子。通过与传统PCR方法的比较，评价了RPA法的性能。

结果表明，RPA法在恒温条件下具有较好的鉴别Mtb和NTM的能力，且在很短的时间内就能鉴别出Mtb和NTM。与传统PCR方法相比，RPA‐年龄对IS6110和IS1081的敏感性和特异性均为100%。RPA‐S对两种靶点的特异性均为100%；但对IS6110和IS1081的敏感性分别为97.95%和99.32%。IS6110 RPA‐年龄和RPA‐S的检测限分别为0.05和0.5 ng，而IS1081 RPA‐年龄和RPA‐S的LODs分别为0.00005和0.05 ng。两种RPA检测均显示出令人满意的诊断特异性，与其他细菌无交叉反应。

研究表明，这种快速、灵敏、裸眼RPA检测方法可以集成到Mtb检测的定点护理诊断中，特别是在实验室仪器资源有限的偏远地区。

原始出处：

Nuntita Singpanomchai, Yukihiro Akeda ,Naked eye detection of the Mycobacterium tuberculosis complex by recombinase polymerase amplification—SYBR green I assays

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2019-08-22 yang0210

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2019-11-11 晓辰

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2019-06-30 lixiaol

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2019-06-05 huhuaidong387

#结核#

59 0
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