可视化LAMP检测PJI金黄色葡萄球菌的感染

Visual detection of staphylococcus aureus from periprosthetic joint infection by loop-mediated isothermal amplification assay

目的为建立一种可早期明确假体周围感染(PJI)菌种的快速、可视化检测方法。方法根据金黄色葡萄球菌基因中的保守序列设计特异性引物,以甲酚红为指示剂,评估该方法的灵敏度和特异度,并进行临床样本验证。结果可视化环介导等温扩增(LAMP)检测可在60 min(样本处理3~5 min,核酸提取15 min,可视化LAMP检测40 min)内完成对样本的检测,其最低检测限为10 copies/μl,且具有良好的特异性,在对临床关节液样本进行符合率验证时,其阳性符合率和阴性符合率均为100%。结论可视化LAMP检测在PJI感染检测中显示出良好的应用潜力。

Objective In order to establish a rapid and visual detection method for early identification of periprosthetic infection bacteria.Methods Specific primers were designed according to the conserved sequences in the genes of staphylococcus aureus,and cresyl red was used as an indicator to evaluate the sensitivity and specificity of the method,and to conduct clinical sample validation.Results This study found that the visual LAMP(loop-mediated isothermal amplification)detection can complete the sample detection within 60 minutes(3~5 minutes for sample processing,15 minutes for nucleic acid extraction,and 40 minutes for visual LAMP detection).The minimum detection limit of sensitivity is 10 copies/μl,and this detection has good specificity.When verifying the coincidence rate of clinical synovial fluid samples,the positive and negative coincidence rates are both 100%.Conclusion This study shows that the visual LAMP detection has good application potential in PJI infection detection.