验证一种环介导等温扩增检测法，用于快速诊断鼻咽样本中的百日咳感染。
Validation of a loop-mediated isothermal amplification assay for rapid diagnosis of pertussis infection in nasopharyngeal samples.
机构信息
a Department of Molecular Microbiology , Hospital Sant Joan de Déu, University of Barcelona , Barcelona , Spain.
出版信息
Expert Rev Mol Diagn. 2016;16(1):125-30. doi: 10.1586/14737159.2016.1112741. Epub 2015 Nov 13.
OBJECTIVE
To develop and validate a novel loop-mediated amplification (LAMP) assay for rapid diagnosis (<1 hour) of whooping cough in nasopharyngeal samples versus the gold standard: real-time PCR.
METHODS
The study included all nasopharyngeal samples (n = 213) collected from children with clinical suspicion of pertussis admitted to Children's University Hospital Sant Joan de Déu (Barcelona, Spain) during July-December 2014. Fresh samples were routinely analyzed by real-time PCR and stored for retrospective LAMP analysis, following an easy 30 minute DNA extraction step by Chelex-100.
RESULTS
Performance results of the LAMP assay were: linearity, 10(5)-10(1) CFU/ml; Limit of Detection, 2 CFU/ml; precision (mean CV), 7.38%; diagnostic sensitivity, 96.55%; diagnostic specificity, 99.46%; time to detection, 12-30 minutes.
CONCLUSION
The new test was shown to be 2.5-fold faster than real-time PCR while maintaining similar levels of analytical and clinical performance. Therefore it could become a useful diagnostic tool for molecular point-of-care testing.
目的
开发并验证一种新型环介导扩增（LAMP）检测法，用于快速诊断（<1 小时）鼻咽样本中的百日咳，与金标准实时聚合酶链反应（PCR）相比。
方法
该研究纳入了 2014 年 7 月至 12 月期间因临床疑似百日咳而入住西班牙巴塞罗那桑特·乔迪儿童大学医院的所有儿童鼻咽样本（n=213）。新鲜样本通常通过实时 PCR 进行常规分析，并在通过 Chelex-100 进行简单的 30 分钟 DNA 提取步骤后，用于回顾性 LAMP 分析。
结果
LAMP 检测法的性能结果如下：线性范围为 10(5)-10(1)CFU/ml；检测限为 2CFU/ml；精密度（平均 CV）为 7.38%；诊断灵敏度为 96.55%；诊断特异性为 99.46%；检测时间为 12-30 分钟。
结论
新检测法比实时 PCR 快 2.5 倍，同时保持类似的分析和临床性能。因此，它可能成为分子即时检测的有用诊断工具。
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