Raghavan Ramya, Wang Shouao, Dendukuri Nandini, Kar Sitanshu S, Mahadevan Subramanian, Jagadisan Barath, Mandal Jharna
Department of Microbiology, Jawaharlal Institute of Postgraduate Medical Education and Research, Puducherry, India.
Centre for Outcomes Research, McGill University Health Centre - Research Institute, 5252 Boulevard de Maisonneuve W, Montreal PQ H4A 3S5, Canada.
Access Microbiol. 2020 Sep 14;2(11):acmi000169. doi: 10.1099/acmi.0.000169. eCollection 2020.
To assess the diagnostic accuracy of loop-mediated isothermal amplification (LAMP) for the detection of from stool samples from children.
Consecutive stool samples from children aged <13 years old who presented with acute watery diarrhoea or dysentery to the Department of Paediatrics were collected and processed in the Department of Microbiology. All the stool samples were subjected to culture, conventional PCR and LAMP. Genomic sequencing was performed for samples that were positive by LAMP but negative by both culture and conventional PCR. The LAMP results were compared to those from culture and to a composite reference standard based on culture and conventional PCR.
Amongst the 374 stool samples tested, 291 samples were positive by LAMP and 213 were positive by the composite reference standard. The sensitivity of LAMP was 100 % (98.3-100 %) and its specificity was 51.6 % (43.6-59.5 %) with a disease prevalence of 57 %. The sensitivity and specificity of LAMP improved to 99.3 % (94.2-100) and 98.2 % (94.5-99.9), respectively, using latent class analysis, while assuming that genomic sequencing has perfect specificity.
The authors have standardized the LAMP procedure for direct application to clinical stool samples. LAMP is a sensitive and specific method for the diagnosis of from stool samples of children as compared to both culture and conventional PCR.
评估环介导等温扩增技术(LAMP)检测儿童粪便样本中[病原体名称未给出]的诊断准确性。
收集儿科就诊的13岁以下出现急性水样腹泻或痢疾的儿童的连续粪便样本,并在微生物科进行处理。所有粪便样本均进行培养、传统PCR和LAMP检测。对LAMP检测呈阳性但培养和传统PCR检测均呈阴性的样本进行基因组测序。将LAMP结果与培养结果以及基于培养和传统PCR的综合参考标准进行比较。
在检测的374份粪便样本中,291份样本LAMP检测呈阳性,213份样本综合参考标准检测呈阳性。疾病患病率为57%时,LAMP的灵敏度为100%(98.3 - 100%),特异性为51.6%(43.6 - 59.5%)。在假设基因组测序具有完美特异性的情况下,使用潜在类别分析,LAMP的灵敏度和特异性分别提高到99.3%(94.2 - 100)和98.2%(94.5 - 99.9)。
作者已将LAMP程序标准化,可直接应用于临床粪便样本。与培养和传统PCR相比,LAMP是一种检测儿童粪便样本中[病原体名称未给出]的灵敏且特异方法。