Unitat de Malalties Infeccioses, Fundació Institut d'Investigació Germans Trias i Pujol, Badalona E-08916, Spain; CIBER de Enfermedades Respiratorias, Madrid E-28029, Spain.
Unitat de Malalties Infeccioses, Fundació Institut d'Investigació Germans Trias i Pujol, Badalona E-08916, Spain.
Talanta. 2018 Nov 1;189:324-331. doi: 10.1016/j.talanta.2018.07.013. Epub 2018 Jul 7.
Waterborne pathogens are a global concern for public health worldwide. Despite continuing efforts to maintain water safety, water quality is still affected by deterioration and pollution. Legionella pneumophila colonizes man-made water systems and can infect humans causing Legionnaire's disease (LD), pneumonia. The prevention of LD is a public health issue and requires specific systems to control and detect these microorganisms. Culture plate is the only technique currently approved, but requires more than 10 days to obtain results. A rapid test that inform in hours about the presence of Legionella pneumophila in water samples will improve the control of this pathogen colonization. In order to control colonization by L. pneumophila we developed a membrane filter method to capture and immunodetect this microorganism in water samples. This membrane filter is used to retain the bacteria using a nitrocellulose disc inside a home-made cartridge. Subsequently we perform the immunodetection of the bacteria retained in the nitrocellulose (blocking, antibody incubation, washings and developing). On comparing our test with the gold-standard, the most important finding is the considerably reduction in time maintaining the same detection limit. This rapid test is easily automated for L. pneumophila detection allowing a comprehensive surveillance of L. pneumophila in water facilities and reducing the variability in the analyses due to the low need for manipulation. Moreover, corrective measures may be applied the same day of the analysis. This method considerably reduces the detection time compared with the conventional, gold-standard detection culture method that requires more than 10 days, being decisive to prevent outbreaks.
水传播病原体是全球公共卫生关注的问题。尽管在维护水安全方面不断努力,但水质仍受到恶化和污染的影响。嗜肺军团菌在人造水系统中定殖,并可感染人类引起军团病(LD),肺炎。LD 的预防是一个公共卫生问题,需要特定的系统来控制和检测这些微生物。培养板是目前唯一批准的技术,但需要 10 天以上才能获得结果。一种能够在数小时内告知水中是否存在嗜肺军团菌的快速检测方法将改善对这种病原体定殖的控制。为了控制嗜肺军团菌的定殖,我们开发了一种膜过滤器方法来捕获和免疫检测水样中的这种微生物。该膜过滤器用于使用家用试剂盒内的硝酸纤维素盘保留细菌。随后,我们对保留在硝酸纤维素上的细菌进行免疫检测(封闭、抗体孵育、洗涤和显色)。与金标准相比,我们的测试最重要的发现是在保持相同检测限的情况下,时间大大缩短。这种快速检测方法很容易实现嗜肺军团菌的自动化检测,从而可以全面监测水中设施中的嗜肺军团菌,并由于操作需求低而减少分析中的变异性。此外,还可以在分析当天采取纠正措施。与传统的、需要 10 天以上的金标准培养检测方法相比,该方法大大缩短了检测时间,对于预防爆发至关重要。