Fast immunosensing technique to detect Legionella pneumophila in different natural and anthropogenic environments: comparative and collaborative trials

BMC Microbiology

Guillermo Rodríguez⁴  Eva Baldrich¹  Roberto Fernández-Lafuente²  Inmaculada Solís³  Sonia Macián⁴  Begoña Bedrina⁴ 

[1] Institut de Microelectrònica (IMB-CNM, CSIC), Campus Univ. Autònoma de Barcelona, Bellaterra, Barcelona, 08193, Spain;Departamento de Biocatálisis, Instituto de Catálisis y Petroleoquímica, Consejo Superior de Investigaciones Científicas, Campus UAM-CSIC, Cantoblanco, Madrid, 28049, Spain;Iproma, S.L, Cno.de la Raya 46, Castellón, 12005, Spain;Biótica, Bioquímica Analítica, S.L, Science and Technology Park of Jaume I University, Campus Riu Sec - Espaitec 2, planta baja, Castellón de la Plana, E12071, Spain

关键词: Magnetic particles;    Immunosensing;    Detection;    Legionella;   

Others  :  1143866
DOI  :  10.1186/1471-2180-13-88

 received in 2012-11-15, accepted in 2013-04-16,  发布年份 2013

【 摘 要 】

Background

Legionellosis is an uncommon form of pneumonia. After a clinical encounter, the necessary antibiotic treatment is available if the diagnosis is made early in the illness. Before the clinical encounter, early detection of the main pathogen involved, Legionella pneumophila, in hazardous environments is important in preventing infectious levels of this bacterium. In this study a qualitative test based on combined magnetic immunocapture and enzyme-immunoassay for the fast detection of Legionella pneumophila in water samples was compared with the standard method, in both comparative and collaborative trials. The test was based on the use of anti-Legionella pneumophila antibodies immobilized on magnetic microspheres. The final protocol included concentration by filtration, resuspension and immunomagnetic capture. The whole assay took less than 1 hour to complete.

Results

A comparative trial was performed against the standard culture method (ISO 11731) on both artificially and naturally contaminated water samples, for two matrices: chlorinated tap water and cooling tower water. Performance characteristics of the test used as screening with culture confirmation resulted in sensitivity, specificity, false positive, false negative, and efficiency of 96.6%, 100%, 0%, 3.4%, and 97.8%, respectively. The detection limit at the level under which the false negative rate increases to 50% (LOD50) was 93 colony forming units (CFU) in the volume examined for both tested matrices. The collaborative trial included twelve laboratories. Water samples spiked with certified reference materials were tested. In this study the coincidence level between the two methods was 95.8%.

Conclusion

Results demonstrate the applicability of this immunosensing technique to the rapid, simple, and efficient detection of Legionella pneumophila in water samples. This test is not based on microbial growth, so it could be used as a rapid screening technique for the detection of L. pneumophila in waters, maintaining the performance of conventional culture for isolation of the pathogen and related studies.

【 授权许可】

   
2013 Bedrina et al.; licensee BioMed Central Ltd.

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