| Autor: |
Caruso G; U.O.C. of Microbiology and Virology, ARNAS 'Civico Di Cristina and Benfratelli', 90127 Palermo, Italy., Coniglio MA; Legionella Reference Laboratory, Department of Medical, Surgical Sciences and Advanced Technologies 'G.F. Ingrassia', University of Catania, Via Santa Sofia 87, 95123 Catania, Italy.; Hygiene Complex Operative Unit, A.O.U. Policlinico-Vittorio Emanuele, Via S. Sofia 87, 95123 Catania, Italy., Laganà P; Legionella Reference Laboratory, University of Messina, 98125 Messina, Italy.; Department of Biomedical and Dental Sciences and Morphofunctional Imaging, University of Messina, 98125 Messina, Italy., Fasciana T; Legionella Reference Laboratory, University of Palermo, 90127 Palermo, Italy.; Department of Health Promotion, Mother and Child Care, Internal Medicine and Medical Specialties, University of Palermo, 90127 Palermo, Italy., Arcoleo G; Enbiotech s.r.l., Via Aquileia 34, 90144 Palermo, Italy., Arrigo I; Legionella Reference Laboratory, University of Palermo, 90127 Palermo, Italy., Di Carlo P; Department of Health Promotion, Mother and Child Care, Internal Medicine and Medical Specialties, University of Palermo, 90127 Palermo, Italy., Palermo M; Sicilian Health Department, Public Health and Environmental Risks Service, 90127 Palermo, Italy., Giammanco A; Legionella Reference Laboratory, University of Palermo, 90127 Palermo, Italy.; Department of Health Promotion, Mother and Child Care, Internal Medicine and Medical Specialties, University of Palermo, 90127 Palermo, Italy. |
| Abstrakt: |
Legionella pneumophila is a freshwater opportunistic pathogen and the leading cause of severe pneumonia known as Legionnaires' disease. It can be found in all water systems and survives in biofilms, free-living amoebae, and a wide variety of facilities, such as air conditioning and showers in hospitals, hotels and spas. The reference cultural method allows for the isolation and identification in many days, and in addition, it does not detect viable but rather non-culturable bacteria, increasing the risk of infection. In this context, a new LAMP-based (loop-mediated isothermal amplification) kit was developed, allowing for the rapid, sensitive, and labor-saving detection of L. pneumophila . The kit, " Legionella pneumophila Glow", was validated according to ISO/TS 12869:2012, testing sensitivity, inclusivity and exclusivity, and kit robustness. Sensitivity showed that the " Legionella pneumophila Glow" kit can detect up to 28 plasmid copies/µL. Robustness tests showed consistent results, with both contamination levels and the matrices used giving reproducible results. Furthermore, real samples were evaluated to compare the performance of the two methods. The LAMP kit " Legionella pneumophila Glow" proved a useful option for the rapid, efficient, and labor-saving screening of different typologies of water samples, offering significant advantages over the traditional method, as it is characterized by a high sensitivity, ease of use for laboratory testing, and a large reduction in analysis time, making it an asset to official controls. |
