iScience (Apr 2024)
Cultivation and sequencing-free protocol for Serratia marcescens detection and typing
- Alessandro Alvaro,
- Aurora Piazza,
- Stella Papaleo,
- Matteo Perini,
- Ajay Ratan Pasala,
- Simona Panelli,
- Tiago Nardi,
- Riccardo Nodari,
- Lodovico Sterzi,
- Cristina Pagani,
- Cristina Merla,
- Daniele Castelli,
- Emanuela Olivieri,
- Silvia Bracco,
- Maria Laura Ferrando,
- Francesca Saluzzo,
- Sara Giordana Rimoldi,
- Marta Corbella,
- Annalisa Cavallero,
- Paola Prati,
- Claudio Farina,
- Daniela Maria Cirillo,
- Gianvincenzo Zuccotti,
- Francesco Comandatore
Affiliations
- Alessandro Alvaro
- Department of Biomedical and Clinical Sciences, Pediatric Clinical Research Center “Romeo and Enrica Invernizzi”, University of Milan, 20157 Milan, Italy; Department of Biosciences and Pediatric Clinical Research Center ''Romeo Ed Enrica Invernizzi'', University of Milan, 20133 Milan, Italy
- Aurora Piazza
- Unit of Microbiology and Clinical Microbiology, Department of Clinical-Surgical, Diagnostic and Pediatric Sciences, University of Pavia, Pavia 27100, Italy
- Stella Papaleo
- Department of Biomedical and Clinical Sciences, Pediatric Clinical Research Center “Romeo and Enrica Invernizzi”, University of Milan, 20157 Milan, Italy
- Matteo Perini
- Department of Biomedical and Clinical Sciences, Pediatric Clinical Research Center “Romeo and Enrica Invernizzi”, University of Milan, 20157 Milan, Italy; Department of Environmental Health Sciences, Mailman School of Public Health, Columbia University, New York, NY 10032, USA
- Ajay Ratan Pasala
- Department of Biomedical and Clinical Sciences, Pediatric Clinical Research Center “Romeo and Enrica Invernizzi”, University of Milan, 20157 Milan, Italy; Biochemistry, Microbiology and Immunology Department, Faculty of Medicine, University of Ottawa, Ottawa, ON K1H 8M5, Canada; Centre for Innovation, Canadian Blood Services, Ottawa, ON K1G 4J5, Canada
- Simona Panelli
- Department of Biomedical and Clinical Sciences, Pediatric Clinical Research Center “Romeo and Enrica Invernizzi”, University of Milan, 20157 Milan, Italy
- Tiago Nardi
- Department of Biomedical and Clinical Sciences, Pediatric Clinical Research Center “Romeo and Enrica Invernizzi”, University of Milan, 20157 Milan, Italy; Department of Biology and Biotechnology, University of Pavia, 27100 Pavia, Italy
- Riccardo Nodari
- Department of Biomedical and Clinical Sciences, Pediatric Clinical Research Center “Romeo and Enrica Invernizzi”, University of Milan, 20157 Milan, Italy
- Lodovico Sterzi
- Department of Biomedical and Clinical Sciences, Pediatric Clinical Research Center “Romeo and Enrica Invernizzi”, University of Milan, 20157 Milan, Italy
- Cristina Pagani
- Laboratorio di Microbiologia Clinica, Virologia e Diagnostica delle Bioemergenze, ASST Fatebenefratelli Sacco, 20157 Milan, Italy
- Cristina Merla
- Department of Microbiology & Virology, Fondazione IRCCS Policlinico San Matteo, Viale Camillo Golgi 19, 27100 Pavia, Italy
- Daniele Castelli
- Microbiology Unit, Fondazione IRCCS San Gerardo, 20900 Monza, Italy
- Emanuela Olivieri
- Istituto Zooprofilattico Sperimentale della Lombardia e dell'Emilia Romagna (IZSLER), 27100 Pavia, Italy
- Silvia Bracco
- Laboratory of Microbiology and Virology, Azienda Socio-Sanitaria Territoriale (ASST) Papa Giovanni XXIII, 24127 Bergamo, Italy
- Maria Laura Ferrando
- Emerging Bacterial Pathogens Unit, Division of Immunology, Transplantation and Infectious Diseases, IRCCS San Raffaele Scientific Institute, 20132 Milan, Italy
- Francesca Saluzzo
- Emerging Bacterial Pathogens Unit, Division of Immunology, Transplantation and Infectious Diseases, IRCCS San Raffaele Scientific Institute, 20132 Milan, Italy
- Sara Giordana Rimoldi
- Laboratorio di Microbiologia Clinica, Virologia e Diagnostica delle Bioemergenze, ASST Fatebenefratelli Sacco, 20157 Milan, Italy
- Marta Corbella
- Department of Microbiology & Virology, Fondazione IRCCS Policlinico San Matteo, Viale Camillo Golgi 19, 27100 Pavia, Italy
- Annalisa Cavallero
- Microbiology Unit, Fondazione IRCCS San Gerardo, 20900 Monza, Italy
- Paola Prati
- Istituto Zooprofilattico Sperimentale della Lombardia e dell'Emilia Romagna (IZSLER), 27100 Pavia, Italy
- Claudio Farina
- Laboratory of Microbiology and Virology, Azienda Socio-Sanitaria Territoriale (ASST) Papa Giovanni XXIII, 24127 Bergamo, Italy
- Daniela Maria Cirillo
- Emerging Bacterial Pathogens Unit, Division of Immunology, Transplantation and Infectious Diseases, IRCCS San Raffaele Scientific Institute, 20132 Milan, Italy
- Gianvincenzo Zuccotti
- Department of Biomedical and Clinical Sciences, Pediatric Clinical Research Center “Romeo and Enrica Invernizzi”, University of Milan, 20157 Milan, Italy; Department of Paediatrics, Children’s Hospital ''V. Buzzi'', 20154 Milano, Italy
- Francesco Comandatore
- Department of Biomedical and Clinical Sciences, Pediatric Clinical Research Center “Romeo and Enrica Invernizzi”, University of Milan, 20157 Milan, Italy; Corresponding author
- Journal volume & issue
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Vol. 27,
no. 4
p. 109402
Abstract
Summary: Serratia marcescens is an opportunistic pathogen that survives in inhospitable environments causing large outbreaks, particularly in neonatal intensive care units (NICUs). Genomic studies revealed that most S. marcescens nosocomial infections are caused by a specific clone (here “Infectious clone”). Whole genome sequencing (WGS) is the only portable method able to identify this clone, but it requires days to obtain results. We present a cultivation-free hypervariable-locus melting typing (HLMT) protocol for the fast detection and typing of S. marcescens, with 100% detection capability on mixed samples and a limit of detection that can reach the 10 genome copies. The protocol was able to identify the S. marcescens infectious clone with 97% specificity and 96% sensitivity when compared to WGS, yielding typing results portable among laboratories. The protocol is a cost and time saving method for S. marcescens detection and typing for large environmental/clinical surveillance screenings, also in low-middle income countries.
