Enteroviruses (EVs) represent an important public health problem, as they are responsible for respiratory and systemic infections that can manifest themselves with even serious clinical symptoms, especially in pediatric and vulnerable patients. Their high genetic variability, due to the low fidelity of RNA replication and frequent recombination events, makes the implementation of molecular surveillance strategies aimed at the identification of new viral strains. The aim of the present study is the molecular characterization of enterovirus strains isolated from respiratory clinical specimens of patients with confirmed infection by routine diagnostic tests. The analysis was conducted through automated viral RNA extraction, cDNA synthesis, amplification by PCR, and Sanger sequencing, with extension to Next Generation Sequencing (NGS) technologies. The sequences obtained were subjected to bioinformatic analysis for genomic characterization of the identified strains. Finally, molecular data were integrated with descriptive statistical analysis and with demographic and clinical information of patients, in order to evaluate any associations useful for epidemiological surveillance and understanding of enterovirus circulation.
Gli enterovirus (EV) rappresentano un importante problema di salute pubblica, poiché sono responsabili di infezioni respiratorie e sistemiche che possono manifestarsi anche con sintomi clinici gravi, specialmente nei pazienti pediatrici e vulnerabili. La loro elevata variabilità genetica, dovuta alla bassa fedeltà della replicazione dell'RNA e ai frequenti eventi di ricombinazione, rende l'implementazione di strategie di sorveglianza molecolare volte all'identificazione di nuovi ceppi virali. L'obiettivo del presente studio è la caratterizzazione molecolare di ceppi di enterovirus isolati da campioni clinici respiratori di pazienti con infezione confermata tramite test diagnostici di routine. L'analisi è stata condotta tramite estrazione automatica di RNA virale, sintesi di cDNA, amplificazione tramite PCR e sequenziamento Sanger, con estensione alle tecnologie di Next Generation Sequencing (NGS). Le sequenze ottenute sono state sottoposte ad analisi bioinformatiche per la caratterizzazione genomica dei ceppi identificati. Infine, i dati molecolari sono stati integrati con analisi statistiche descrittive e con informazioni demografiche e cliniche dei pazienti, al fine di valutare eventuali associazioni utili alla sorveglianza epidemiologica e alla comprensione della circolazione dell'enterovirus.
Analisi e caratterizzazione genomica di enterovirus in campioni clinici respiratori
RUSSO, MARTINA
2025/2026
Abstract
Enteroviruses (EVs) represent an important public health problem, as they are responsible for respiratory and systemic infections that can manifest themselves with even serious clinical symptoms, especially in pediatric and vulnerable patients. Their high genetic variability, due to the low fidelity of RNA replication and frequent recombination events, makes the implementation of molecular surveillance strategies aimed at the identification of new viral strains. The aim of the present study is the molecular characterization of enterovirus strains isolated from respiratory clinical specimens of patients with confirmed infection by routine diagnostic tests. The analysis was conducted through automated viral RNA extraction, cDNA synthesis, amplification by PCR, and Sanger sequencing, with extension to Next Generation Sequencing (NGS) technologies. The sequences obtained were subjected to bioinformatic analysis for genomic characterization of the identified strains. Finally, molecular data were integrated with descriptive statistical analysis and with demographic and clinical information of patients, in order to evaluate any associations useful for epidemiological surveillance and understanding of enterovirus circulation.| File | Dimensione | Formato | |
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https://hdl.handle.net/20.500.12608/117064