Acute gastroenteritis (AGE) still remains one of the most common diseases in humans worldwide, being responsible for about half a million deaths in children aged < 5 years, especially in developing countries (Troeger et al., 2017). In industrialized nations, although less fatal, AGE imposes a significant burden on healthcare systems due to medical expenses, productivity loss, and social costs (Peery et al., 2022). AGE can be triggered by a wide range of infectious agents, and clinical presentation alone is often insufficient to differentiate between viral and bacterial etiologies (Bányai et al., 2018, Hassan et al., 2019). Children under 5 years are particularly susceptible to viral AGE and are more often affected by severe symptoms, including vomiting and /or watery diarrhea, leading to life-threatening dehydration, and requiring hospitalization.

Accurate aetiological diagnosis is important for proper clinical management and to implement preventive measures to control virus spread. Moreover, identifying the specific viral pathogen is crucial to avoid unnecessary antimicrobial use and to inform future preventive strategies, including vaccination programs (Danino et al., 2023).

Norovirus (NoV) and group A rotavirus (RVA) account for almost half of viral AGE cases in children, followed by adenovirus F40/41 (AdV), human astrovirus (HAstV) and sapovirus (SV) (Bányai et al., 2018, Hassan et al., 2019, Oude Munnink and van der Hoek, 2016, Zhang et al., 2011).

Recently, several molecular syndromic assays have been introduced, offering simultaneous detection of the most relevant viral AGE pathogens. These assays, based on different chemistries, have replaced traditional diagnostic methods, streamlining laboratory workflows, enhancing analytical sensitivity, and improving diagnostic accuracy (Danino et al., 2023, De Grazia et al., 2019, Buss et al., 2015, Khamrin et al., 2011). However, their performance must be periodically reassessed to ensure continued effectiveness in detecting pathogens amid evolving epidemiological scenarios. This study aimed to value the diagnostic performance of two newly proposed version of multiplex PCR assays, the GI Viral PLUS ELITe MGB and GI Norovirus PLUS ELITe MGB® (ELITechGroup Molecular Diagnostics, Puteaux, France) using a broad panel of clinically and epidemiologically relevant viral genotypes.