Adapting an in-house viral gastroenteritis multiplex PCR for use on the Hologic Panther Fusion® system

Viral gastroenteritis is a major global health concern, causing billions of infections annually. Multiplex real-time PCR assays for gastro-intestinal viruses (GIV) ensure timely diagnosis, enabling clinical management, improved antimicrobial stewardship and infection prevention actions and supports surveillance. To improve GIV diagnostics used in our tertiary hospital, we adapted and validated our current manual in-house in vitro diagnostic (IH-IVD) PCR panel for GIV on the Hologic Panther Fusion® system (a high-throughput, fully automated sample-to-result IVD system with Open Access™ functionality).

We compared our current manual IH-IVD method on Roche Flow with the automated Hologic Panther Fusion® system using identical primers and probes targeting rota-, noro- (genotype GI and GII), adeno- and adeno- F (type 40-41), astro-, sapo-, entero- and parechovirus. For clinical validation, 301 fecal samples collected between 2015 and 2024 were analyzed with the Hologic Panther Fusion® system and compared with prior IH-IVD assay results. Analytical sensitivity was assessed using serial dilutions of viral stocks and an external quality assessment (EQA) panel by Quality Control for Molecular Diagnostics (QCMD).

Clinical sensitivity and specificity ranged from 91.3% to 100% and 99.3-100%, respectively. Analytical sensitivity was slightly lower for Panther Fusion® (10-fold), except for adeno-F- and norovirus genotype GII which showed higher sensitivity (5-fold). EQA panels were concordant with both systems and the expected QCMD outcomes.

The Panther Fusion® is a fully automated system with minimal hands-on time and rapid sample-to-result turnaround, which offers a reliable outcome despite slightly lower analytical sensitivity compared to IH-IVD.