Enhanced enteric virus detection in sporadic gastroenteritis using a multi-target real-time PCR panel: a one-year...

Area of Science:

• Virology
• Molecular Biology
• Public Health

Background:

• Viral gastroenteritis is a major global health concern with significant mortality and morbidity.
• Accurate etiological diagnosis of viral gastroenteritis is challenging due to the non-culturable nature of most enteric viruses.
• Electron microscopy (EM) has limitations in detecting various enteric viruses.

Purpose of the Study:

• To develop and evaluate a real-time PCR (EVPrtPCR) panel for simultaneous detection of common enteric viruses in stool samples.
• To compare the diagnostic performance of EVPrtPCR with electron microscopy (EM) for identifying viral gastroenteritis etiologies.
• To determine the prevalence and demographic distribution of different enteric viruses in sporadic gastroenteritis cases.

Main Methods:

• Development of a multiplex real-time PCR assay (EVPrtPCR) for rotavirus, norovirus, sapovirus, astrovirus, and enteric adenovirus.
• Testing of 2,486 sporadic gastroenteritis stool samples using both EVPrtPCR and electron microscopy (EM).
• Inhibition testing using spiked negative stool samples and Salmon DNA as an internal control.

Main Results:

• EVPrtPCR detected significantly more positive samples (34%) compared to EM (3.8%).
• Norovirus was the most prevalent (17.6%), followed by rotavirus (6.8%), sapovirus (4.2%), astrovirus (2.0%), and enteric adenovirus (1.4%).
• Mixed infections were detected in 9% of samples; EVPrtPCR identified viruses missed by EM, including norovirus and sapovirus.

Conclusions:

• Real-time PCR technology substantially enhances the identification of enteric viruses causing sporadic gastroenteritis.
• The EVPrtPCR panel offers a more sensitive and comprehensive diagnostic approach compared to traditional EM.
• Molecular methods are crucial for accurate etiological diagnosis and understanding the epidemiology of viral gastroenteritis.