Evaluation of a New Multiplex Real-Time PCR Assay for Detecting Gastroenteritis-Causing Viruses in Stool Samples

Area of Science:

• Clinical microbiology
• Molecular diagnostics
• Infectious disease epidemiology

Background:

• Diarrhea is a leading cause of mortality in children under five.
• Rapid and accurate diagnosis of gastrointestinal pathogens is critical for effective treatment and reducing mortality.
• Viral gastroenteritis contributes significantly to childhood morbidity and mortality worldwide.

Purpose of the Study:

• To evaluate the performance of a novel one-step multiplex real-time PCR assay, the Allplex GI-Virus Assay.
• To assess the assay's ability to detect six key diarrhea-causing viruses in stool samples.
• To compare the Allplex assay's diagnostic accuracy against established methods.

Main Methods:

• The Allplex GI-Virus Assay was evaluated using 446 stool samples from patients with acute gastroenteritis.
• Performance was compared to another multiplex PCR assay (Seeplex Diarrhea-V Ace Detection).
• Results were also validated against genotyping by sequencing.

Main Results:

• The Allplex assay demonstrated high overall agreement with the Seeplex assay (93.3%–99.6%) and genotyping (89.7%–99.8%) for tested viruses.
• The assay successfully detected multiple genotypes for rotavirus, norovirus GI and GII, adenovirus, astrovirus, and sapovirus.
• Sapoviruses, not detected by the comparator assay, were identified by the Allplex assay.

Conclusions:

• The Allplex GI-Virus Assay exhibits high concordance with existing diagnostic methods.
• The assay's ability to detect sapoviruses offers an advantage in comprehensive viral gastroenteritis diagnosis.
• This multiplex PCR assay is a valuable tool for identifying viral gastrointestinal infections in clinical settings.