Abstract
Enteric viral pathogens causing gastroenteritis include adenovirus and rotavirus, among others. Rotavirus is the leading cause of severe diarrhea in infants and young children worldwide. Among the adenoviruses known to cause gastroenteritis are those of species F (serotypes 40, 41). Here, we describe the development and validation of a laboratory-developed gastrointestinal triplex rRT-PCR (triplex) assay that targets adenovirus and rotavirus. Stool specimens were tested from patients across Ontario. Specimens were previously tested for adenovirus and/or rotavirus by electron microscopy (EM) or immunochromatographic test (ICT). Triplex sensitivity, specificity, positive and negative predictive values compared to Seegene assay (a commercial assay used here as the standard reference method) were 100%, 97.8%, 86.0%, 100% for adenovirus, and 99.1%, 98.4%, 96.3%. 99.6% for rotavirus, respectively. The triplex assay had a 95.2% and 97.3% overall percent agreements (OPAs) when compared to EM for adenovirus or rotavirus detection, respectively, and an OPA of 90.9% when compared to rotavirus ICT for rotavirus detection. Triplex assay exhibited similar performance to the Seegene assay for both adenovirus and rotavirus and detected more adenovirus and rotavirus than traditional testing methods. The high performance along with lower cost and reduced turnaround time makes the triplex assay a desirable testing method for a clinical microbiology laboratory.
Highlights
Infectious diarrhea is the second most common infectious condition worldwide, after acute respiratory infections, affecting humans of all age groups, but mostly the young, the elderly, the immunocompromised patients and people in enclosed communities such as hospitals, nursing homes, military bases and cruise ships [1,2,3]
This study utilized clinical stool specimens submitted to Public Health Ontario (PHO) laboratory for routine gastrointestinal virus testing, including adenovirus and rotavirus, from patients across Ontario
For the purpose of this validation, positive specimens for adenovirus or rotavirus refer to those previously confirmed positive by electron microscopy (EM) and/or immunochromatographic test (ICT) tests
Summary
Infectious diarrhea is the second most common infectious condition worldwide, after acute respiratory infections, affecting humans of all age groups, but mostly the young, the elderly, the immunocompromised patients and people in enclosed communities such as hospitals, nursing homes, military bases and cruise ships [1,2,3]. As causative agents of gastroenteritis, include adenovirus, rotavirus, norovirus, sapovirus and astrovirus [4,5,6]. Several bacterial pathogens, such as Campylobacter jejuni, enterotoxigenic Escherichia Coli, Shigella species and Vibrio cholera, are reported to cause gastroenteritis, in developing countries and refugee camps [7]. Rotavirus was the leading pathogen causing diarrhea in infants and young children in Pathogens 2020, 9, 326; doi:10.3390/pathogens9050326 www.mdpi.com/journal/pathogens
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