Comparing single versus multiple virus detection in pediatric acute gastroenteritis postimplementation of routine multiplex RT‐PCR diagnostic testing

Abstract

Utilizing multiplex real time polymerase chain reaction (RT‐PCR) for rapid diagnosis of gastroenteritis, enables simultaneous detection of multiple pathogens. A comparative analysis of disease characteristics was conducted between cases with single and multiple viruses. Rotavirus vaccine was introduced in 2010, reaching a 70% coverage in 2 years. All rectal swabs collected from diarrheic children (<5 years) between December 2017 and March 2022 were included. Detection of the same viruses within 2 months was considered a single episode. Episodes with positive stool bacterial PCR were excluded. A total of 5879 samples were collected, revealing 86.9% (1509) with single virus detection and 13.1% (227) with multiple viruses. The most frequent combination was rotavirus and norovirus (27.8%), these infections followed a winter‐spring seasonality akin to rotavirus. Children with multivirus infections exhibited higher immunodeficiency (OR 2.06) rates, but lower food allergy (OR 0.45) and prematurity rates (OR 0.55) compared to single infections. Greater disease severity, evaluated by the Vesikari score, was observed in multivirus episodes (p < 0.001, OR 1.12). Multivirus infections accounted for 13.1% of symptomatic cases in hospitalized young children. Despite vaccination efforts, rotavirus remained prominent, frequently in co‐infections with norovirus. Overall, multivirus infections were linked to more severe diseases than single virus cases.

Risk factors and clinical outcomes associated with multiple as opposed to single pathogens detected on the gastrointestinal disease polymerase chain reaction assay

“…Prior studies of viral diarrhea in children have suggested that there is greater severity of diarrhea when multiple viruses are detected, but less is known in adults and with bacterial enteropathogens. 31 After adjusting for other factors including age, insurance status, and immunosuppression, detection of multiple pathogens was associated with a 44% increased risk for subsequent ED visits compared to detection of a single pathogen. Those with multiple pathogens were also more likely to receive more than one antibiotic, although overall rates of antibiotic use were similar comparing those with multiple vs. single pathogens.…”

“…[26][27][28][29] Hispanic ethnicity may be associated with the detection of multiple pathogens on GI PCR due to a combination of host genetics, geographic location, and socioeconomic factors such as diet, living environment, pathogen exposures, access to medical care, travel, and other social constructs that shape the gut microbiome and influence susceptibility to enteric pathogens. [30][31][32][33][34][35][36][37][38] This study could not determine the specific factors underlying the association between Hispanic ethnicity and higher incidence of multiple pathogens on GI PCR testing.…”

Enterovirus genotype diversity with emergence of coxsackievirus A2 circulating in pediatric patients with acute gastroenteritis in Thailand, 2019–2022

“…Studies from the Netherlands ( Pijnacker et al, 2019 ), Japan ( Hikita et al, 2023 ), and Nepal ( Tandukar et al, 2019 ) demonstrated that co-infection did not increase the risk or severity of AGE. Conversely, a study conducted in Israel indicated that co-infection was associated with more severe clinical symptoms in children under five years old ( Hazan et al, 2024 ). Although the impact of co-infection on the severity of AGE remains unclear, the detection of viral load in stool samples using real-time PCR could be useful in assessing the contribution of individual viruses to the severity of disease as well as virus-virus interaction in co-infection cases ( De Grazia et al, 2020 ).…”