Abstract
Many microbial species have been recognized as enteropathogens for humans. Here, we predicted the causative agents of acute diarrhea using data from multiplex quantitative PCR (qPCR) assays targeting 19 enteropathogens. For this, a case-control study was conducted at eight hospitals in Thailand. Stool samples and clinical data were collected from 370 hospitalized patients with acute diarrhea and 370 non-diarrheal controls. Multiple enteropathogens were detected in 75.7% and 13.0% of diarrheal stool samples using multiplex qPCR and bacterial culture methods, respectively. Asymptomatic carriers of enteropathogens were found among 87.8% and 45.7% of individuals by qPCR and culture methods, respectively. These results suggested the complexity of identifying causative agents of diarrhea. An analysis using the quantification cut-off values for clinical relevance drastically reduced pathogen-positive stool samples in control subjects from 87.8% to 0.5%, whereas 48.9% of the diarrheal stool samples were positive for any of the 11 pathogens. Among others, rotavirus, norovirus GII, Shigella/EIEC, and Campylobacter were strongly associated with acute diarrhea (P-value < 0.001). Characteristic clinical symptoms, epidemic periods, and age-related susceptibility to infection were observed for some enteropathogens. Investigations based on qPCR approaches covering a broad array of enteropathogens might thus improve our understanding of diarrheal disease etiology and epidemiological trends.
Highlights
Many microbial species have been recognized as enteropathogens for humans
With the clinically relevant cut-off values, pathogen-positive stool samples in control subjects were drastically reduced from 87.8% to 0.5%, whereas 181 diarrheal stools (48.9%) were positive for any of the 11 enteropathogens - rotavirus, norovirus GII, Shigella/enteroinvasive E. coli (EIEC), Campylobacter, enteroaggregative E. coli (EAEC), Salmonella, Plesiomonas, Aeromonas, sapovirus, astrovirus, and enterotoxigenic E. coli (ETEC) – that were strongly associated with acute diarrhea (Fig. 2B, Table 1)
We investigated the etiological agents of acute diarrhea in patients with severe symptoms by quantitatively detecting a broad range of known enteropathogens
Summary
We predicted the causative agents of acute diarrhea using data from multiplex quantitative PCR (qPCR) assays targeting 19 enteropathogens. Multiple enteropathogens were detected in 75.7% and 13.0% of diarrheal stool samples using multiplex qPCR and bacterial culture methods, respectively. Asymptomatic carriers of enteropathogens were found among 87.8% and 45.7% of individuals by qPCR and culture methods, respectively These results suggested the complexity of identifying causative agents of diarrhea. Molecular assays targeting nucleic acid markers facilitate the screening of a broad range of enteropathogens in stool samples, consuming much less time than conventional methods such as cell culture, microscopy, and antigen-based tests. We predicted the etiological agents of acute diarrhea using stool samples from hospitalized patients in Thailand using our quantitative pathogen detection procedure[22] with clinically relevant cut-off values. We further investigated characteristic clinical symptoms, epidemic periods, and age-related susceptibility to infection associated with the detected enteropathogens
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