Application of metagenomic next-generation sequencing in the diagnosis of urinary tract infection in patients undergoing cutaneous ureterostomy

Abstract

ObjectiveUrinary tract infection (UTI) is an inflammatory response of the urothelium to bacterial invasion and is a common complication in patients with cutaneous ureterostomy (CU). For such patients, accurate and efficient identification of pathogens remains a challenge. The aim of this study included exploring utility of metagenomic next-generation sequencing (mNGS) in assisting microbiological diagnosis of UTI among patients undergoing CU, identifying promising cytokine or microorganism biomarkers, revealing microbiome diversity change and compare virulence factors (VFs) and antibiotic resistance genes (ARGs) after infection.MethodsWe performed a case-control study of 50 consecutive CU patients from December 2020 to January 2021. According to the clinical diagnostic criteria, samples were divided into infected group and uninfected group and difference of urine culture, cytokines, microorganism, ARGs and VFs were compared between the two groups.ResultsInflammatory responses were more serious in infected group, as evidenced by a significant increase in IFN-α (p=0.031), IL-1β (0.023) and IL-6 (p=0.018). Clinical culture shows that there is higher positive rate in infected group for most clinical pathogens like Escherichia coli, Klebsiella pneumoniae, Staphylococcus aureus, Candida auris etc. and the top three pathogens with positive frequencies were E. coli, K. pneumoniae, and Enterococcus faecalis. Benchmarking clinical culture, the total sensitivity is 91.4% and specificity is 76.3% for mNGS. As for mNGS, there was no significant difference in microbiome α- diversity between infected and uninfected group. Three species biomarkers including Citrobacter freundii, Klebsiella oxytoca, and Enterobacter cloacae are enriched in infected group based on Lefse. E. cloacae were significantly correlated with IL-6 and IL-10. K. oxytoca were significantly correlated with IL-1β. Besides, the unweighted gene number and weighted gene abundance of VFs or ARGs are significantly higher in infected group. Notablely, ARGs belonging to fluoroquinolones, betalatmas, fosfomycin, phenicol, phenolic compound abundance is significantly higher in infected group which may have bad effect on clinical treatment for patients.ConclusionmNGS, along with urine culture, will provide comprehensive and efficient reference for the diagnosis of UTI in patients with CU and allow us to monitor microbial changes in urine of these patients. Moreover, cytokines (IL-6, IL-1β, and IFN-a) or microorganisms like C. freundii, K. oxytoca or E. cloacae are promising biomarkers for building effective UTI diagnostic model of patients with CU and seriously the VFs and ARGs abundance increase in infected group may play bad effect on clinical treatment.

Analytical Validation of a Highly Accurate and Reliable Next-Generation Sequencing-Based Urine Assay

“…Regarding concerns about the potential oversensitivity of an NGS-based assay, BIOTIA-DX was intentionally designed and trained to increase stringency and reduce false positive detection of urogenital commensals or opportunistic pathogens present at colonization levels often a common weakness of many molecular diagnostic tools and clinical studies (34,81,88,100,101). Relative abundance of microbial species, percentage of human reads, IPC detection and complexity of the microbial profiles are features implemented to enhance the BIOTIA-DX's ability to correctly predict an infection.…”

Uropathogenic Escherichia coli infection: innate immune disorder, bladder damage, and Tailin Fang II

“…And subsequently, they should play significant roles in the body’s proinflammatory and anti-inflammatory responses through interactions with a variety of receptors ( Behzadi et al., 2022 ). IL-6 and IL-1β are commonly utilized as markers to detect inflammation levels in UTIs, and their increased levels indicate a higher severity of UTI ( Ching et al., 2018 ; Huang et al., 2023 ).…”

Diversity and antibiotic resistance profile analysis of uropathogenic bacteria in human and canines

“…Urinary tract infection (UTI) is considered as most prevalent and second most common type of infection in the body. The infection is caused by the invasion of pathogens in the urinary tract and produces the inflammatory response of urothelium [11,12]. Main cause of urinary tract infection is the proliferation of bacteria in urinary tract.…”