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QUANTITATIVE MULTIPLEX REAL-TIME PCR AS A METHOD FOR DETECTION OF SIGNIFICANT BACTERIURIA IN CHILDREN


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Abstract

Introduction. Urinary tract infections (UTIs) are the second most common of children’s infections, yet current bacteriological methods often are time-consuming and require expensive equipment. Aim of the study is evaluation of diagnostic characteristics of previously developed «AmpliSens® IMP-screen-titer-FL» kit (Central Research Institute of Epidemiology, Moscow) to quantitatively determine bacteriuria as well as the main species of uropathogens in children. Materials and methods. A retrospective analysis of 445 urine samples collected in 2019 from patients of the urological department of the National Medical Research Center of Children’s Health, aged from 4 weeks to 17 years was performed. Urine samples were analyzed with quantitative real-time PCR using the AmpliSens® IMP-screen-titer-FL reagent kit developed earlier for the quantitative detection of significant uropathogens. As a reference method for assessing the diagnostic characteristics of the assay, we used the biomaterial inoculation method on URISELECT™ selective medium (Bio-Rad Laboratories, USA) followed by species identification of microorganisms using the MALDI-TOF MS method (Bruker Daltonics, Germany), as well as Vitek 2 bacterial analyzer (BioMerieux, France). Results. All urine samples were analyzed with significant bacteriuria detected in 122 cases (27%). The most common bacteriuria causing pathogens were Escherichia coli (28%), Klebsiella pneumoniae (18%), and Enterococcus faecalis (14%). Specificity and sensitivity of quantitative real-time PCR for E. coli were 100% and 99%, for K. pneumoniae were 93% and 97%, and for E. faecalis - 91% and 94%, respectively. The sensitivity and specificity for assay previously established in our laboratory were determined as 92% and 95%, respectively. Conclusion. The established real-time quantitative PCR assay is a promising alternative method for determining both the UTI species identification and the titer of bacteria in bacteriuria.


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