Abstract
Abstract Introduction/Objective Introduction: Standard diagnostic practices for Urinary Tract Infection (UTI) take 2–3 days for pathogen identification and antibiotic susceptibility testing leading to delayed diagnosis and an increase in healthcare expenditures. The rapid diagnostics tool is highly desirable to identify the pathogen and its antibiotic-resistant genes directly from the samples. Our objective is to validate and evaluate the performance of Open array multiplex PCR for pathogen identification and antibiotic resistance genes. Methods/Case Report Method: Open array multiplex PCR method for 20 UTI pathogens was validated using ATCC organisms and inter-laboratory correlation using 50 urine samples. Briefly, samples were homogenized using Omni bead ruptor 96, and DNA was extracted using Hamilton Vantage. Customized open array and UTI chips with 20 most common urinary tract pathogen target and 13 resistant genes target specifically for UTI treatment were filled with Taqman open array PCR master mix and extracted DNA in a 1:1 ratio. UTI chips were prepared on Accufill and real-time PCR was performed on Quantstudio 12K flex. A total of 116 de-identified patient samples were analyzed. Results (if a Case Study enter NA) Result: The sensitivity, specificity, and accuracy of this method were more than 90% considering the reference method as the standard method. A total of 14 different types of organisms were detected. Escherichia coli (n=49) was the highest among all. Multiple pathogens were identified in 27 urine samples. Organisms such as Ureaplasma and Megasphaera 1 were detected in nine and four samples, respectively. All positive samples were resistant to at least one or two resistant genes. tetM and tetS were resistant among 72 samples, erm A, B, and C in 60 samples, ESBL in 30 and fosfo A in 21 samples. Conclusion Conclusion: The open array multiplex PCR technology may help in the early diagnosis and treatment that can improve patient care, shorten hospitalization, and reduces the economic burden.
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