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Articles by Urban Ryberg in JoVE
JoVE General
Productie van Weefsel Microarrays, Immunohistochemie vlekken en Digitalisering Binnen het menselijke eiwit Atlas
Department of Immunology, Genetics and Pathology, Science for Life Laboratory, Uppsala University
Weefsel microarrays zorgt voor een efficiënte methode om gelijktijdige informatie te verkrijgen uit een veelheid van weefsels. Representatieve delen van weefsels worden samengevoegd tot een paraffineblok. Secties van het blok voor immunohistochemie en analyse van eiwit expressie. Digitaal scannen genereert corresponderende beelden voor de distributie van gegevens.
Other articles by Urban Ryberg on PubMed
High-resolution Melting-curve Analysis of Ligation-mediated Real-time PCR for Rapid Evaluation of an Epidemiological Outbreak of Extended-spectrum-beta-lactamase-producing Escherichia Coli
Methods for the confirmation of nosocomial outbreaks of bacterial pathogens are complex, expensive, and time-consuming. Recently, a method based on ligation-mediated PCR (LM/PCR) using a low denaturation temperature which produces specific melting-profile patterns of DNA products has been described. Our objective was to further develop this method for real-time PCR and high-resolution melting analysis (HRM) in a single-tube system optimized in order to achieve results within 1 day. Following the optimization of LM/PCR for real-time PCR and HRM (LM/HRM), the method was applied for a nosocomial outbreak of extended-spectrum-beta-lactamase (ESBL)-producing and ST131-associated Escherichia coli isolates (n = 15) and control isolates (n = 29), including four previous clusters. The results from LM/HRM were compared to results from pulsed-field gel electrophoresis (PFGE), which served as the gold standard. All isolates from the nosocomial outbreak clustered by LM/HRM, which was confirmed by gel electrophoresis of the LM/PCR products and PFGE. Control isolates that clustered by LM/PCR (n = 4) but not by PFGE were resolved by confirmatory gel electrophoresis. We conclude that LM/HRM is a rapid method for the detection of nosocomial outbreaks of bacterial infections caused by ESBL-producing E. coli strains. It allows the analysis of isolates in a single-tube system within a day, and the discriminatory power is comparable to that of PFGE.
