caa a22 4500 606156518 CHVBK 20210128100609.0 cr unu---uuuuu 210128e20150301xx s 000 0 eng 10.1007/s10096-014-2257-x doi (NATIONALLICENCE)springer-10.1007/s10096-014-2257-x Evaluation of a real-time multiplex PCR for the simultaneous detection of Campylobacter jejuni , Salmonella spp., Shigella spp./EIEC, and Yersinia enterocolitica in fecal samples [Elektronische Daten] [P. Van Lint, E. De Witte, H. De Henau, A. De Muynck, L. Verstraeten, B. Van Herendael, S. Weekx] Conventional diagnosis of infectious diarrhea caused by bacteria is time-consuming, labor-intensive, and has a suboptimal sensitivity. We have therefore developed a multiplex real-time polymerase chain reaction (PCR) for the simultaneous detection of Campylobacter jejuni, Salmonella spp., Shigella spp./enteroinvasive Escherichia coli (EIEC), and Yersinia enterocolitica in fecal samples. No cross reactivity between the different pathogens was observed, and the multiplex setup of the assay did not have an impact on the sensitivity of the PCR. The analytical sensitivity was 87CFU/mL for C. jejuni, 61CFU/mL for Shigella spp./EIEC, 5,528CFU/mL for Salmonella spp., and 1,306CFU/mL for Y. enterocolitica. An extensive validation of the assay was performed by testing 1,687 patient samples by both PCR and with conventional techniques. The use of PCR increased the overall clinical sensitivity from 78 to 100% (p < 0.0001), the specificity was 99.4% for the PCR, compared with 99.9% for conventional culture. The novel PCR assay allows for rapid, sensitive, inexpensive, and high-throughput testing of the most common bacterial causes of gastroenteritis. Springer-Verlag Berlin Heidelberg, 2014 Van Lint P. Clinical Laboratory GZA, Department of Molecular Diagnostics, GZA St-Augustinus, Oosterveldlaan 24, 2610, Wilrijk, Belgium aut De Witte E. Clinical Laboratory GZA, Department of Molecular Diagnostics, GZA St-Augustinus, Oosterveldlaan 24, 2610, Wilrijk, Belgium aut De Henau H. Clinical Laboratory GZA, Department of Molecular Diagnostics, GZA St-Augustinus, Oosterveldlaan 24, 2610, Wilrijk, Belgium aut De Muynck A. Clinical Laboratory GZA, Department of Molecular Diagnostics, GZA St-Augustinus, Oosterveldlaan 24, 2610, Wilrijk, Belgium aut Verstraeten L. Clinical Laboratory GZA, Department of Molecular Diagnostics, GZA St-Augustinus, Oosterveldlaan 24, 2610, Wilrijk, Belgium aut Van Herendael B. Clinical Laboratory GZA, Department of Molecular Diagnostics, GZA St-Augustinus, Oosterveldlaan 24, 2610, Wilrijk, Belgium aut Weekx S. Clinical Laboratory GZA, Department of Molecular Diagnostics, GZA St-Augustinus, Oosterveldlaan 24, 2610, Wilrijk, Belgium aut European Journal of Clinical Microbiology & Infectious Diseases Springer Berlin Heidelberg 34/3(2015-03-01), 535-542 0934-9723 34:3<535 2015 34 10096 https://doi.org/10.1007/s10096-014-2257-x text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s10096-014-2257-x text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Van Lint P. Clinical Laboratory GZA, Department of Molecular Diagnostics, GZA St-Augustinus, Oosterveldlaan 24, 2610, Wilrijk, Belgium aut NATIONALLICENCE 700 1- De Witte E. Clinical Laboratory GZA, Department of Molecular Diagnostics, GZA St-Augustinus, Oosterveldlaan 24, 2610, Wilrijk, Belgium aut NATIONALLICENCE 700 1- De Henau H. Clinical Laboratory GZA, Department of Molecular Diagnostics, GZA St-Augustinus, Oosterveldlaan 24, 2610, Wilrijk, Belgium aut NATIONALLICENCE 700 1- De Muynck A. Clinical Laboratory GZA, Department of Molecular Diagnostics, GZA St-Augustinus, Oosterveldlaan 24, 2610, Wilrijk, Belgium aut NATIONALLICENCE 700 1- Verstraeten L. Clinical Laboratory GZA, Department of Molecular Diagnostics, GZA St-Augustinus, Oosterveldlaan 24, 2610, Wilrijk, Belgium aut NATIONALLICENCE 700 1- Van Herendael B. Clinical Laboratory GZA, Department of Molecular Diagnostics, GZA St-Augustinus, Oosterveldlaan 24, 2610, Wilrijk, Belgium aut NATIONALLICENCE 700 1- Weekx S. Clinical Laboratory GZA, Department of Molecular Diagnostics, GZA St-Augustinus, Oosterveldlaan 24, 2610, Wilrijk, Belgium aut NATIONALLICENCE 773 0- European Journal of Clinical Microbiology & Infectious Diseases Springer Berlin Heidelberg 34/3(2015-03-01), 535-542 0934-9723 34:3<535 2015 34 10096