caa a22 4500 606155740 CHVBK 20210128100605.0 cr unu---uuuuu 210128e20150901xx s 000 0 eng 10.1007/s10096-015-2425-7 doi (NATIONALLICENCE)springer-10.1007/s10096-015-2425-7 Fecal detection of enterotoxigenic Bacteroides fragilis [Elektronische Daten] [L. Chen, S. Van Meerbeke, E. Albesiano, A. Goodwin, S. Wu, H. Yu, K. Carroll, C. Sears] Bacteroides fragilis is a common colonic symbiote of which one subtype, enterotoxigenic Bacteroides fragilis (ETBF), causes inflammatory diarrhea. However, asymptomatic ETBF colonization is common. Through its primary virulence factor, B. fragilis toxin (BFT), ETBF causes asymptomatic, chronic colitis in C57BL/6 mice and increased colon tumorigenesis in multiple intestinal neoplasia mice. Human studies suggest an association between ETBF infection, inflammatory bowel disease, and colon cancer. Additional studies on ETBF epidemiology are, therefore, crucial. The goal of this study is to develop a reliable fecal diagnostic for ETBF. To develop a sensitive assay for ETBF, we tested multiple protocols on mouse stools spiked with serially diluted ETBF. Each assay was based on either touchdown or quantitative polymerase chain reaction (qPCR) and used primers targeted to bft to detect ETBF. Using touchdown PCR or qPCR, the mean ETBF detection limit was 1.55 × 106 colony-forming units (CFU)/g stool and 1.33 × 104CFU/g stool, respectively. Augmentation of Bacteroides spp. growth in fecal samples using PYGB (Peptone Yeast Glucose with Bile) broth enhanced ETBF detection to 2.93 × 102CFU/g stool using the touchdown PCR method and 2.63 × 102CFU/g stool using the qPCR method. Fecal testing using combined culture-based amplification and bft touchdown PCR is a sensitive assay for the detection of ETBF colonization and should be useful in studying the role of ETBF colonization in intestinal diseases, such as inflammatory bowel disease and colon cancer. We conclude that touchdown PCR with culture-based amplification may be the optimal ETBF detection strategy, as it performs as well as qPCR with culture-based amplification, but is a less expensive technique. Springer-Verlag Berlin Heidelberg, 2015 Chen L. Department of Medicine, Johns Hopkins School of Medicine, CRB2 Bldg, Suite 1M.05, 1550 Orleans Street, 21231, Baltimore, MD, USA aut Van Meerbeke S. Department of Medicine, Johns Hopkins School of Medicine, CRB2 Bldg, Suite 1M.05, 1550 Orleans Street, 21231, Baltimore, MD, USA aut Albesiano E. Department of Medicine, Johns Hopkins School of Medicine, CRB2 Bldg, Suite 1M.05, 1550 Orleans Street, 21231, Baltimore, MD, USA aut Goodwin A. Department of Medicine, Johns Hopkins School of Medicine, CRB2 Bldg, Suite 1M.05, 1550 Orleans Street, 21231, Baltimore, MD, USA aut Wu S. Department of Medicine, Johns Hopkins School of Medicine, CRB2 Bldg, Suite 1M.05, 1550 Orleans Street, 21231, Baltimore, MD, USA aut Yu H. Sidney Kimmel Comprehensive Cancer Center, Department of Oncology, Johns Hopkins School of Medicine, CRB1 1650 Orleans Street, 21287, Baltimore, MD, USA aut Carroll K. Department of Pathology, Johns Hopkins School of Medicine, Meyer B1-193, 600 North Wolfe Street, 21287, Baltimore, MD, USA aut Sears C. Department of Medicine, Johns Hopkins School of Medicine, CRB2 Bldg, Suite 1M.05, 1550 Orleans Street, 21231, Baltimore, MD, USA aut European Journal of Clinical Microbiology & Infectious Diseases Springer Berlin Heidelberg 34/9(2015-09-01), 1871-1877 0934-9723 34:9<1871 2015 34 10096 https://doi.org/10.1007/s10096-015-2425-7 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/s10096-015-2425-7 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Chen L. Department of Medicine, Johns Hopkins School of Medicine, CRB2 Bldg, Suite 1M.05, 1550 Orleans Street, 21231, Baltimore, MD, USA aut NATIONALLICENCE 700 1- Van Meerbeke S. Department of Medicine, Johns Hopkins School of Medicine, CRB2 Bldg, Suite 1M.05, 1550 Orleans Street, 21231, Baltimore, MD, USA aut NATIONALLICENCE 700 1- Albesiano E. Department of Medicine, Johns Hopkins School of Medicine, CRB2 Bldg, Suite 1M.05, 1550 Orleans Street, 21231, Baltimore, MD, USA aut NATIONALLICENCE 700 1- Goodwin A. Department of Medicine, Johns Hopkins School of Medicine, CRB2 Bldg, Suite 1M.05, 1550 Orleans Street, 21231, Baltimore, MD, USA aut NATIONALLICENCE 700 1- Wu S. Department of Medicine, Johns Hopkins School of Medicine, CRB2 Bldg, Suite 1M.05, 1550 Orleans Street, 21231, Baltimore, MD, USA aut NATIONALLICENCE 700 1- Yu H. Sidney Kimmel Comprehensive Cancer Center, Department of Oncology, Johns Hopkins School of Medicine, CRB1 1650 Orleans Street, 21287, Baltimore, MD, USA aut NATIONALLICENCE 700 1- Carroll K. Department of Pathology, Johns Hopkins School of Medicine, Meyer B1-193, 600 North Wolfe Street, 21287, Baltimore, MD, USA aut NATIONALLICENCE 700 1- Sears C. Department of Medicine, Johns Hopkins School of Medicine, CRB2 Bldg, Suite 1M.05, 1550 Orleans Street, 21231, Baltimore, MD, USA aut NATIONALLICENCE 773 0- European Journal of Clinical Microbiology & Infectious Diseases Springer Berlin Heidelberg 34/9(2015-09-01), 1871-1877 0934-9723 34:9<1871 2015 34 10096