caa a22 4500 60615714X CHVBK 20210128100612.0 cr unu---uuuuu 210128e20150401xx s 000 0 eng 10.1007/s10096-014-2284-7 doi (NATIONALLICENCE)springer-10.1007/s10096-014-2284-7 Diagnosis of Clostridium difficile [Elektronische Daten] real-time PCR detection of toxin genes in faecal samples is more sensitive compared to toxigenic culture [M. Jensen, K. Olsen, X. Nielsen, A. Hoegh, R. Dessau, T. Atlung, J. Engberg] The diagnosis of Clostridium difficile infection (CDI) requires the detection of toxigenic C. difficile or its toxins and a clinical assessment. We evaluated the performance of four nucleic acid amplification tests (NAATs) detecting toxigenic C. difficile directly from faeces compared to routine toxigenic culture. In total, 300 faecal samples from Danish hospitalised patients with diarrhoea were included consecutively. Culture was performed in duplicate (routine and ‘expanded toxigenic culture': prolonged and/or re-culture) and genotypic toxin profiling by polymerase chain reaction (PCR), PCR ribotyping and toxinotyping (TT) were performed on culture-positive samples. In parallel, the samples were analysed by four NAATs; two targeting tcdA or tcdB (illumigene® C. difficile and PCRFast® C. difficile A/B) and two multi-target real-time (RT) PCR assays also targeting cdt and tcdC alleles characteristic of epidemic and potentially more virulent PCR ribotypes 027, 066 and 078 (GeneXpert® C. difficile/Epi and an ‘in-house RT PCR' two-step algorithm). The multi-target assays were significantly more sensitive compared to routine toxigenic culture (p < 0.05) and significantly more robust to inhibition compared to PCRFast (p < 0.001). Duplicate ‘expanded toxigenic culture' increased the culture-positive rate by 29% compared to routine culture. The ability of the GeneXpert and in-house assays to correctly classify PCR ribotype 027 was high (>95%), and in-house PCR displayed 100% correct identification of PCR ribotypes 066 and 078. Furthermore, the presence of the PCR enhancer bovine serum albumin (BSA) was found to be related to high sensitivity and low inhibition rate. Rapid laboratory diagnosis of toxigenic C. difficile by RT PCR was accurate. Springer-Verlag Berlin Heidelberg, 2014 Jensen M. Department of Clinical Microbiology, Slagelse Hospital, Ingemannsvej 18, 4200, Slagelse, Denmark aut Olsen K. Department of Microbiology and Infection Control, Statens Serum Institut, Artillerivej 5, 2300, Copenhagen, Denmark aut Nielsen X. Department of Clinical Microbiology, Slagelse Hospital, Ingemannsvej 18, 4200, Slagelse, Denmark aut Hoegh A. Department of Clinical Microbiology, Hvidovre Hospital, Kettegård Allé 30, 2650, Hvidovre, Denmark aut Dessau R. Department of Clinical Microbiology, Slagelse Hospital, Ingemannsvej 18, 4200, Slagelse, Denmark aut Atlung T. Department of Science, Systems and Models, Roskilde University, Universitetsvej 1, 4000, Roskilde, Denmark aut Engberg J. Department of Clinical Microbiology, Slagelse Hospital, Ingemannsvej 18, 4200, Slagelse, Denmark aut European Journal of Clinical Microbiology & Infectious Diseases Springer Berlin Heidelberg 34/4(2015-04-01), 727-736 0934-9723 34:4<727 2015 34 10096 https://doi.org/10.1007/s10096-014-2284-7 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/s10096-014-2284-7 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Jensen M. Department of Clinical Microbiology, Slagelse Hospital, Ingemannsvej 18, 4200, Slagelse, Denmark aut NATIONALLICENCE 700 1- Olsen K. Department of Microbiology and Infection Control, Statens Serum Institut, Artillerivej 5, 2300, Copenhagen, Denmark aut NATIONALLICENCE 700 1- Nielsen X. Department of Clinical Microbiology, Slagelse Hospital, Ingemannsvej 18, 4200, Slagelse, Denmark aut NATIONALLICENCE 700 1- Hoegh A. Department of Clinical Microbiology, Hvidovre Hospital, Kettegård Allé 30, 2650, Hvidovre, Denmark aut NATIONALLICENCE 700 1- Dessau R. Department of Clinical Microbiology, Slagelse Hospital, Ingemannsvej 18, 4200, Slagelse, Denmark aut NATIONALLICENCE 700 1- Atlung T. Department of Science, Systems and Models, Roskilde University, Universitetsvej 1, 4000, Roskilde, Denmark aut NATIONALLICENCE 700 1- Engberg J. Department of Clinical Microbiology, Slagelse Hospital, Ingemannsvej 18, 4200, Slagelse, Denmark aut NATIONALLICENCE 773 0- European Journal of Clinical Microbiology & Infectious Diseases Springer Berlin Heidelberg 34/4(2015-04-01), 727-736 0934-9723 34:4<727 2015 34 10096