Publication Details

Category Text Publication
Reference Category Journals
DOI 10.1371/journal.pone.0197319
Licence creative commons licence
Title (Primary) Molecular quantification and differentiation of Candida species in biological specimens of patients with liver cirrhosis
Author Krohn, S.; Zeller, K.; Böhm, S.; Chatzinotas, A.; Harms, H.; Hartmann, I.; Heidtmann, A.; Herber, A.; Kaiser, T.; Treuheit, M.; Hoffmeister, A.; Berg, T.; Engelmann, C.
Journal PLOS ONE
Year 2018
Department UMB
Volume 13
Issue 6
Page From e0197319
Language englisch
Abstract Patients with liver cirrhosis are susceptible to fungal infections. Due to low sensitivity of culture-based methods, we applied a real-time PCR assay targeting the 18S rRNA gene in combination with direct sequencing and terminal-restriction fragment length polymorphism (T-RFLP) in order to establish a novel tool to detect fungal DNA and to quantify and differentiate Candida DNA, also in polyfungal specimens. In total, 281 samples (blood n = 135, ascites n = 92, duodenal fluid n = 54) from 135 patients with liver cirrhosis and 52 samples (blood n = 26, duodenal fluid n = 26) from 26 control patients were collected prospectively. Candida DNA was quantified in all samples. Standard microbiological culture was performed for comparison. Blood and ascites samples, irrespective of the patient cohort, showed a method-independent low fungal detection rate of approximately 1%, and the Candida DNA content level did not exceed 3.0x101 copies ml-1 in any sample. In contrast, in duodenal fluid of patients with liver cirrhosis high fungal detection rates were discovered by using both PCR- and culture-based techniques (81.5% vs. 66.7%; p = 0.123) and the median level of Candida DNA was 3.8x105 copies ml-1 (2.3x102-6.3x109). In cirrhosis and controls, fungal positive culture results were confirmed by PCR in 96% and an additional amount of 44% of culture negative duodenal samples were PCR positive. Using T-RFLP analysis in duodenal samples, overall 85% of results from microbial culture were confirmed and in 75% of culture-negative but PCR-positive samples additional Candida species could be identified. In conclusion, PCR-based methods and subsequent differentiation of Candida DNA might offer a quick approach to identifying Candida species without prior cultivation.
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Krohn, S., Zeller, K., Böhm, S., Chatzinotas, A., Harms, H., Hartmann, I., Heidtmann, A., Herber, A., Kaiser, T., Treuheit, M., Hoffmeister, A., Berg, T., Engelmann, C. (2018):
Molecular quantification and differentiation of Candida species in biological specimens of patients with liver cirrhosis
PLOS One 13 (6), e0197319