Skip to main content
SpringerLink
Log in

Presence of multiple bacterial markers in clinical samples might be useful for presumptive diagnosis of infection in cirrhotic patients with culture-negative reports

  • Original Article
  • Published:
European Journal of Clinical Microbiology & Infectious Diseases Aims and scope Submit manuscript

Abstract

Bacterial infections in cirrhotic patients with ascites are associated with a severe prognosis and an increased risk of death. The microbiological standard tests for the diagnosis of suspected infection, based on culture test of blood and ascitic fluid, are, in many cases (30–40 %), negative, even when patients show symptoms of infection. A multiple culture-independent protocol was applied and evaluated as a diagnostic and prognostic tool for the detection of bacterial infection in cirrhotic patients. Sixty-four culture-negative samples obtained from 34 cirrhotic patients, with PMN < 250 cells/μl of ascitic fluid, were screened for the presence of bacterial DNA, endotoxin, peptidoglycan/β-glucan and microscopically visible bacterial cells. Correlations between the presence of multiple markers and various clinical and laboratory parameters were evaluated. Bacterial DNA was detected in 23 samples collected from 16 patients; a large part of these samples also showed the presence of other bacterial markers, which was associated with a worsening of liver functionality, a higher incidence of infections during the follow-up and a higher mortality rate in our cohort of cirrhotic patients. We believe that the detection of additional bacterial markers in bacterial DNA-positive clinical samples makes the bacterial presence and its clinical significance more realistic and might be useful as early markers of an ongoing bacterial infection and in establishing a clinical prognosis.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1

Similar content being viewed by others

References

  1. Fernández J, Navasa M, Gómez J, Colmenero J, Vila J, Arroyo V et al (2002) Bacterial infections in cirrhosis: epidemiological changes with invasive procedures and norfloxacin prophylaxis. Hepatology 35:140–148. doi:10.1053/jhep.2002.30082

    Article  PubMed  Google Scholar 

  2. Zapater P, Francés R, González-Navajas JM, de la Hoz MA, Moreu R, Pascual S et al (2008) Serum and ascitic fluid bacterial DNA: a new independent prognostic factor in noninfected patients with cirrhosis. Hepatology 48:1924–1931. doi:10.1002/hep.22564

    Article  CAS  PubMed  Google Scholar 

  3. Merli M, Lucidi C, Giannelli V, Giusto M, Riggio O, Falcone M, Ridola L et al (2010) Cirrhotic patients are at risk for health care-associated bacterial infections. Clin Gastroenterol Hepatol 8(11):979–985. doi:10.1016/j.cgh.2010.06.024

    Article  PubMed  Google Scholar 

  4. Arvaniti V, D’Amico G, Fede G, Manousou P, Tsochatzis E, Pleguezuelo M et al (2010) Infections in patients with cirrhosis increase mortality four-fold and should be used in determining prognosis. Gastroenterology 139:1246–1256. doi:10.1053/j.gastro.2010.06.019

    Article  PubMed  Google Scholar 

  5. Berg RD, Garlington AW (1979) Translocation of certain indigenous bacteria from the gastrointestinal tract to the mesenteric lymph nodes and other organs in a gnotobiotic mouse model. Infect Immun 23:403–411

    PubMed Central  CAS  PubMed  Google Scholar 

  6. Such J, Francés R, Muñoz C, Zapater P, Casellas JA, Cifuentes A et al (2002) Detection and identification of bacterial DNA in patients with cirrhosis and culture-negative, nonneutrocytic ascites. Hepatology 36:135–141. doi:10.1053/jhep.2002.33715

    Article  CAS  PubMed  Google Scholar 

  7. Cirera I, Bauer TM, Navasa M, Vila J, Grande L, Taurá P et al (2001) Bacterial translocation of enteric organisms in patients with cirrhosis. J Hepatol 34:32–37

    Article  CAS  PubMed  Google Scholar 

  8. Jalan R, Fernandez J, Wiest R, Schnabl B, Moreau R, Angeli P et al (2014) Bacterial infections in cirrhosis: a position statement based on the EASL Special Conference 2013. J Hepatol 60(6):1310–1324

    Article  PubMed  Google Scholar 

  9. Thalheimer U, De Iorio F, Capra F, del Mar Lleo M, Zuliani V, Ghidini V et al (2010) Altered intestinal function precedes the appearance of bacterial DNA in serum and ascites in patients with cirrhosis: a pilot study. Eur J Gastroenterol Hepatol 22(10):1228–1234. doi:10.1097/MEG.0b013e32833b4b03

    Article  PubMed  Google Scholar 

  10. Wiest R, Lawson M, Geuking M (2014) Pathological bacterial translocation in liver cirrhosis. J Hepatol 60(1):197–209

    Article  PubMed  Google Scholar 

  11. Bellot P, García-Pagán JC, Francés R, Abraldes JG, Navasa M, Pérez-Mateo M et al (2010) Bacterial DNA translocation is associated with systemic circulatory abnormalities and intrahepatic endothelial dysfunction in patients with cirrhosis. Hepatology 52(6):2044–2052. doi:10.1002/hep.23918

    Article  CAS  PubMed  Google Scholar 

  12. Bellot P, Francés R, Such J (2013) Pathological bacterial translocation in cirrhosis: pathophysiology, diagnosis and clinical implications. Liver Int 33(1):31–39. doi:10.1111/liv.12021

    Article  CAS  PubMed  Google Scholar 

  13. Caño R, Llanos L, Zapater P, Pascual S, Bellot P, Barquero C et al (2010) Proteomic evidence of bacterial peptide translocation in afebrile patients with cirrhosis and ascites. J Mol Med (Berl) 88:487–495. doi:10.1007/s00109-009-0582-9

    Article  Google Scholar 

  14. Borzio M, Salerno F, Piantoni L, Cazzaniga M, Angeli P, Bissoli F et al (2001) Bacterial infection in patients with advanced cirrhosis: a multicentre prospective study. Dig Liver Dis 33(1):41–48

    Article  CAS  PubMed  Google Scholar 

  15. Francés R, Benlloch S, Zapater P, González JM, Lozano B, Muñoz C et al (2004) A sequential study of serum bacterial DNA in patients with advanced cirrhosis and ascites. Hepatology 39:484–491. doi:10.1002/hep.20055

    Article  PubMed  Google Scholar 

  16. Bruns T, Sachse S, Straube E, Assefa S, Herrmann A, Hagel S et al (2009) Identification of bacterial DNA in neutrocytic and non-neutrocytic cirrhotic ascites by means of a multiplex polymerase chain reaction. Liver Int 29:1206–1214. doi:10.1111/j.1478-3231.2009.02073.x

    Article  CAS  PubMed  Google Scholar 

  17. Weisburg WG, Barns SM, Pelletier DA, Lane DJ (1991) 16S ribosomal DNA amplification for phylogenetic study. J Bacteriol 173:697–703

    PubMed Central  CAS  PubMed  Google Scholar 

  18. Muyzer G, de Waal EC, Uitterlinden AG (1993) Profiling of complex microbial populations by denaturing gradient gel electrophoresis analysis of polymerase chain reaction-amplified genes coding for 16S rRNA. Appl Environ Microbiol 59(3):695–700

    PubMed Central  CAS  PubMed  Google Scholar 

  19. del Mar Lleó M, Tafi MC, Signoretto C, Dal Cero C, Canepari P (1999) Competitive polymerase chain reaction for quantification of nonculturable Enterococcus faecalis cells in lake water. FEMS Microbiol Ecol 30:345–353

    Article  PubMed  Google Scholar 

  20. Lleo MM, Bonato B, Tafi MC, Signoretto C, Pruzzo C, Canepari P (2005) Molecular vs culture methods for the detection of bacterial faecal indicators in groundwater for human use. Lett Appl Microbiol 40(4):289–294. doi:10.1111/j.1472-765X.2005.01666.x

    Article  CAS  PubMed  Google Scholar 

  21. Berney M, Hammes F, Bosshard F, Weilenmann HU, Egli T (2007) Assessment and interpretation of bacterial viability by using the LIVE/DEAD BacLight Kit in combination with flow cytometry. Appl Environ Microbiol 73(10):3283–3290. doi:10.1128/AEM.02750-06

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  22. Fagan KJ, Rogers GB, Melino M, Arthur DM, Costello ME, Morrison M et al (2015) Ascites bacterial burden and immune cell profile are associated with poor clinical outcomes in the absence of overt infection. PLoS One 10(3):e0120642. doi:10.1371/journal.pone.0120642

    Article  PubMed Central  PubMed  Google Scholar 

  23. Rampini SK, Bloemberg GV, Keller PM, Büchler AC, Dollenmaier G, Speck RF et al (2011) Broad-range 16S rRNA gene polymerase chain reaction for diagnosis of culture-negative bacterial infections. Clin Infect Dis 53:1245–1251. doi:10.1093/cid/cir692

    Article  CAS  PubMed  Google Scholar 

  24. Lleo MM, Ghidini V, Tafi MC, Castellani F, Trento I, Boaretti M (2014) Detecting the presence of bacterial DNA by PCR can be useful in diagnosing culture-negative cases of infection, especially in patients with suspected infection and antibiotic therapy. FEMS Microbiol Lett 354(2):153–160. doi:10.1111/1574-6968.12422

    Article  CAS  PubMed  Google Scholar 

  25. Francés R, Zapater P, González-Navajas JM, Muñoz C, Caño R, Moreu R et al (2008) Bacterial DNA in patients with cirrhosis and noninfected ascites mimics the soluble immune response established in patients with spontaneous bacterial peritonitis. Hepatology 47(3):979–985. doi:10.1002/hep.22083

    Article  Google Scholar 

  26. Rogers GB, Russell LE, Preston PG, Marsh P, Collins JE, Saunders J et al (2010) Characterisation of bacteria in ascites—reporting the potential of culture-independent, molecular analysis. Eur J Clin Microbiol Infect Dis 29:533–541. doi:10.1007/s10096-010-0891-5

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Diagnostics and Public Health, Section of Microbiology, University of Verona, Strada Le Grazie, 8, 37134, Verona, Italy

    M. Boaretti, F. Castellani & M. M. Lleo

  2. Division of Gastroenterology, Department of Clinical Medicine, “Sapienza” University of Rome, Rome, Italy

    M. Merli & C. Lucidi

Authors
  1. M. Boaretti
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. F. Castellani
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. M. Merli
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. C. Lucidi
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. M. M. Lleo
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to M. M. Lleo.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Boaretti, M., Castellani, F., Merli, M. et al. Presence of multiple bacterial markers in clinical samples might be useful for presumptive diagnosis of infection in cirrhotic patients with culture-negative reports. Eur J Clin Microbiol Infect Dis 35, 433–441 (2016). https://doi.org/10.1007/s10096-015-2556-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10096-015-2556-x

Keywords

Search

Navigation