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Published in: European Journal of Clinical Microbiology & Infectious Diseases 7/2012

Open Access 01-07-2012 | Article

Panbacterial real-time PCR to evaluate bacterial burden in chronic wounds treated with Cutimed™ Sorbact™

Authors: V. Gentili, S. Gianesini, P. G. Balboni, E. Menegatti, A. Rotola, M. Zuolo, E. Caselli, P. Zamboni, D. Di Luca

Published in: European Journal of Clinical Microbiology & Infectious Diseases | Issue 7/2012

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Abstract

The impact of polymicrobial bacterial infection on chronic wounds has been studied extensively, but standard bacteriological analysis is not always sensitive enough. Molecular approaches represent a promising alternative to the standard bacteriological analysis. This work aimed to assess the usefulness of a panbacterial quantitative real-time PCR reaction to quantitate the total bacterial load in chronic wounds treated with Cutimed™ Sorbact™, a novel therapeutic approach based on hydrophobic binding of bacteria to a membrane. The results obtained by panbacterial real-time PCR on conserved sequences of the bacterial 16S gene show that the bacterial burden significantly decreased in 10 out of 15 healing chronic wounds, and did not change in 5 out of 5 non-healing chronic wounds. On the contrary, classical culture for S. aureus and P. aeruginosa, and real-time PCR for Bacteroides and Fusobacterium did not show any correlation with the clinical outcome. Our study also shows that quantification of chronic wounds by panbacterial real-time PCR is to be performed on biopsies and not on swabs. These results show that panbacterial real-time PCR is a promising and quick method of determining the total bacterial load in chronic wounds, and suggest that it might be an important biomarker for the prognosis of chronic wounds under treatment.
Literature
1.
go back to reference Gjodsbol K, Christensen JJ, Karlsmark T, Jorgensen B, Klein BM, Krogfelt KA (2006) Multiple bacterial species reside in chronic wounds: a longitudinal study. Int Wound J 3:225–231PubMedCrossRef Gjodsbol K, Christensen JJ, Karlsmark T, Jorgensen B, Klein BM, Krogfelt KA (2006) Multiple bacterial species reside in chronic wounds: a longitudinal study. Int Wound J 3:225–231PubMedCrossRef
2.
go back to reference Frank DN, Wysocki A, Specht-Glick DD, Rooney A, Feldman RA, St. Amand AL, Pace NR, Trent JD (2009) Microbial diversity in chronic open wounds. Wound Repair Regen 17:163–172PubMedCrossRef Frank DN, Wysocki A, Specht-Glick DD, Rooney A, Feldman RA, St. Amand AL, Pace NR, Trent JD (2009) Microbial diversity in chronic open wounds. Wound Repair Regen 17:163–172PubMedCrossRef
3.
go back to reference Brook I, Frazier EH (1998) Aerobic and anaerobic microbiology of chronic venous ulcers. Int J Dermatol 37:426–428PubMedCrossRef Brook I, Frazier EH (1998) Aerobic and anaerobic microbiology of chronic venous ulcers. Int J Dermatol 37:426–428PubMedCrossRef
4.
go back to reference Howell-Jones RS, Wilson MJ, Hill KE, Howard AJ, Price PE, Thomas DW (2005) A review of the microbiology, antibiotic usage and resistance in chronic skin wounds. J Antimicrob Chemother 55:143–149PubMedCrossRef Howell-Jones RS, Wilson MJ, Hill KE, Howard AJ, Price PE, Thomas DW (2005) A review of the microbiology, antibiotic usage and resistance in chronic skin wounds. J Antimicrob Chemother 55:143–149PubMedCrossRef
5.
go back to reference Bowler PG, Duerden BI, Armstrong DG (2001) Wound microbiology and associated approaches to wound management. Clin Microbiol Rev 14:244–269PubMedCrossRef Bowler PG, Duerden BI, Armstrong DG (2001) Wound microbiology and associated approaches to wound management. Clin Microbiol Rev 14:244–269PubMedCrossRef
6.
go back to reference Bowler PG, Davies BJ (1999) The microbiology of infected and noninfected leg ulcers. Int J Dermatol 38:573–578PubMedCrossRef Bowler PG, Davies BJ (1999) The microbiology of infected and noninfected leg ulcers. Int J Dermatol 38:573–578PubMedCrossRef
7.
go back to reference Madsen SM, Westh H, Danielsen L, Rosdahl VT (1996) Bacterial colonisation and healing of venous leg ulcers. APMIS 104:895–899PubMedCrossRef Madsen SM, Westh H, Danielsen L, Rosdahl VT (1996) Bacterial colonisation and healing of venous leg ulcers. APMIS 104:895–899PubMedCrossRef
8.
go back to reference Wolcott RD, Dowd SE (2008) A rapid molecular method for characterising bacterial bioburden in chronic wounds. J Wound Care 17:513–516PubMed Wolcott RD, Dowd SE (2008) A rapid molecular method for characterising bacterial bioburden in chronic wounds. J Wound Care 17:513–516PubMed
9.
go back to reference Thomsen TR, Aasholm MS, Rudkjøbing VB, Saunders AM, Bjarnsholt T, Givskov M, Kirketerp-Møller K, Nielsen PH (2010) The bacteriology of chronic venous leg ulcer examined by culture-independent molecular methods. Wound Repair Regen 18:38–49PubMedCrossRef Thomsen TR, Aasholm MS, Rudkjøbing VB, Saunders AM, Bjarnsholt T, Givskov M, Kirketerp-Møller K, Nielsen PH (2010) The bacteriology of chronic venous leg ulcer examined by culture-independent molecular methods. Wound Repair Regen 18:38–49PubMedCrossRef
10.
go back to reference Martin JM, Zenilman JM, Lazarus GS (2010) Molecular microbiology: new dimensions for cutaneous biology and wound healing. J Invest Dermatol 130:38–48PubMedCrossRef Martin JM, Zenilman JM, Lazarus GS (2010) Molecular microbiology: new dimensions for cutaneous biology and wound healing. J Invest Dermatol 130:38–48PubMedCrossRef
11.
go back to reference Hill KE, Davies CE, Wilson MJ, Stephens P, Harding KG, Thomas DW (2003) Molecular analysis of the microflora in chronic venous leg ulceration. J Med Microbiol 52:365–369PubMedCrossRef Hill KE, Davies CE, Wilson MJ, Stephens P, Harding KG, Thomas DW (2003) Molecular analysis of the microflora in chronic venous leg ulceration. J Med Microbiol 52:365–369PubMedCrossRef
12.
go back to reference Fang H, Hedin G (2003) Rapid screening and identification of methicillin-resistant Staphylococcus aureus from clinical samples by selective-broth and real-time PCR assay. J Clin Microbiol 41:2894–2899PubMedCrossRef Fang H, Hedin G (2003) Rapid screening and identification of methicillin-resistant Staphylococcus aureus from clinical samples by selective-broth and real-time PCR assay. J Clin Microbiol 41:2894–2899PubMedCrossRef
13.
go back to reference Boutaga K, van Winkelhoff AJ, Vandenbroucke-Grauls CMJE, Savelkoul PHM (2005) Periodontal pathogens: a quantitative comparison of anaerobic culture and real-time PCR. FEMS Immunol Med Microbiol 45:191–199PubMedCrossRef Boutaga K, van Winkelhoff AJ, Vandenbroucke-Grauls CMJE, Savelkoul PHM (2005) Periodontal pathogens: a quantitative comparison of anaerobic culture and real-time PCR. FEMS Immunol Med Microbiol 45:191–199PubMedCrossRef
14.
go back to reference Wilson KH, Blitchington RB, Greene RC (1990) Amplification of bacterial 16S ribosomal DNA with polymerase chain reaction. J Clin Microbiol 28:1942–1946PubMed Wilson KH, Blitchington RB, Greene RC (1990) Amplification of bacterial 16S ribosomal DNA with polymerase chain reaction. J Clin Microbiol 28:1942–1946PubMed
15.
go back to reference Davies CE, Hill KE, Wilson MJ, Stephens P, Hill CM, Harding KG, Thomas DW (2004) Use of 16S ribosomal DNA PCR and denaturing gradient gel electrophoresis for analysis of the microfloras of healing and nonhealing chronic venous leg ulcers. J Clin Microbiol 42:3549–3557PubMedCrossRef Davies CE, Hill KE, Wilson MJ, Stephens P, Hill CM, Harding KG, Thomas DW (2004) Use of 16S ribosomal DNA PCR and denaturing gradient gel electrophoresis for analysis of the microfloras of healing and nonhealing chronic venous leg ulcers. J Clin Microbiol 42:3549–3557PubMedCrossRef
16.
go back to reference Doyle RJ, Rosenberg M (eds) (1990) Microbial cell surface hydrophobicity. ASM, Washington DC Doyle RJ, Rosenberg M (eds) (1990) Microbial cell surface hydrophobicity. ASM, Washington DC
17.
go back to reference Ljungh A, Wadström T (1995) Growth conditions influence expression of cell surface hydrophobicity of staphylococci and other wound infection pathogens. Microbiol Immunol 39:753–757PubMed Ljungh A, Wadström T (1995) Growth conditions influence expression of cell surface hydrophobicity of staphylococci and other wound infection pathogens. Microbiol Immunol 39:753–757PubMed
18.
go back to reference Ljungh A, Yanagisawa N, Wadström T (2006) Using the principle of hydrophobic interaction to bind and remove wound bacteria. J Wound Care 15:175–180PubMed Ljungh A, Yanagisawa N, Wadström T (2006) Using the principle of hydrophobic interaction to bind and remove wound bacteria. J Wound Care 15:175–180PubMed
19.
go back to reference O’Meara S, Cullum N, Majid M, Sheldon T (2000) Systematic reviews of wound care management: (3) antimicrobial agents for chronic wounds; (4) diabetic foot ulceration. Health Technol Assess 4:1–237 O’Meara S, Cullum N, Majid M, Sheldon T (2000) Systematic reviews of wound care management: (3) antimicrobial agents for chronic wounds; (4) diabetic foot ulceration. Health Technol Assess 4:1–237
20.
go back to reference Wright JB, Lam K, Burrell RE (1998) Wound management in an era of increasing bacterial antibiotic resistance: a role for topical silver treatment. Am J Infect Control 26:572–577PubMedCrossRef Wright JB, Lam K, Burrell RE (1998) Wound management in an era of increasing bacterial antibiotic resistance: a role for topical silver treatment. Am J Infect Control 26:572–577PubMedCrossRef
21.
go back to reference Yang S, Lin S, Kelen GD, Quinn TC, Dick JD, Gaydos CA, Rothman RE (2002) Quantitative multiprobe PCR assay for simultaneous detection and identification to species level of bacterial pathogens. J Clin Microbiol 40:3449–3454PubMedCrossRef Yang S, Lin S, Kelen GD, Quinn TC, Dick JD, Gaydos CA, Rothman RE (2002) Quantitative multiprobe PCR assay for simultaneous detection and identification to species level of bacterial pathogens. J Clin Microbiol 40:3449–3454PubMedCrossRef
22.
go back to reference Moore K, Huddleston E, Stacey MC, Harding KG (2007) Venous leg ulcer—the search for a prognostic indicator. Int Wound J 4:163–172PubMedCrossRef Moore K, Huddleston E, Stacey MC, Harding KG (2007) Venous leg ulcer—the search for a prognostic indicator. Int Wound J 4:163–172PubMedCrossRef
23.
go back to reference Donovan S (1998) Wound infection and wound swabbing. Prof Nurse 13:757–759PubMed Donovan S (1998) Wound infection and wound swabbing. Prof Nurse 13:757–759PubMed
24.
go back to reference Rantakokko-Jalava K, Nikkari S, Jalava J, Eerola E, Skurnik M, Meurman O, Ruuskanen O, Alanen A, Kotilainen E, Toivanen P, Kotilainen P (2000) Direct amplification of rRNA genes in diagnosis of bacterial infections. J Clin Microbiol 38:32–39PubMed Rantakokko-Jalava K, Nikkari S, Jalava J, Eerola E, Skurnik M, Meurman O, Ruuskanen O, Alanen A, Kotilainen E, Toivanen P, Kotilainen P (2000) Direct amplification of rRNA genes in diagnosis of bacterial infections. J Clin Microbiol 38:32–39PubMed
25.
go back to reference Fenollar F, Roux V, Stein A, Drancourt M, Raoult D (2006) Analysis of 525 samples to determine the usefulness of PCR amplification and sequencing of the 16S rRNA gene for diagnosis of bone and joint infections. J Clin Microbiol 44:1018–1028PubMedCrossRef Fenollar F, Roux V, Stein A, Drancourt M, Raoult D (2006) Analysis of 525 samples to determine the usefulness of PCR amplification and sequencing of the 16S rRNA gene for diagnosis of bone and joint infections. J Clin Microbiol 44:1018–1028PubMedCrossRef
26.
go back to reference Dowd SE, Delton Hanson J, Rees E, Wolcott RD, Zischau AM, Sun Y, White J, Smith DM, Kennedy J, Jones CE (2011) Survey of fungi and yeast in polymicrobial infections in chronic wounds. J Wound Care 20:40–47PubMed Dowd SE, Delton Hanson J, Rees E, Wolcott RD, Zischau AM, Sun Y, White J, Smith DM, Kennedy J, Jones CE (2011) Survey of fungi and yeast in polymicrobial infections in chronic wounds. J Wound Care 20:40–47PubMed
Metadata
Title
Panbacterial real-time PCR to evaluate bacterial burden in chronic wounds treated with Cutimed™ Sorbact™
Authors
V. Gentili
S. Gianesini
P. G. Balboni
E. Menegatti
A. Rotola
M. Zuolo
E. Caselli
P. Zamboni
D. Di Luca
Publication date
01-07-2012
Publisher
Springer-Verlag
Published in
European Journal of Clinical Microbiology & Infectious Diseases / Issue 7/2012
Print ISSN: 0934-9723
Electronic ISSN: 1435-4373
DOI
https://doi.org/10.1007/s10096-011-1473-x

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