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Published in: Critical Care 4/2013

Open Access 01-08-2013 | Research

Storage of bronchoalveolar lavage fluid and accuracy of microbiologic diagnostics in the ICU: a prospective observational study

Authors: Nikolaus Kneidinger, Joanna Warszawska, Peter Schenk, Valentin Fuhrmann, Andja Bojic, Alexander Hirschl, Harald Herkner, Christian Madl, Athanasios Makristathis

Published in: Critical Care | Issue 4/2013

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Abstract

Introduction

Early initiation of appropriate antimicrobial treatment is a cornerstone in managing pneumonia. Because microbiologic processing may not be available around the clock, optimal storage of specimens is essential for accurate microbiologic identification of pathogenetic bacteria. The aim of our study was to determine the accuracy of two commonly used storage approaches for delayed processing of bronchoalveolar lavage in critically ill patients with suspected pneumonia.

Methods

This study included 132 patients with clinically suspected pneumonia at two medical intensive care units of a tertiary care hospital. Bronchoalveolar lavage samples were obtained and divided into three aliquots: one was used for immediate culture, and two, for delayed culture (DC) after storage for 24 hours at 4°C (DC4) and -80°C (DC-80), respectively.

Results

Of 259 bronchoalveolar lavage samples, 84 (32.4%) were positive after immediate culture with 115 relevant culture counts (≥104 colony-forming units/ml). Reduced (<104 colony-forming units/ml) or no growth of four and 57 of these isolates was observed in DC4 and DC-80, respectively. The difference between mean bias of immediate culture and DC4 (-0.035; limits of agreement, -0.977 to 0.906) and immediate culture and DC-80 (-1.832; limits of agreement, -4.914 to 1.267) was -1.788 ± 1.682 (P < 0.0001). Sensitivity and negative predictive value were 96.5% and 97.8% for DC4 and 50.4% and 75.4% for DC-80, respectively; the differences were statistically significant (P < 0.0001).

Conclusions

Bronchoalveolar lavage samples can be processed for culture when stored up to 24 hours at 4°C without loss of diagnostic accuracy. Delayed culturing after storage at -80°C may not be reliable, in particular with regard to Gram-negative bacteria.
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Literature
1.
go back to reference Richards MJ, Edwards JR, Culver DH, Gaynes RP: Nosocomial infections in medical intensive care units in the United States: National Nosocomial Infections Surveillance System. Crit Care Med. 1999, 27: 887-892. 10.1097/00003246-199905000-00020.PubMedCrossRef Richards MJ, Edwards JR, Culver DH, Gaynes RP: Nosocomial infections in medical intensive care units in the United States: National Nosocomial Infections Surveillance System. Crit Care Med. 1999, 27: 887-892. 10.1097/00003246-199905000-00020.PubMedCrossRef
2.
go back to reference American Thoracic Society, Infectious Diseases Society of America: Guidelines for the management of adults with hospital-acquired, ventilator-associated, and healthcare-associated pneumonia. Am J Respir Crit Care Med. 2005, 171: 388-416.CrossRef American Thoracic Society, Infectious Diseases Society of America: Guidelines for the management of adults with hospital-acquired, ventilator-associated, and healthcare-associated pneumonia. Am J Respir Crit Care Med. 2005, 171: 388-416.CrossRef
3.
go back to reference Safdar N, Dezfulian C, Collard HR, Saint S: Clinical and economic consequences of ventilator-associated pneumonia: a systematic review. Crit Care Med. 2005, 33: 2184-2193. 10.1097/01.CCM.0000181731.53912.D9.PubMedCrossRef Safdar N, Dezfulian C, Collard HR, Saint S: Clinical and economic consequences of ventilator-associated pneumonia: a systematic review. Crit Care Med. 2005, 33: 2184-2193. 10.1097/01.CCM.0000181731.53912.D9.PubMedCrossRef
4.
go back to reference Bercault N, Boulain T: Mortality rate attributable to ventilator-associated nosocomial pneumonia in an adult intensive care unit: a prospective case-control study. Crit Care Med. 2001, 29: 2303-2309. 10.1097/00003246-200112000-00012.PubMedCrossRef Bercault N, Boulain T: Mortality rate attributable to ventilator-associated nosocomial pneumonia in an adult intensive care unit: a prospective case-control study. Crit Care Med. 2001, 29: 2303-2309. 10.1097/00003246-200112000-00012.PubMedCrossRef
5.
go back to reference Iregui M, Ward S, Sherman G, Fraser VJ, Kollef MH: Clinical importance of delays in the initiation of appropriate antibiotic treatment for ventilator-associated pneumonia. Chest. 2002, 122: 262-268. 10.1378/chest.122.1.262.PubMedCrossRef Iregui M, Ward S, Sherman G, Fraser VJ, Kollef MH: Clinical importance of delays in the initiation of appropriate antibiotic treatment for ventilator-associated pneumonia. Chest. 2002, 122: 262-268. 10.1378/chest.122.1.262.PubMedCrossRef
6.
go back to reference Luna CM, Aruj P, Niederman MS, Garzón J, Violi D, Prignoni A, Ríos F, Baquero S, Gando S, Grupo Argentino de Estudio de la Neumonía Asociada al Respirador group: Appropriateness and delay to initiate therapy in ventilator-associated pneumonia. Eur Respir J. 2006, 27: 158-164. 10.1183/09031936.06.00049105.PubMedCrossRef Luna CM, Aruj P, Niederman MS, Garzón J, Violi D, Prignoni A, Ríos F, Baquero S, Gando S, Grupo Argentino de Estudio de la Neumonía Asociada al Respirador group: Appropriateness and delay to initiate therapy in ventilator-associated pneumonia. Eur Respir J. 2006, 27: 158-164. 10.1183/09031936.06.00049105.PubMedCrossRef
7.
go back to reference Fagon JY, Chastre J, Rouby JJ: Is bronchoalveolar lavage with quantitative cultures a useful tool for diagnosing ventilator-associated pneumonia?. Crit Care. 2007, 11: 123-10.1186/cc5724.PubMedPubMedCentralCrossRef Fagon JY, Chastre J, Rouby JJ: Is bronchoalveolar lavage with quantitative cultures a useful tool for diagnosing ventilator-associated pneumonia?. Crit Care. 2007, 11: 123-10.1186/cc5724.PubMedPubMedCentralCrossRef
8.
go back to reference Canadian Critical Care Trials Group: A randomized trial of diagnostic techniques for ventilator-associated pneumonia. N Engl J Med. 2006, 355: 2619-2630.CrossRef Canadian Critical Care Trials Group: A randomized trial of diagnostic techniques for ventilator-associated pneumonia. N Engl J Med. 2006, 355: 2619-2630.CrossRef
9.
go back to reference Chastre J, Fagon JY: Ventilator-associated pneumonia. Am J Respir Crit Care Med. 2002, 165: 867-903. 10.1164/ajrccm.165.7.2105078.PubMedCrossRef Chastre J, Fagon JY: Ventilator-associated pneumonia. Am J Respir Crit Care Med. 2002, 165: 867-903. 10.1164/ajrccm.165.7.2105078.PubMedCrossRef
10.
go back to reference Prats E, Dorca J, Pujol M, Garcia L, Barreiro B, Verdaguer R, Gudiol F, Manresa F: Effects of antibiotics on protected specimen brush sampling in ventilator-associated pneumonia. Eur Respir J. 2002, 19: 944-951. 10.1183/09031936.02.00239302.PubMedCrossRef Prats E, Dorca J, Pujol M, Garcia L, Barreiro B, Verdaguer R, Gudiol F, Manresa F: Effects of antibiotics on protected specimen brush sampling in ventilator-associated pneumonia. Eur Respir J. 2002, 19: 944-951. 10.1183/09031936.02.00239302.PubMedCrossRef
11.
go back to reference Middleton RM, Huff W, Brickey DA, Kirkpatrick MB: Comparison of quantitative cultures to semiquantitative loop cultures of bronchoscopic protected specimen brush samples. Chest. 1996, 109: 1204-1209. 10.1378/chest.109.5.1204.PubMedCrossRef Middleton RM, Huff W, Brickey DA, Kirkpatrick MB: Comparison of quantitative cultures to semiquantitative loop cultures of bronchoscopic protected specimen brush samples. Chest. 1996, 109: 1204-1209. 10.1378/chest.109.5.1204.PubMedCrossRef
12.
go back to reference Georges H, Santre C, Leroy O, Roussel-Delvallez M, Caillaux M, Beuscart C, Guery B, Vandenbussche C, Beaucaire G: Reliability of quantitative cultures of protected specimen brush after freezing. Am J Respir Crit Care Med. 1996, 153: 855-857. 10.1164/ajrccm.153.2.8564144.PubMedCrossRef Georges H, Santre C, Leroy O, Roussel-Delvallez M, Caillaux M, Beuscart C, Guery B, Vandenbussche C, Beaucaire G: Reliability of quantitative cultures of protected specimen brush after freezing. Am J Respir Crit Care Med. 1996, 153: 855-857. 10.1164/ajrccm.153.2.8564144.PubMedCrossRef
13.
go back to reference de Lassence A, Joly-Guillou ML, Martin-Lefevre L, Le Mière E, Lasry S, Morelot C, Coste F, Dreyfuss D: Accuracy of delayed cultures of plugged telescoping catheter samples for diagnosing bacterial pneumonia. Crit Care Med. 2001, 29: 1311-1317. 10.1097/00003246-200107000-00003.PubMedCrossRef de Lassence A, Joly-Guillou ML, Martin-Lefevre L, Le Mière E, Lasry S, Morelot C, Coste F, Dreyfuss D: Accuracy of delayed cultures of plugged telescoping catheter samples for diagnosing bacterial pneumonia. Crit Care Med. 2001, 29: 1311-1317. 10.1097/00003246-200107000-00003.PubMedCrossRef
14.
go back to reference de Lassence A, Joly-Guillou ML, Salah A, Martin-Lefevre L, Hidri N, Ricard JD, Bouvry D, de Castro N, Dreyfuss D: Accuracy of delayed (24 hours) processing of bronchoalveolar lavage for diagnosing bacterial pneumonia. Crit Care Med. 2004, 32: 680-685. 10.1097/01.CCM.0000114813.85853.EA.PubMedCrossRef de Lassence A, Joly-Guillou ML, Salah A, Martin-Lefevre L, Hidri N, Ricard JD, Bouvry D, de Castro N, Dreyfuss D: Accuracy of delayed (24 hours) processing of bronchoalveolar lavage for diagnosing bacterial pneumonia. Crit Care Med. 2004, 32: 680-685. 10.1097/01.CCM.0000114813.85853.EA.PubMedCrossRef
15.
go back to reference Andrews CP, Coalson JJ, Smith JD, Johanson WG: Diagnosis of nosocomial bacterial pneumonia in acute, diffuse lung injury. Chest. 1981, 80: 254-258. 10.1378/chest.80.3.254.PubMedCrossRef Andrews CP, Coalson JJ, Smith JD, Johanson WG: Diagnosis of nosocomial bacterial pneumonia in acute, diffuse lung injury. Chest. 1981, 80: 254-258. 10.1378/chest.80.3.254.PubMedCrossRef
16.
go back to reference Le Gall JR, Lemeshow S, Saulnier F: A new Simplified Acute Physiology Score (SAPS II) based on a European/North American multicenter study. JAMA. 1993, 270: 2957-2963. 10.1001/jama.1993.03510240069035.PubMedCrossRef Le Gall JR, Lemeshow S, Saulnier F: A new Simplified Acute Physiology Score (SAPS II) based on a European/North American multicenter study. JAMA. 1993, 270: 2957-2963. 10.1001/jama.1993.03510240069035.PubMedCrossRef
17.
go back to reference Chastre J, Fagon JY, Soler P, Bornet M, Domart Y, Trouillet JL, Gibert C, Hance AJ: Diagnosis of nosocomial bacterial pneumonia in intubated patients undergoing ventilation: comparison of the usefulness of bronchoalveolar lavage and the protected specimen brush. Am J Med. 1988, 85: 499-506. 10.1016/S0002-9343(88)80085-8.PubMedCrossRef Chastre J, Fagon JY, Soler P, Bornet M, Domart Y, Trouillet JL, Gibert C, Hance AJ: Diagnosis of nosocomial bacterial pneumonia in intubated patients undergoing ventilation: comparison of the usefulness of bronchoalveolar lavage and the protected specimen brush. Am J Med. 1988, 85: 499-506. 10.1016/S0002-9343(88)80085-8.PubMedCrossRef
18.
go back to reference Bland JM, Altman DG: Statistical methods for assessing agreement between two methods of clinical measurement. Lancet. 1986, 1: 307-310.PubMedCrossRef Bland JM, Altman DG: Statistical methods for assessing agreement between two methods of clinical measurement. Lancet. 1986, 1: 307-310.PubMedCrossRef
19.
go back to reference Fagon JY, Chastre J, Wolff M, Gervais C, Parer-Aubas S, Stéphan F, Similowski T, Mercat A, Diehl JL, Sollet JP, Tenaillon A: A Invasive and noninvasive strategies for management of suspected ventilator-associated pneumonia: a randomized trial. Ann Intern Med. 2000, 132: 621-630.PubMedCrossRef Fagon JY, Chastre J, Wolff M, Gervais C, Parer-Aubas S, Stéphan F, Similowski T, Mercat A, Diehl JL, Sollet JP, Tenaillon A: A Invasive and noninvasive strategies for management of suspected ventilator-associated pneumonia: a randomized trial. Ann Intern Med. 2000, 132: 621-630.PubMedCrossRef
20.
go back to reference Sanchez-Nieto JM, Torres A, Garcia-Cordoba F, El-Ebiary M, Carrillo A, Ruiz J, Nuñez ML, Niederman M: Impact of invasive and noninvasive quantitative culture sampling on outcome of ventilator-associated pneumonia: a pilot study. Am J Respir Crit Care Med. 1998, 157: 371-376. 10.1164/ajrccm.157.2.97-02039.PubMedCrossRef Sanchez-Nieto JM, Torres A, Garcia-Cordoba F, El-Ebiary M, Carrillo A, Ruiz J, Nuñez ML, Niederman M: Impact of invasive and noninvasive quantitative culture sampling on outcome of ventilator-associated pneumonia: a pilot study. Am J Respir Crit Care Med. 1998, 157: 371-376. 10.1164/ajrccm.157.2.97-02039.PubMedCrossRef
21.
go back to reference Ruiz M, Torres A, Ewig S, Marcos MA, Alcón A, Lledó R, Asenjo MA, Maldonaldo A: Noninvasive versus invasive microbial investigation in ventilator-associated pneumonia: evaluation of outcome. Am J Respir Crit Care Med. 2000, 162: 119-125. 10.1164/ajrccm.162.1.9907090.PubMedCrossRef Ruiz M, Torres A, Ewig S, Marcos MA, Alcón A, Lledó R, Asenjo MA, Maldonaldo A: Noninvasive versus invasive microbial investigation in ventilator-associated pneumonia: evaluation of outcome. Am J Respir Crit Care Med. 2000, 162: 119-125. 10.1164/ajrccm.162.1.9907090.PubMedCrossRef
22.
go back to reference Sole Violan J, Fernandez JA, Benitez AB, Cardeñosa Cendrero JA, Rodríguez de Castro F: Impact of quantitative invasive diagnostic techniques in the management and outcome of mechanically ventilated patients with suspected pneumonia. Crit Care Med. 2000, 28: 2737-2741. 10.1097/00003246-200008000-00009.PubMedCrossRef Sole Violan J, Fernandez JA, Benitez AB, Cardeñosa Cendrero JA, Rodríguez de Castro F: Impact of quantitative invasive diagnostic techniques in the management and outcome of mechanically ventilated patients with suspected pneumonia. Crit Care Med. 2000, 28: 2737-2741. 10.1097/00003246-200008000-00009.PubMedCrossRef
23.
go back to reference Heyland DK, Cook DJ, Marshall J, Heule M, Guslits B, Lang J, Jaeschke R: The clinical utility of invasive diagnostic techniques in the setting of ventilator-associated pneumonia: Canadian Critical Care Trials Group. Chest. 1999, 115: 1076-1084. 10.1378/chest.115.4.1076.PubMedCrossRef Heyland DK, Cook DJ, Marshall J, Heule M, Guslits B, Lang J, Jaeschke R: The clinical utility of invasive diagnostic techniques in the setting of ventilator-associated pneumonia: Canadian Critical Care Trials Group. Chest. 1999, 115: 1076-1084. 10.1378/chest.115.4.1076.PubMedCrossRef
24.
go back to reference Fagon JY, Chastre J, Domart Y, Trouillet JL, Pierre J, Darne C, Gibert C: Nosocomial pneumonia in patients receiving continuous mechanical ventilation: prospective analysis of 52 episodes with use of a protected specimen brush and quantitative culture techniques. Am Rev Respir Dis. 1989, 139: 877-884. 10.1164/ajrccm/139.4.877.PubMedCrossRef Fagon JY, Chastre J, Domart Y, Trouillet JL, Pierre J, Darne C, Gibert C: Nosocomial pneumonia in patients receiving continuous mechanical ventilation: prospective analysis of 52 episodes with use of a protected specimen brush and quantitative culture techniques. Am Rev Respir Dis. 1989, 139: 877-884. 10.1164/ajrccm/139.4.877.PubMedCrossRef
25.
go back to reference Rello J, Jubert P, Valles J, Artigas A, Rué M, Niederman MS: Evaluation of outcome for intubated patients with pneumonia due to Pseudomonas aeruginosa. Clin Infect Dis. 1996, 23: 973-978. 10.1093/clinids/23.5.973.PubMedCrossRef Rello J, Jubert P, Valles J, Artigas A, Rué M, Niederman MS: Evaluation of outcome for intubated patients with pneumonia due to Pseudomonas aeruginosa. Clin Infect Dis. 1996, 23: 973-978. 10.1093/clinids/23.5.973.PubMedCrossRef
26.
go back to reference Kollef MH, Silver P, Murphy DM, Trovillion E: The effect of late-onset ventilator-associated pneumonia in determining patient mortality. Chest. 1995, 108: 1655-1662. 10.1378/chest.108.6.1655.PubMedCrossRef Kollef MH, Silver P, Murphy DM, Trovillion E: The effect of late-onset ventilator-associated pneumonia in determining patient mortality. Chest. 1995, 108: 1655-1662. 10.1378/chest.108.6.1655.PubMedCrossRef
27.
go back to reference Fagon JY, Chastre J, Hance AJ, Montravers P, Novara A, Gibert C: Nosocomial pneumonia in ventilated patients: a cohort study evaluating attributable mortality and hospital stay. Am J Med. 1993, 94: 281-288. 10.1016/0002-9343(93)90060-3.PubMedCrossRef Fagon JY, Chastre J, Hance AJ, Montravers P, Novara A, Gibert C: Nosocomial pneumonia in ventilated patients: a cohort study evaluating attributable mortality and hospital stay. Am J Med. 1993, 94: 281-288. 10.1016/0002-9343(93)90060-3.PubMedCrossRef
28.
go back to reference Arnold HM, Sawyer AM, Kollef MH: Use of adjunctive aerosolized antimicrobial therapy in the treatment of Pseudomonas aeruginosa and Acinetobacter baumannii ventilator-associated pneumonia. Respir Care. 2012, 1226-1233. 57 Arnold HM, Sawyer AM, Kollef MH: Use of adjunctive aerosolized antimicrobial therapy in the treatment of Pseudomonas aeruginosa and Acinetobacter baumannii ventilator-associated pneumonia. Respir Care. 2012, 1226-1233. 57
29.
go back to reference Peña C, Gómez-Zorrilla S, Oriol I, Tubau F, Dominguez MA, Pujol M, Ariza J: Impact of multidrug resistance on Pseudomonas aeruginosa ventilator-associated pneumonia outcome: predictors of early and crude mortality. Eur J Clin Microbiol Infect Dis. 2013, 32: 413-420. 10.1007/s10096-012-1758-8.PubMedCrossRef Peña C, Gómez-Zorrilla S, Oriol I, Tubau F, Dominguez MA, Pujol M, Ariza J: Impact of multidrug resistance on Pseudomonas aeruginosa ventilator-associated pneumonia outcome: predictors of early and crude mortality. Eur J Clin Microbiol Infect Dis. 2013, 32: 413-420. 10.1007/s10096-012-1758-8.PubMedCrossRef
30.
go back to reference Forceville X, Fiacre A, Faibis F, Lahilaire P, Demachy MC, Combes A: Reproducibility of protected specimen brush and bronchoalveolar lavage conserved at 4 degrees C for 48 hours. Intensive Care Med. 2002, 28: 857-863. 10.1007/s00134-002-1329-z.PubMedCrossRef Forceville X, Fiacre A, Faibis F, Lahilaire P, Demachy MC, Combes A: Reproducibility of protected specimen brush and bronchoalveolar lavage conserved at 4 degrees C for 48 hours. Intensive Care Med. 2002, 28: 857-863. 10.1007/s00134-002-1329-z.PubMedCrossRef
Metadata
Title
Storage of bronchoalveolar lavage fluid and accuracy of microbiologic diagnostics in the ICU: a prospective observational study
Authors
Nikolaus Kneidinger
Joanna Warszawska
Peter Schenk
Valentin Fuhrmann
Andja Bojic
Alexander Hirschl
Harald Herkner
Christian Madl
Athanasios Makristathis
Publication date
01-08-2013
Publisher
BioMed Central
Published in
Critical Care / Issue 4/2013
Electronic ISSN: 1364-8535
DOI
https://doi.org/10.1186/cc12814

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