Transfusion Medicine Reviews
Volume 21, Issue 3 , Pages 237-254 , July 2007

Real-Time Polymerase Chain Reaction in Transfusion Medicine: Applications for Detection of Bacterial Contamination in Blood Products

  • Jens Dreier

      Affiliations

    • Corresponding Author InformationAddress reprint requests to Jens Dreier, Institut für Laboratoriums und Transfusionsmedizin, Herz und Diabeteszentrum Nordrhein-Westfalen, Universitätsklinik der Ruhr-Universität Bochum, Georgstrasse 11, D-32545 Bad Oeynhausen, Germany.
  • ,
  • Melanie Störmer
  • ,
  • Knut Kleesiek

References 

  1. Brecher ME, Hay SN. Bacterial contamination of blood components. Clin Microbiol Rev. 2005;18:195–204
  2. Wagner SJ, Friedman LI, Dodd RY. Transfusion-associated bacterial sepsis. Clin Microbiol Rev. 1994;7:290–302
  3. De Korte D, Marcelis JH, Soeterboek AM. Determination of the degree of bacterial contamination of whole-blood collections using an automated microbe-detection system. Transfusion. 2001;41:815–818
  4. Kuehnert MJ, Roth VR, Haley NR, et al. Transfusion transmitted bacterial infection in the United States, 1998 through 2000. Transfusion. 2001;41:1493–1499
  5. Wagner SJ. Transfusion-transmitted bacterial infection: Risk, sources and interventions. Vox Sang. 2004;86:157–163
  6. Mohr H, Lambrecht B, Bayer A, et al. Sterility testing of platelet concentrates prepared from deliberately infected blood donations. Transfusion. 2006;46:486–491
  7. Boekhorst PA, Beckers EA, Vos MC, et al. Clinical significance of bacteriologic screening in platelet concentrates. Transfusion. 2005;45:514–519
  8. Walther-Wenke G, Doerner R, Montag T, et al. Bacterial contamination of platelet concentrates prepared by different methods: Results of standardized sterility testing in Germany. Vox Sang. 2006;90:177–182
  9. Brecher ME, Holland PV, Pineda AA, et al. Growth of bacteria in inoculated platelets: Implications for bacteria detection and the extension of platelet storage. Transfusion. 2000;40:1308–1312
  10. Dreier J, Störmer M, Kleesiek K. Two novel real-time reverse transcriptase PCR assays for rapid detection of bacterial contamination in platelet concentrates. J Clin Microbiol. 2004;42:4759–4764
  11. Dunne WM, Case LK, Isgriggs L, et al. In-house validation of the BACTEC 9240 blood culture system for detection of bacterial contamination in platelet concentrates. Transfusion. 2005;45:1138–1142
  12. Holme S, McAlister MB, Ortolano GA, et al. Enhancement of a culture-based bacterial detection system (eBDS) for platelet products based on measurement of oxygen consumption. Transfusion. 2005;45:984–993
  13. Jacobs MR, Bajaksouzian S, Windau A, et al. Evaluation of the Scansystem method for detection of bacterially contaminated platelets. Transfusion. 2005;45:265–269
  14. Mohammadi T, Pietersz R, Vandenbroucke-Grauls C, et al. Detection of bacteria in platelet concentrates: Comparison of broad-range real-time 16S rDNA polymerase chain reaction and automated culturing. Transfusion. 2005;45:731–736
  15. Montag T. Perspectives and limitations in the bacterial screening of platelet concentrates. J Lab Med. 2006;30:60–65
  16. Mohr H, Lambrecht B, Bayer A, et al. Basics of flow cytometry–based sterility testing of platelet concentrates. Transfusion. 2006;46:41–49
  17. Müller TH, Mohr H, Montag T. Methods for the detection of bacterial contamination in blood products. J Lab Med. 2006;30:74–90
  18. Störmer M, Kleesiek K, Dreier J. High-volume extraction of nucleic acids by magnetic bead technology allows ultrasensitive detection of bacteria in blood components. Clin Chem. 2007;[in press]
  19. Mohr H, Lambrecht B, Bayer A, et al. Elimination and multiplication of bacteria during preparation and storage of buffy coat–derived platelet concentrates. Transfusion. 2006;46:949–955
  20. Espy MJ, Uhl JR, Sloan LM, et al. Real-time PCR in clinical microbiology: Applications for routine laboratory testing. Clin Microbiol Rev. 2006;19:165–256
  21. Klein D. Quantification using real-time PCR technology: Applications and limitations. Trends Mol Med. 2002;8:257–260
  22. Kubista M, Andrade JM, Bengtsson M, et al. The real-time polymerase chain reaction. Mol Aspects Med. 2006;27:95–125
  23. Mothershed EA, Whitney AM. Nucleic acid–based methods for the detection of bacterial pathogens: Present and future considerations for the clinical laboratory. Clin Chim Acta. 2006;363:206–220
  24. Mackay IM. Real-time PCR in the microbiology laboratory. Clin Microbiol Infect. 2004;10:190–212
  25. Bustin SA. Real-time PCR. Encyclopedia of diagnostic genomics and proteomics. 2005;11:17–25
  26. Costa JM, Ernault P, Olivi M, et al. Chimeric LNA/DNA probes as a detection system for real-time PCR. Clin Biochem. 2004;37:930–932
  27. Letertre C, Perelle S, Dilasser F, et al. Evaluation of the performance of LNA and MGB probes in 5'-nuclease PCR assays. Mol Cell Probes. 2003;17:307–311
  28. Watzinger F, Ebner K, Lion T. Detection and monitoring of virus infections by real-time PCR. Mol Aspects Med. 2006;27:245–298
  29. Sen K. Rapid identification of Yersinia enterocolitica in blood by the 5′ nuclease PCR assay. J Clin Microbiol. 2000;38:1953–1958
  30. Sen K, Asher DM. Multiplex PCR for detection of Enterobacteriaceae in blood. Transfusion. 2001;41:1356–1364
  31. Feng P, Keasler SP, Hill WE. Direct identification of Yersinia enterocolitica in blood by polymerase chain reaction amplification. Transfusion. 1992;32S:850–854
  32. Petershofen EK, Fislage R, Faber R, et al. Detection of nucleic acid sequences from bacterial species with molecular genetic methods. Transfus Sci. 2000;23:21–27
  33. Harris KA, Hartley JC. Development of broad-range 16S rDNA PCR for use in the routine diagnostic clinical microbiology service. J Med Microbiol. 2003;52:685–691
  34. Jordan JA, Durso MB. Real-time polymerase chain reaction for detecting bacterial DNA directly from blood of neonates being evaluated for sepsis. J Mol Diagn. 2005;7:575–581
  35. Peters RP, Mohammadi T, Vandenbroucke-Grauls CM, et al. Detection of bacterial DNA in blood samples from febrile patients: Underestimated infection or emerging contamination?. FEMS Immunol Med Microbiol. 2004;42:249–253
  36. Picard FJ, Ke D, Boudreau DK, et al. Use of tuf sequences for genus-specific PCR detection and phylogenetic analysis of 28 streptococcal species. J Clin Microbiol. 2004;42:3686–3695
  37. Drancourt M, Roux V, Fournier PE, et al. rpoB gene sequence-based identification of aerobic Gram-positive cocci of the genera Streptococcus, Enterococcus, Gemella, Abiotrophia, and Granulicatella. J Clin Microbiol. 2004;42:497–504
  38. Goh SH, Potter S, Wood JO, et al. HSP60 gene sequences as universal targets for microbial species identification: studies with coagulase-negative staphylococci. J Clin Microbiol. 1996;34:818–823
  39. Ludwig W, Strunk O, Klugbauer S, et al. Bacterial phylogeny based on comparative sequence analysis. Electrophoresis. 1998;19:554–568
  40. Weisburg WG, Barns SM, Pelletier DA, et al. 16S ribosomal DNA amplification for phylogenetic study. J Bacteriol. 1991;173:697–703
  41. Jensen MA, Webster JA, Straus N. Rapid identification of bacteria on the basis of polymerase chain reaction–amplified ribosomal DNA spacer polymorphisms. Appl Environ Microbiol. 1993;59:945–952
  42. Farelly V, Rainley FA, Stackebrandt E. Effect of genome size and rrn gene copy number on PCR amplification of 16S rRNA genes from a mixture of bacterial species. Appl Environ Microbiol. 1995;6-1:2798–2801
  43. Gurtler V, Stanisich VA. New approaches to typing and identification of bacteria using 16S-23S rDNA spacer region. Microbiology. 1996;142:3–116
  44. Nadkarni MA, Martin FE, Jacques NA, et al. Determination of bacterial load by real-time PCR using a broad-range (universal) probe and primers set. Microbiology. 2002;148:257–266
  45. Watson JD, Hopkins NH, Roberts JW, et al. In: Molecular biology of the gene. vol 1:Menlo Park (CA): The Benjamin/Cummings Publishing Company; 1987;p. 466
  46. Keer JT, Birch L. Molecular methods for the assessment of bacterial viability. J Microbiol Methods. 2003;53:175–183
  47. Pedersen S, Reeh S. Functional mRNA half lives in E. coli. Mol Gen Genet. 1978;166:329–336
  48. Bernstein JA, Khodursky AB, Lin PH, et al. Global analysis of mRNA decay and abundance in Escherichia coli at single-gene resolution using two-color fluorescent DNA microarrays. Proc Natl Acad Sci U S A. 2002;99:9697–9702
  49. Sheridan GEC, Masters CL, Shallcross JA, et al. Detection of mRNA by reverse transcription–PCR as an indicator of viability in Escherichia coli cells. Appl Environ Microbiol. 1998;64:1313–1318
  50. Rothman RE, Majmudar MD, Kelen GD, et al. Detection of bacteremia in emergency department patients at risk for infective endocarditis using universal 16S rRNA primers in a decontaminated polymerase chain reaction assay. J Infect Dis. 2002;186:1677–1681
  51. Peters RP, van Agtmael MA, Danner SA, et al. New developments in the diagnosis of bloodstream infections. Lancet Infect Dis. 2004;4:751–760
  52. Rovery C, Greub G, Lepidi H, et al. PCR detection of bacteria on cardiac valves of patients with treated bacterial endocarditis. J Clin Microbiol. 2005;43:163–167
  53. Goldenberger D, Kunzli A, Vogt P, et al. Molecular diagnosis of bacterial endocarditis by broad-range PCR amplification and direct sequencing. J Clin Microbiol. 1997;35:2733–2739
  54. Breitkopf C, Hammel D, Scheld HH, et al. Impact of a molecular approach to improve the microbiological diagnosis of infective heart valve endocarditis. Circulation. 2005;111:1415–1421
  55. Schuurman T, de Boer RF, Kooistra-Smid AM, et al. Prospective study of use of PCR amplification and sequencing of 16S ribosomal DNA from cerebrospinal fluid for diagnosis of bacterial meningitis in a clinical setting. J Clin Microbiol. 2004;42:734–740
  56. Xu J, Moore JE, Millar BC, et al. Employment of broad range 16S rDNA PCR and sequencing in the detection of aetiological agents of meningitis. New Microbiol. 2005;10 28:135–143
  57. Hoorfar J, Wolffs P, Radstrom P. Diagnostic PCR: validation and sample preparation are two sides of the same coin. APMIS. 2004;112:808–814
  58. Apfalter P, Reischl U, Hammerschlag MR. In-house nucleic acid amplification assays in research: How much quality control is needed before one can rely upon the results?. J Clin Microbiol. 2005;43:5835–5841
  59. Anonymous . ICH topic Q2A: Validation of analytical methods: Definitions and terminology. CPMP/ICH/381/95. http://www.eudra.org/eudraportal/
  60. Anonymous . ICH topic Q2B: Validation of analytical methods: Methodology. CPMP/ICH/281/95. http://www.eudra.org/eudraportal/
  61. Anonymous. European Pharmacopeia: PCR monograph
  62. Saldanha J. Standardization: A progress report. Biologicals. 1999;27:285–289
  63. Saldanha J. Validation and standardisation of nucleic acid amplification technology (NAT) assays for the detection of viral contamination of blood and blood products. J Clin Virol. 2001;20:7–13
  64. Montag T. Concepts for bacterial safety of blood components in Germany. Transfus Apher Sci. 2001;24:251–252
  65. Schmidt M, Weis C, Heck J, et al. Optimized Scansystem platelet kit for bacterial detection within 24 h after spiking. Vox Sang. 2005;89:135–139
  66. Störmer M, Cassens U, Kleesiek K, et al. Detection of bacteria in platelet concentrates prepared from spiked single donations using cultural and molecular genetic methods. Transfus Med. 2007;17:61–70
  67. Amikam D, Glaser G, Razin S. Mycoplasmas (Mollicutes) have a low number of rRNA genes. J Bacteriol. 1984;158:376–378
  68. Mohammadi T, Reesink HW, Vandenbroucke-Grauls CMJE, et al. Removal of contaminating DNA from commercial nucleic acid extraction kit reagents. J Microbiol Methods. 2005;61:285–288
  69. Wellinghausen N, Frost C, Marre R. Detection of legionellae in hospital water samples by quantitative real-time LightCycler PCR. Appl Environ Microbiol. 2001;67:3985–3993
  70. Dreier J, Störmer M, Kleesiek K. Use of bacteriophage MS2 as internal control in viral RT-PCR assays. J Clin Microbiol. 2005;43:4551–4557
  71. Hennig H, Luhm J, Hartwig D, et al. A novel RT-PCR for reliable and rapid HCV RNA screening of blood donations. Transfusion. 2001;41:1100–1106
  72. Rider J, Newton A. Electrochemiluminescent detection of bacteria in blood components. Transfus Med. 2002;12:115–123
  73. Chaney R, Rider J, Pamphilon D. Direct detection of bacteria in cellular blood products using bacterial ribosomal RNA–directed probes coupled to electrochemiluminescence. Transfus Med. 1999;9:177–188
  74. Brecher ME, Hogan JJ, Boothe G. Platelet bacterial contamination and the use of a chemiluminescence-linked universal bacterial ribosomal RNA gene probe. Transfusion. 1994;34:750–755
  75. Brecher ME, Hogan JJ, Boothe G. The use of a chemiluminescence-linked universal bacterial ribosomal RNA gene probe and blood gas analysis for the rapid detection of bacterial contamination in white cell reduced and nonreduced platelets. Transfusion. 1993;33:450–457
  76. Mohammadi T, Reesink HW, Vandenbroucke-Grauls CM, et al. Optimization of real-time PCR assay for rapid and sensitive detection of eubacterial 16S ribosomal DNA in platelet concentrates. J Clin Microbiol. 2003;41:4796–4798
  77. Klaschik S, Lehmann LE, Raadts A, et al. Detection and differentiation of in vitro–spiked bacteria by real-time PCR and melting-curve analysis. J Clin Microbiol. 2004;42:512–517
  78. Kunishima S, Inoue C, Kamiya T, et al. Presence of Propionibacterium acnes in blood components. Transfusion. 2001;41:1126–1129
  79. Karahan ZC, Mumcuoglu I, Guriz H, et al. PCR evaluation of false-positive signals from two automated blood-culture systems. J Med Microbiol. 2006;55:53–57
  80. Turenne CY, Witwicki E, Hoban DJ, et al. Rapid identification of bacteria from positive blood cultures by fluorescence-based PCR–single-strand conformation polymorphism analysis of the 16S rRNA gene. J Clin Microbiol. 2000;38:513–520
  81. Mohammadi T, Pietersz R, Scholtalbers L, et al. Optimal sampling time after preparation of platelet concentrates for detection of bacterial contamination by quantitative real-time polymerase chain reaction. Vox Sang. 2005;89:208–214
  82. Blajchman MA, Goldman M, Baeza F. Improving the bacteriological safety of platelet transfusion. Transfus Med Rev. 2004;18:11–24
  83. Vasconcelos E, Seghatchian J. Bacterial contamination in blood components and preventative strategies: An overview. Transfus Apher Sci. 2004;31:155–163
  84. Muder RR, Yee YC, Rihs J, et al. Staphylococcus epidermidis bacteremia from transfusion of contaminated platelets: application of bacterial DNA analysis. Transfusion. 1992;32:771–774
  85. Heal JM, Singal S, Sardisco E, et al. Bacterial proliferation in platelet concentrates. Transfusion. 1986;26:388–390
  86. Ribault S, Harper K, Grave L, et al. Rapid screening method for detection of bacteria in platelet concentrates. J Clin Microbiol. 2004;42:1903–1908
  87. Macauley A, Chandrasekar A, Geddis G, et al. Operational feasibility of routine bacterial monitoring of platelets. Transfus Med. 2003;13:189–195
  88. Ortolano GA, Freundlich LF, Holme S, et al. Detection of bacteria in WBC-reduced PLT concentrates using percent oxygen as a marker for bacteria growth. Transfusion. 2003;43:1276–1283
  89. Fournier-Wirth C, Deschaseaux M, Defer C, et al. Evaluation of the enhanced bacterial detection system for screening of contaminated platelets. Transfusion. 2006;46:220–224
  90. Rantakokko-Javala K, Javala J. Optimal DNA isolation method for detection of bacteria in clinical specimens by broad-range PCR. J Clin Microbiol. 2002;40:4211–4217
  91. Beuselinck K, Ranst M, Eldere J. Automated extraction of viral-pathogen RNA and DNA for high-throughput quantitative real-time PCR. J Clin Microbiol. 2005;43:5541–5546
  92. Hourfar MK, Schmidt M, Seifried E, et al. Evaluation of an automated high-volume extraction method for viral nucleic acids in comparison to a manual procedure with preceding enrichment. Vox Sang. 2005;89:71–76
  93. Pichl L, Heitmann A, Herzog P, et al. Magnetic bead technology in viral RNA and DNA extraction from plasma minipools. Transfusion. 2005;45:1106–1110
  94. Kleines M, Schellenberg K, Ritter K. Efficient extraction of viral DNA and viral RNA by the Chemagic viral DNA/RNA Kit allows sensitive detection of cytomegalovirus, hepatitis B virus, and hepatitis G virus by PCR. J Clin Microbiol. 2003;41:5273–5276
  95. Millar BC, Xu J, Moore JE. Risk assessment models and contamination management: implications for broad-range ribosomal DNA PCR as a diagnostic tool in medical bacteriology. J Clin Microbiol. 2002;40:1575–1580
  96. Longo MC, Berninger MS, Hartley JL. Use of uracil DNA glycosylase to control carry-over contamination in polymerase chain reactions. Gene. 1990;93:125–128
  97. Böttger EC. Frequent contamination of Taq polymerase with DNA. Clin Chem. 1990;36:1258
  98. Rand VH, Houck H. Taq polymerase contains bacterial DNA of unknown origin. Mol Cell Probes. 1990;4:445–450
  99. Corless CE, Guiver M, Borrow R, et al. Contamination and sensitivity issues with a real-time universal 16S rRNA PCR. J Clin Microbiol. 2000;38:1747–1752
  100. Carroll NM, Adamson P, Okhravi N. Elimination of bacterial DNA from Taq DNA polymerases by restriction endonuclease digestion. J Clin Microbiol. 1999;37:3402–3404
  101. Meier A, Persing DH, Finken M, Bottger EC. Elimination of contaminating DNA within polymerase chain reactions reagents: Implications for a general approach to detection of uncultured pathogens. J Clin Microbiol. 1993;31:646–652
  102. Jinno Y, Yoshiura K, Niikawa N. Use of psoralen as extinguisher of contaminated DNA in PCR. Nucleic Acids Res. 1990;18:6739
  103. Barrett BB, Andersen JW, Anderson KC. Strategies for the avoidance of bacterial contamination of blood components. Transfusion. 1993;33:228–233
  104. Pink JM, MacCallum S, Ribeiro A, et al. Platelet transfusion–related sepsis. Aust N Z J Med. 1993;23:717

PII: S0887-7963(07)00021-1

doi: 10.1016/j.tmrv.2007.03.006

Transfusion Medicine Reviews
Volume 21, Issue 3 , Pages 237-254 , July 2007