The Pan Genera Detection Immunoassay: a Novel Point-of-Issue Method for Detection of Bacterial Contamination in Platelet Concentrates

ABSTRACT Bacterial contamination of platelet concentrates (PCs) still represents an ongoing risk in transfusion-transmitted sepsis. Recently the Pan Genera Detection (PGD) system was developed and FDA licensed for screening of bacterial contamination of PCs directly prior to transfusion. The test principle is based on the immunological detection of lipopolysaccharide (for Gram-negative bacteria) or lipoteichoic acid (for Gram-positive bacteria). In the present study we analyzed the applicability of this method with regard to detection limit, practicability, implementation, and performance. PCs were spiked with Staphylococcus aureus, Bacillus subtilis, and five different Klebsiella pneumoniae strains, as well as eight different Escherichia coli strains. The presence of bacteria was assessed by the PGD immunoassay, and bacteria were enumerated by plating cultures. Application of the PGD immunoassay showed that it is a rapid test with a short hands-on time for sample processing and no demand for special technical equipment and instrument operation. The lower detection limits of the assay for Gram-positive bacteria showed a good agreement with the manufacturer's specifications (8.2 × 103 to 5.5 × 104 CFU/ml). For some strains of K. pneumoniae and E. coli, the PGD test showed analytical sensitivities (>106 CFU/ml) that were divergent from the designated values (K. pneumoniae, 2.0 × 104 CFU/ml; E. coli, 2.8 × 104 CFU/ml). Result interpretation is sometimes difficult due to very faint bands. In conclusion, our study demonstrates that the PGD immunoassay is an easy-to-perform bedside test for the detection of bacterial contamination in PCs. However, to date there are some shortcomings in the interpretation of results and in the detection limits for some strains of Gram-negative bacteria.

[1]  E. Beckers,et al.  Clinical significance of bacteriologic screening in platelet concentrates , 2005, Transfusion.

[2]  E. Beckers,et al.  Effects of skin disinfection method, deviation bag, and bacterial screening on clinical safety of platelet transfusions in the Netherlands , 2006, Transfusion.

[3]  T. Holland-Letz,et al.  Bacterial contamination of platelet concentrates: results of a prospective multicenter study comparing pooled whole blood–derived platelets and apheresis platelets , 2007, Transfusion.

[4]  M. Koppelman,et al.  Development of an internally controlled PCR assay for broad range detection of bacteria in platelet concentrates. , 2008, Journal of microbiological methods.

[5]  M. Arduino,et al.  Growth and endotoxin production of Yersinia enterocolitica and Enterobacter agglomerans in packed erythrocytes , 1989, Journal of clinical microbiology.

[6]  K. Kleesiek,et al.  High-volume extraction of nucleic acids by magnetic bead technology for ultrasensitive detection of bacteria in blood components. , 2007, Clinical chemistry.

[7]  G. Walther-Wenke.  Bacterial contamination of blood components – incidence and significance for homologous and autologous transfusion Bakterielle Kontamination von Blutkomponenten – Inzidenz und Signifikanz für die allogene und autologe Transfusion , 2006 .

[8]  M. Brecher,et al.  Evaluation of BacT/ALERT plastic culture bottles for use in testing pooled whole blood–derived leukoreduced platelet‐rich plasma platelets with a single contaminated unit , 2005, Transfusion.

[9]  Blajchman Ma.  Incidence and significance of the bacterial contamination of blood components. , 2002 .

[10]  R. Benjamin,et al.  Bacterial screening of apheresis platelets and the residual risk of septic transfusion reactions: the American Red Cross experience (2004‐2006) , 2007, Transfusion.

[11]  L. Smith,et al.  Bacterial Contamination of Blood Components , 2003, American Society for Clinical Laboratory Science.

[12]  J. Lupski,et al.  Distribution of repetitive DNA sequences in eubacteria and application to fingerprinting of bacterial genomes. , 1991, Nucleic acids research.

[13]  M. Schmidt,et al.  FACS technology used in a new rapid bacterial detection method , 2006, Transfusion medicine.

[14]  C. Lass‐Flörl,et al.  Prevention of transfusion of platelet components contaminated with low levels of bacteria: a comparison of bacteria culture and pathogen inactivation methods , 2007, Transfusion.

[15]  M. Jacobs,et al.  Detecting bacterial contamination in platelet products. , 2006, Clinical laboratory.

[16]  M. Blajchman,et al.  Improving the bacteriological safety of platelet transfusions. , 2004, Transfusion medicine reviews.

[17]  H. Reesink,et al.  Applications of real-time PCR in the screening of platelet concentrates for bacterial contamination , 2006, Expert review of molecular diagnostics.

[18]  D. Triulzi,et al.  Use of the RQI test for bacterial screening of whole blood platelets. , 2010, American journal of clinical pathology.

[19]  K. Kleesiek,et al.  Novel flow cytometry-based screening for bacterial contamination of donor platelet preparations compared with other rapid screening methods. , 2009, Clinical chemistry.

[20]  K. Triantafilou,et al.  Rough and smooth forms of fluorescein-labelled bacterial endotoxin exhibit CD14/LBP dependent and independent binding that is influencedby endotoxin concentration. , 2000, European journal of biochemistry.

[21]  G. Delage,et al.  Canadian experience with detection of bacterial contamination in apheresis platelets , 2007, Transfusion.

[22]  L. Holness,et al.  Transfusion-transmitted Klebsiella pneumoniae fatalities, 1995 to 2004. , 2006, Transfusion medicine reviews.

[23]  M. Blajchman.  Incidence and significance of the bacterial contamination of blood components. , 2002, Developments in biologicals.

[24]  K. Kleesiek,et al.  Sterility screening of platelet concentrates: questioning the optimal test strategy , 2008, Vox sanguinis.

[25]  S. Lillevang,et al.  Detection of bacterial contamination of platelet components: six years’ experience with the BacT/ALERT system , 2004, Transfusion.

[26]  S. Wagner.  Transfusion‐related bacterial sepsis , 1997, Current opinion in hematology.

[27]  T. Montag,et al.  Basics of flow cytometry–based sterility testing of platelet concentrates , 2006, Transfusion.

[28]  T. Montag,et al.  Methods for the detection of bacterial contamination in blood products , 2008, Clinical chemistry and laboratory medicine.

[29]  M. Brecher,et al.  Transfusion-transmitted bacterial infection. , 1995, Hematology/oncology clinics of North America.

[30]  T. Montag,et al.  Bacteria detection by flow cytometry , 2008, Clinical chemistry and laboratory medicine.

[31]  G. Schreiber,et al.  Transfusion‐transmitted bacterial infectionin the United States, 1998 through 2000 , 2001, Transfusion.

[32]  M. Schmidt,et al.  Comparison of three bacterial detection methods under routine conditions , 2007, Vox sanguinis.

[33]  S. Jonjić,et al.  Protective effect of antilipopolysaccharide monoclonal antibody in experimental Klebsiella infection , 1997, Infection and immunity.

[34]  T. Montag.  Strategies of bacteria screening in cellular blood components , 2008, Clinical chemistry and laboratory medicine.

[35]  B. Tepperman.  Zero tolerance. , 2002, Nursing standard (Royal College of Nursing (Great Britain) : 1987).

[36]  R. Dodd,et al.  Detection of bacterial contamination in apheresis platelet products: American Red Cross experience, 2004 , 2005, Transfusion.

[37]  K. Kleesiek,et al.  Real-time polymerase chain reaction in transfusion medicine: applications for detection of bacterial contamination in blood products. , 2007, Transfusion medicine reviews.

[38]  C. McDonald,et al.  Evaluation of the 3D BacT/ALERT automated culture system for the detection of microbial contamination of platelet concentrates , 2002, Transfusion medicine.

[39]  G. Ortolano,et al.  Detection of bacteria in WBC‐reduced PLT concentrates using percent oxygen as a marker for bacteria growth , 2003, Transfusion.