Evaluation of an automated culture systemfor detecting bacterial contamination of platelets: an analysis with 15 contaminating organisms

Brecher, Mark E.; Means, Norman; Jere, Charles S.; Heath, D. F.; Rothenberg, S.J.; Stutzman, Les C.

doi:10.1046/j.1537-2995.2001.41040477.x

Cited by 133 publications

(140 citation statements)

References 23 publications

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“…Since this and similar organisms account for a large proportion of the bacterial species found to contaminate blood units, such slower growth characteristics need to be considered when optimizing sampling strategies for bacterial detection. 65 Rapid proliferation of bacteria during storage of transfusion products could be decreased if cold temperatures were used. For red cells, this is generally a standard practice and reduces the growth of most bacterial species and the risk of transfusion related sepsis.…”

Section: Prevention Of Bacterial Proliferationrefrigeration Of Platelmentioning

confidence: 99%

“…The manufacturers were encouraged to validate their devices for as many bacterial species as possible and a list of the most common relevant bacteria was agreed on based on literature search, FDA advisory committee input and public workshops input. 10,65 Trials recommended under "actual clinical use" conditions The FDA has proposed that validation of bacterial detection devices for release of platelets for transfusion would require a field trial of the device under actual clinical use conditions. 62 The design of such a trial would be to obtain a sample for bacterial detection by a culture-based device early in the storage period.…”

Section: Current Approval Process For Bacterial Detection Devicesmentioning

confidence: 99%

See 1 more Smart Citation

Bacterial Contamination of Blood Components: Risks, Strategies, and Regulation

Hillyer

Josephson²,

Blajchman³

et al. 2003

Hematology

221

210

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Bacterial contamination of transfusion products, especially platelets, is a longstanding problem that has been partially controlled through modern phlebotomy practices, refrigeration of red cells, freezing of plasma and improved materials for transfusion product collection and storage. Bacterial contamination of platelet products has been acknowledged as the most frequent infectious risk from transfusion occurring in approximately 1 of 2,000-3,000 whole-blood derived, random donor platelets, and apheresis-derived, single donor platelets. In the US, bacterial contamination is considered the second most common cause of death overall from transfusion (after clerical errors) with mortality rates ranging from 1:20,000 to 1:85,000 donor exposures. Estimates of severe morbidity and mortality range from 100 to 150 transfused individuals each year.Concern This Joint ASH and AABB Educational Session reviews the risks, testing strategies, and regulatory approaches regarding bacterial contamination of blood components to aid in preparing practitioners of hematology and transfusion medicine in understanding the background and clinical relevance of this clinically important issue and in considering the approaches currently available for its mitigation, as well as their implementation.In this chapter, Drs. Hillyer and Josephson review the background and significance of bacterial contamination, as well as address the definitions, conceptions and limitations of the terms risk, safe and safety. They then describe current transfusion risks including non-infectious serious hazards of transfusion, and current and emerging viral risks. In the body of the text, Dr. Blajchman reviews the prevalence of bacterial contamination in cellular blood components in detail with current references to a variety of important studies. He then describes the signs and symptoms of transfusion-associated sepsis and the sources of the bacterial contamination for cellular blood products including donor bacteremia, and contamination during whole blood collection and of the collection pack. This is followed by strategies to decrease the transfusion-associated morbidity/mortality risk of contaminated cellular blood products 576American Society of Hematology including improving donor skin disinfection, removal of first aliquot of donor blood, pretransfusion detection of bacteria, reducing recipient exposure, and pathogen reduction/inactivation. In the final sections, Drs. Vostal, Epstein and Goodman describe the regulations and regulatory approaches critical to the appropriate implementation of a bacterial contamination screening and limitation program including their and/or the FDA's input on prevention of bacterial contamination, bacterial proliferation, and detection of bacteria in transfusion products. This is followed by a discussion of sampling strategy for detection of bacteria in a transfusion product, as well as the current approval process for bacterial detection devices, trials recommended under "actual clinical use" conditions, pathogen reduction ...

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Section: Prevention Of Bacterial Proliferationrefrigeration Of Platelmentioning

confidence: 99%

Section: Current Approval Process For Bacterial Detection Devicesmentioning

confidence: 99%

Bacterial Contamination of Blood Components: Risks, Strategies, and Regulation

Hillyer

Josephson²,

Blajchman³

et al. 2003

Hematology

221

210

View full text Add to dashboard Cite

show abstract

“…For platelet transfusions, Ͼ30% of clinical complications are due to Staphylococcus species, particularly Staphylococcus epidermidis and S. aureus, but numerous other bacterial species may also be responsible (5,31,38). Conventional methods for the detection of these bacteria in blood components involve culturing and identification by morphological, biochemical, and immunological characteristics.…”

mentioning

confidence: 99%

Two Novel Real-Time Reverse Transcriptase PCR Assays for Rapid Detection of Bacterial Contamination in Platelet Concentrates

2004

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The incidence of platelet bacterial contamination is approximately 1 per 2,000 units and has been acknowledged as the most frequent infectious risk from transfusion. In preliminary studies, the sterility of platelet concentrates (PCs) was tested with an automated bacterial blood culturing system and molecular genetic assays. Two real-time reverse transcriptase PCR (RT-PCR) assays performed in a LightCycler instrument were developed and compared regarding specificity and sensitivity by the use of different templates to detect the majority of the clinically important bacterial species in platelets. Primers and probes specific for the conserved regions of the eubacterial 23S rRNA gene or the groEL gene (encoding the 60-kDa heat shock protein Hsp60) were designed. During the development of the 23S rRNA RT-PCR, problems caused by the contamination of reagents with bacterial DNA were noted. Treatment with 8-methoxypsoralen and UV irradiation reduced the level of contaminating DNA. The sensitivity of the assays was greatly influenced by the enzyme system which was used. With rTth DNA polymerase in a one-enzyme system, we detected 500 CFU of Escherichia coli or Staphylococcus epidermidis/ml. With a two-enzyme system consisting of Moloney murine leukemia virus RT and Taq DNA polymerase, we detected 16 CFU/ml. With groEL mRNA as the target of RT-PCR under optimized conditions, we detected 125 CFU of E. coli/ml, and no problems with false-positive results caused by reagent contamination or a cross-reaction with human nucleic acids were found. Furthermore, the use of mRNA as an indicator of viability was demonstrated. Here we report the application of novel real-time RT-PCR assays for the detection of bacterial contamination of PCs that are appropriate for transfusion services.

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“…Long incubation periods, however, are necessary, in particular for PCs with low bacterial concentrations and slowly growing strains. The detection of P. acnes by the automated blood culture method requires more than 4 days (7,9), and the detection of Mycobacterium is reported to require 20 days (25).…”

Section: Discussionmentioning

confidence: 99%

Rapid Screening Method for Detection of Bacteria in Platelet Concentrates

Ribault¹,

Harper²,

Grave³

et al. 2004

J Clin Microbiol

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Public awareness has long focused on the risks of the transmission of viral agents through blood product transfusion. This risk, however, pales in comparison to the less publicized danger associated with the transfusion of blood products contaminated with bacteria, in particular, platelet concentrates. Up to 1,000 cases of clinical sepsis after the transfusion of platelet concentrates are reported annually in the United States. The condition is characterized by acute reaction symptoms and the rapid onset of septicemia and carries a 20 to 40% mortality rate. The urgent need for a method for the routine screening of platelet concentrates to improve patient safety has long been recognized. We describe the development of a rapid and highly sensitive method for screening for bacteria in platelet concentrates for transfusion. No culture period is required; and the entire procedure, from the time of sampling to the time that the final result is obtained, takes less than 90 min. The method involves three basic stages: the selective removal of platelets by filtration following activation with a monoclonal antibody, DNA-specific fluorescent labeling of bacteria, and concentration of the bacteria on a membrane surface for enumeration by solid-phase cytometry. The method offers a universal means of detection of live, nondividing, or dead gram-negative and gram-positive bacteria in complex cellular blood products. The sensitivity is higher than those of the culture-based methods available at present, with a detection limit of 10 to 10 2 CFU/ml, depending upon the bacterial strain.

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Evaluation of an automated culture systemfor detecting bacterial contamination of platelets: an analysis with 15 contaminating organisms

Cited by 133 publications

References 23 publications

Bacterial Contamination of Blood Components: Risks, Strategies, and Regulation

Bacterial Contamination of Blood Components: Risks, Strategies, and Regulation

Two Novel Real-Time Reverse Transcriptase PCR Assays for Rapid Detection of Bacterial Contamination in Platelet Concentrates

Rapid Screening Method for Detection of Bacteria in Platelet Concentrates

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