Rapid Screening Method for Detection of Bacteria in Platelet Concentrates

Ribault, Sébastien; Harper, Kirsty; Grave, Linda; Lafontaine, C.; Nannini, P.; Raimondo, A.; Faure, I. Besson

doi:10.1128/jcm.42.5.1903-1908.2004

Cited by 40 publications

(35 citation statements)

References 25 publications

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“…Since platelets should be stored at room temperature that makes them an excellent growth medium for bacteria, it is declared as a major difficulty in transfusion medicine [2]. The transfusion risk of a bacterial contaminated platelet concentrates (PC) is higher than viral pathogen such as HIV, HBV, HCV and HTLV [3]. Bacterial contamination is considered the third most common cause of death overall from transfusion reported to the US Food and Drug Association (FDA), following transfusion-related acute lung injury and hemolytic reactions.…”

Section: Background and Objectivesmentioning

confidence: 99%

Evaluation of the Sensitivity and Specificity of Use of Glucose and pH for Bacterial Screening of Platelet Concentrates Compared to the Bact/Alert

Razjou

Naghadeh

Ferdowsi

et al. 2016

Indian J Hematol Blood Transfus

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Bacterial contamination of blood components is the major infectious risk in transfusion medicine. Since platelets should be stored at room temperature that makes them an excellent growth medium for bacteria; it is mentioned as a major problem in transfusion medicine. Transfusion risk of a bacterial contaminated platelet concentrate is higher than viral pathogen such as HIV, HBV, HCV and HTLV. The objective of this study was to evaluation of the sensitivity and specificity of use of glucose and pH for bacterial screening of platelet concentrates compared to the Bact/Alert. 1332 platelet concentrates were screened by the Bact/Alert system for aerobic and anaerobic bacterial contamination. Bacterial contamination was also evaluated by using urine reagent strips (Multistix10 SG Bayer) and culture methods. Moreover PH screening with a pH meter (Metrohm 744 Swiss) and glucose was also used for detection of bacterial contamination. The rate of bacterial contamination detected by the Bact/Alert system in platelet concentrates was 25 in 1332 (1.9 %). It contained 15 (1.1 %) for aerobic bacteria and 10 (.8 %) for anaerobic bacteria. 226 of 1332 were considered as containing bacteria by using urine reagent strips. Six of the 226 units were also positive by the Bact/Alert system. Three of those units were culture positive for aerobic bacteria and three for anaerobic. The result of platelet concentrates that underwent pH screening by use of pH meter and a pH portion of urine reagent strips was the same. The sensitivity and specificity of considering glucose alone for detection of bacterial contamination were 12 and 98 % respectively. For pH alone, these were 24 and 83 %. For glucose and/or pH, these were 24 and 83 %; and for combination of glucose and pH, these were 12 and 98 %. Our results showed use of glucose/pH strips would improve the safety of blood products and should be encouraged.

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Section: Background and Objectivesmentioning

confidence: 99%

Evaluation of the Sensitivity and Specificity of Use of Glucose and pH for Bacterial Screening of Platelet Concentrates Compared to the Bact/Alert

Razjou

Naghadeh

Ferdowsi

et al. 2016

Indian J Hematol Blood Transfus

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“…In a 20-min incubation period in the agglutination solution, more than 99% of the RBCs compacted into a pellet. Residual RBCs could then be removed by filtration and the cell free supernatant analyzed by the solid-phase scanning cytometry system (25). Very low levels of bacteria can be concentrated from a large test sample volume by filtration, giving a significant advantage over plate culture methods.…”

Section: Discussionmentioning

confidence: 99%

“…The use of a solid-phase cytometer has already been described as a highly sensitive bacterial screening system for various biologic products, including platelet concentrates (2,19,25; P. Morel, M. Dechaseaux, X. Bertrand, C. Naegelen, and D. Talon, Transfusion 43:SP13, 2003). When adapting the method for use with RBCC, a lysis step to remove red blood cells was tested; however, the free hemoglobin generated too high a background of fluorescence.…”

Section: Discussionmentioning

confidence: 99%

“…The sample was then filtered through a 0.4-m black membrane (Whatman), which retains the bacteria on its surface. The black membrane was transferred to the Scansystem solid-phase cytometer, and the number of labeled bacteria was determined (25).…”

Section: Propagationmentioning

confidence: 99%

“…The system, which was described previously for detection of bacteria in platelet concentrates (25), includes an erythrocyte removal step based on the induction of red cell agglutination. In the subsequent bacterial labeling of the erythrocyte-depleted sample, the membranes of gram-positive and gram-negative bacteria are permeabilized, allowing the entry and DNA-specific binding of a fluorophore marker.…”

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confidence: 99%

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Detection of Bacteria in Red Blood Cell Concentrates by the Scansystem Method

Ribault¹,

Faucon²,

Grave³

et al. 2005

J Clin Microbiol

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Bacterial contamination remains one of the major risks associated with blood product transfusion. The kinetics of bacterial growth in red blood cell concentrates (RBCC) is different than otherwise due to storage at 4°C, conditions in which most bacteria do not survive. Psychrophilic bacteria such as Yersinia enterocolitica, however, can proliferate from a very low level of contamination to clinically significant levels at 4°C and are known to cause severe transfusion-related infections. A screening method allowing the early detection of very low levels of bacteria in RBCC would improve transfusion safety. The Scansystem method has been previously described for detection of bacteria in platelet concentrates. We present here a modification of the system for detection of low levels of bacteria in RBCC. The Scansystem RBC kit protocol requires three steps, i.e., the agglutination and selective removal of RBCs, a labeling stage during which bacteria are labeled with a DNA-specific fluorophore, and finally recovery of bacteria on the surface of a black membrane for analysis using the Scansystem. The entire procedure from sampling to result can be completed in 90 min. Both gram-negative and gram-positive bacteria in RBCC are detected with a higher sensitivity than with currently available culture-based methods. The Scansystem RBC kit is shown to be sensitive enough to identify low-level bacterial contamination in a single unit tested in a pool of up to 20 RBCC samples (detection limit of between 1 and 10 CFU/ml depending on the bacterial strain). The method therefore lends itself to incorporation into high-sample-throughput screening programs.Due to the availability of highly sensitive virus screening kits, in donated blood products bacterial contamination, now the greatest risk for patients receiving a transfusion, is found 50 to 250 times more frequently than viral infection such as human immunodeficiency virus, hepatitis virus, or human T-cell lymphotropic virus infection. Transfusion-related bacterial infections are usually associated with a rapid onset of sepsis with high mortality rates (4, 16). Platelet concentrate preparations are known to carry a greater risk of bacterial infections, as they are required to be stored at 20 to 24°C (3, 6, 7, 10). With the significantly higher number of red blood cell concentrates (RBCC) transfused, however, the incidence of transfusionassociated bacterial contamination is similar (5,11,18,28). In the French Bacthem case-control study, 41 bacterium-related transfusion reactions were reported during a 2-year period, including 25 following transfusion with RBCC, with 4 fatalities, and 16 following platelet transfusion, with 2 fatalities (23).Currently, the examination of the RBCC bag for evidence of hemolysis just prior to transfusion is the only screening method carried out to check for bacterial contamination (12). However, visual inspection is subjective, and hemolysis also occurs in noncontaminated units nearing the end of their shelf life. Identification of low levels of bact...

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