Study on platelet components production in 19 provincial blood centers in China before and during the COVID-19 epidemic / 中国输血杂志

【Objective】 To study the changes of platelet components(PC), apheresis platelets (AP) and pooled platelet concentrates (PPC) production of 19 provincial blood centers before and during the COVID-19 epidemic. 【Methods】 The data related to the collection of AP and the preparation of PPC from 2016 to 2021 of 19 provincial blood centers was collected. The production of PC, AP and PPC during the four years before the epidemic (i.e. 2016-2019) and during the COVID-19 epidemic (i.e. 2020 and 2021) were calculated respectively, and the change of production was analyzed. 【Results】 The total production of PC in 19 blood centers steadily increased from 2016 to 2019, with a decrease of 4.16% in 2020 and an increase of 15.60% in 2021, exceeding the output before the COVID-19 epidemic. In 2020, the production of PC of 42.11% (8/19) blood centers decreased compared with 2019, while 94.74% (18/19) in 2021 increased compared with 2020. The changes of AP output was basically consistent with the trend of PC. The total production of PPC in 2017 and 2018 both doubled compared to the previous year, while decreased by 67.98% in 2019, increased by 30.38% in 2020 and decreased by 27.08% in 2021. 【Conclusion】 The total production of PC kept increasing steadily between 2016 and 2019, but decreased in 2020 under the COVID-19 epidemic, with some blood centers being significantly affected. In 2021, with the strong support from government and various measures by blood centers, the total production of PC increased.

Assessment of Qualitative and Quantitative Changes in 5 days stored Platelet Concentrates in Bangabandhu Sheikh Mujib Medical University

Background: The fast-growing demand for platelet concentrates (PC) necessitates the storage of these blood products before transfusion. Platelets are prepared as concentrates from the whole blood or by plateletpheresis. Qualitative and quantitative assessment of these PCs is an important issue in transfusion medicine. To assess the qualitative, quantitative changes and bacteriological safety of 5 days of stored platelet concentrates (PC).Material & Methods:This prospective study was conducted at the department of Clinical Pathology in collaboration with the Department of Transfusion Medicine, Bangabandhu Sheikh Mujib Medical University (BSMMU), Dhaka from April 2008 to April 2009. A total of 65 healthy donors were included in the study as per the inclusion and exclusion criteria. Therefore, 65 platelet concentrates (bags/units) were prepared from the donors. Purposive sampling of the units was done. pH and platelet indices (PLT, MPV, PDW and P-LCR) were measured and Gram staining of PCs was performed on days 0 and 5. Statistical significant tests were done at a 95% confidence interval using the statistical package for social science (SPSS).Results:The mean (±SD) pH was 7.18±0.07 ranging from 7.0 to 7.3 during day 0. On day 5 the mean (±SD) pH was 6.77±0.11 and their range was from 6.5 to 7. The mean pH difference was statistically significant (p<0.05) between day 0 and day 5. The mean (±SD) PLT/unit was 70.56±15.56 x109/unit and it ranged from 38.01 to 110.6 x109/unit during day 0. On day 5 the mean (±SD) PLT/unit level was 68.46±15.52 x109/unit and it ranged from 36.82 to 107.2 x109/unit. The mean PLT/unit difference was statistically significant (p<0.05) between day 0 and day 5. The mean (±SD) MPV was 9.34±0.92 fl and it ranged from 7.5 to 11.5 fl during day 0. During day 5 the mean (±SD) MPV was 9.27±0.99 fl ranging from 7.0 to 11.2 fl. The mean (±SD) PDW was 10.07±1.61 fl and which ranged from 7.4 to 14.4 fl during day 0. During day 5 the mean (±SD) PDW was 10.72±1.71 fl ranging from 7.0 to 15.4 fl. The mean (±SD) PLCR was 18.28±5.67 % and it ranged from 8.0 to 32.5 % during day 0. During day 5 the mean (±SD) PLCR was 21.18±5.91 % and it ranged from 10.0 to 36.3 %. The mean PLT, PDW and PLCR differences were statistically significant (p<0.05) between day 0 and day 5 in the unpaired t-test, however, the mean MPV difference was not statistically significant (p<0.05) between day 0 and day 5. Gram staining of platelet concentrates on day 0 and day 5 found no bacteria.Conclusions:Storage-induced lesions take place in PCs when stored for 5 days in second-generation storage containers under the currently recommended conditions, but how far these changes are clinically relevant needs to be investigated.

Assessment of Qualitative and Quantitative Changes in 5 days stored Platelet Concentrates in a tertiary care Hospital of Bangladesh

Background: The fast growing demand for platelet concentrates (PC) necessitates the storage of these blood products prior to transfusion. Platelets are prepared as concentrates from the whole blood or by plateletpheresis. Qualitative and quantitative assessment of these PCs are an important issue in transfusion medicine. Aim of the study: To assess the qualitative, quantitative changes and bacteriological safety of 5 days stored platelet concentrates (PC).Material & Methods:This prospective study was conducted at the department of Clinical Pathology in collaboration with the department of Transfusion medicine, Bangabandhu Sheikh Mujib Medical University (BSMMU), Dhaka during April 2008 to April 2009. A total of 65 healthy donors were included for the study as per the inclusion and exclusion criteria. Therefore, 65 platelet concentrates (bags/units) were prepared from the donors. Purposive sampling of the units was done. pH and platelet indices (PLT, MPV, PDW and P-LCR) were measured and Gram staining of PCs were performed on day 0 and 5. Statistical significant tests were done at 95% confidence interval using statistical package for social science (SPSS).Results:The mean (±SD) pH was 7.18±0.07 ranging from 7.0 to 7.3 during day 0. During day 5 the mean (±SD) pH was 6.77±0.11 and their range was from 6.5 to 7. The mean pH difference was statistically significant (p<0.05) between day 0 and day 5. The mean (±SD) PLT/unit was 70.56±15.56 x109/unit and it ranged from 38.01 to 110.6 x109/unit during day 0. During day 5 the mean (±SD) PLT/unit level was 68.46±15.52 x109/unit and it ranged from 36.82 to 107.2 x109/unit. The mean PLT/unit difference was statistically significant (p<0.05) between day 0 and day 5. The mean (±SD) MPV was 9.34±0.92 fl and it ranged from 7.5 to 11.5 fl during day 0. During day 5 the mean (±SD) MPV was 9.27±0.99 fl ranging from 7.0 to 11.2 fl. The mean (±SD) PDW was 10.07±1.61 fl and which ranged from 7.4 to 14.4 fl during day 0. During day 5 the mean (±SD) PDW was 10.72±1.71 fl ranging from 7.0 to 15.4 fl. The mean (±SD) PLCR was 18.28±5.67 % and it ranged from 8.0 to 32.5 % during day 0. During day 5 the mean (±SD) PLCR was 21.18±5.91 % and it ranged from 10.0 to 36.3 %. The mean PLT, PDW and PLCR difference were statistically significant (p<0.05) between day 0 and day 5 in unpaired t-test, however the mean MPV difference was not statistically significant (p<0.05) between day 0 and day 5. Gram staining of platelet concentrates on day 0 and day 5 found no bacteria.Conclusions:Storage-induced lesions take place in PCs, when stored for 5 days in second generation storage containers under the currently recommended conditions, but how far these change are clinically relevant need to be investigated

Autologous platelet concentrates for facial rejuvenation

Abstract Autologous platelet concentrates (APCs) are promising therapeutic agents in facial rejuvenation since they are a great source of cytokines, growth factors and other biologically active substances. Obtained from the patient's blood, they have the advantages of reducing immunological reactions, making the procedure safer, well tolerated, with minimal adverse effects and lower cost. Currently, they are used for facial rejuvenation both in combination with microneedling and in mesotherapy techniques, as well as to treat facial acne scars, melasma and wounds after laser ablative treatments. This review summarizes current knowledge on the use of APCs, ranging from basic concepts related to their composition and mechanisms of action to up-to-date information on their clinical efficacy. Methodology MEDLINE (PubMed) was searched from inception through 2021 for English language publications on APCs for facial rejuvenation. Results A total of 100 files were found. Based on the available literature, APCs for skin rejuvenation are safe and well tolerated. The most studied product is the first-generation material, platelet-rich plasma (PRP). Conclusions The results are in general favorable, but the quality of the studies is low. The second and third generation products, platelet-rich fibrin (PRF) and injectable platelet-rich fibrin (i-PRF), respectively, are easier to be obtained and, at least in vitro , seem to induce greater collagen production than PRP, especially under lower relative centrifugation forces, but to date only a few clinical trials evaluating these products exist. More high-quality trials with appropriate follow-up are necessary to provide adequate evidence that may help to improve the treatment regimens with APCs. Many aspects should be considered when designing clinical trials to evaluate APCs, such as the patients' characteristics that best predict a favorable response, the optimal number of sessions and the interval between them, the characteristics of the studies and the development of better instruments to evaluate skin aging.

Optimized broad-range real-time PCR-based method for bacterial screening of platelet concentrates / Otimização de um método baseado em PCR em tempo real para rastreamento de bactérias em concentrados de plaquetas

Abstract Bacterial contamination of blood components remains a major challenge in transfusion medicine, particularly, platelet concentrates (PCs) due to the storage conditions that support bacterial proliferation. In this study, we develop a rapid, sensitive and specific real-time PCR protocol for bacterial screening of PCs. An internally controlled real-time PCR-based method was optimized and validated with our proprietary 16S Universal PCR Master Mix (IBMP/Fiocruz), which targets a conserved region of the bacterial 16S rRNA gene. Nonspecific background DNA was completely eliminated by treating the PCR Master Mix with ethidium monoazide (EMA). A lower limit of detection was observed for 10 genome equivalents with an observed Ct value of 34±1.07 in calibration curve generated with 10-fold serial dilutions of E. coli DNA. The turnaround time for processing, including microbial DNA purification, was approximately 4 hours. The developed method showed a high sensitivity with no non-specific amplification and a lower time-to-detection than traditional microbiological methods, demonstrating it to be an efficient means of screening pre-transfusion PCs.

Resumo A contaminação bacteriana dos componentes sanguíneos é um grande desafio na medicina transfusional, principalmente nos concentrados de plaquetas (PCs) devido às condições de armazenamento que favorecem a proliferação bacteriana. Neste estudo, desenvolvemos um protocolo de PCR em tempo real rápido, sensível e específico para a triagem bacteriana de PCs. Um método baseado em PCR em tempo real, controlado internamente, foi otimizado e validado com um Master Mix Universal PCR 16S (IBMP / Fiocruz), que detecta uma região conservada do gene 16S rRNA bacteriano. O background de DNA não específico foi completamente eliminado tratando a PCR Master Mix com monoazida de etídio (EMA). O limite de detecção inferior observado foi de 10 cópias equivalentes do genoma com um valor de Ct 34 ± 1,07, a curva de calibração foi gerada com diluições seriada de 10 vezes do DNA de E. coli. O tempo de processamento, incluindo a purificação microbiana do DNA, foi de aproximadamente 4 horas. O método desenvolvido mostrou alta sensibilidade sem amplificação inespecífica e menor tempo de detecção do que os métodos microbiológicos tradicionais, demonstrando ser um meio eficiente de triagem de PCs pré-transfusionais.

Efectividad de los concentrados plaquetarios en el tratamiento de los desórdenes temporomandibulares / Effectiveness of platelet concentrates in the treatment of temporomandibular disorders

Introducción: Los desórdenes temporomandibulares son un grupo de trastornos que afectan la articulación temporomandibular y/o los tejidos musculoesqueléticos asociados. Objetivo: Determinar la efectividad de los concentrados de plaquetas en el tratamiento de los desórdenes temporomandibulares. Métodos: La búsqueda de la literatura fue realizada desde enero del 2014 hasta abril del 2019, en las bases de datos biomédicas: PubMed, Embase, SciELO, Scopus, Science Direct, Sistema de información sobre literatura gris en Europa, Literatura Latinoamericana y del Caribe en Ciencias de la Salud, Google Académico y el Registro Central de Ensayos Clínicos Cochrane. Se definieron como criterios de selección de los estudios que fueran ensayos clínicos aleatorizados, con una antigüedad máxima de cinco años, que reportarann la efectividad (reducción del dolor y aumento de apertura máxima) de los concentrados plaquetarios en el tratamiento de los desórdenes temporomandibulares. El riesgo de sesgo de los estudios fue analizado por medio del Manual Cochrane de revisiones sistemáticas de intervenciones. Resultados: La estrategia de búsqueda resultó en nueve artículos, de los cuales el 100 por ciento reportó que no había diferencia en la reducción del dolor y el aumento de apertura máxima de los concentrados plaquetarios en el tratamiento de los desórdenes temporomandibulares. Conclusiones: La literatura revisada sugiere que existe una ligera evidencia de los beneficios potenciales de las inyecciones intraarticulares de los concentrados plaquetarios en pacientes con desórdenes temporomandibulares. Sin embargo, es necesario establecer un protocolo estandarizado para la preparación y aplicación de estos concentrados(AU)

Introduction: Temporomandibular disorders are a group of dysfunctions which affect the temporomandibular joint and/or associated musculoskeletal tissues. Objective: Determine the effectiveness of platelet concentrates in the treatment of temporomandibular disorders. Methods: A bibliographic search was conducted from January 2014 to April 2019 in the biomedical databases PubMed, Embase, SciELO, Scopus, Science Direct, System for Information on Gray Literature in Europe, Latin American and Caribbean Health Sciences Literature, Google Scholar, and Cochrane Central Register of Clinical Trials. The following selection criteria were defined for the studies: randomized clinical trials published in the last five years and reporting on the effectiveness (pain reduction and maximum opening increase) of platelet concentrates in the treatment of temporomandibular disorders. Bias risk analysis was based on the Cochrane manual of systematic reviews of interventions. Results: Nine papers were retrieved, of which 100 percent reported no differences in pain reduction or maximum opening increase resulting from the use of platelet concentrates in the treatment of temporomandibular disorders. Conclusions: The literature review conducted suggests that there is slight evidence of the potential benefits of intra-articular injections of platelet concentrates in patients with temporomandibular disorders. However, a standardized protocol should be established for the preparation and application of these concentrates(AU)

PLATELET CONCENTRATES FOR PERIODONTAL REGENERATION

@#Platelet concentrates are derivatives of blood that aid in haemostasis and wound healing after periodontal regenerative procedures. Its ability to act as a natural scaffold of growth factors has gained significance in many surgical procedures. This narrative review discusses the different platelet concentrates, their centrifugation protocols, advantages and disadvantages and their application in periodontal regenerative procedures. An electronic search of PubMed or MEDLINE was conducted for relevant material from the published literature up to 2020. The key words looked for were “Platelet concentrates, Platelet rich plasma, Platelet rich fibrin and periodontal regeneration.” We have used the filters comparative human studies, animal studies, randomized controlled trials, case reports and systematic reviews. The searches were limited to articles in English language and articles describing platelet concentrates and its relation to periodontal regeneration were collected and used to prepare a concise review.

Quality Parameters of Platelet Concentrates Collected from Manual and Automated Blood Collection Mixer

Abstract@#Introduction: Inadequate mixing during the blood collection process might affect the quality of platelet concentrates (PCs). Currently, two different mixing methods are used during whole blood collection: manual mixing and mixing using an automated blood collection mixer. However, the cost between manual and automated blood collection mixer differed largely and pose a dilemma for a blood transfusion service. The objective of this study was to evaluate PCs quality using either manual mixing or automated procedure. Methods: This was a cross-sectional study conducted at the Advanced Medical and Dental Institute, Universiti Sains Malaysia. Thirty eligible participants aged 20 to 45 were included in this study, and a unit of 450 mL whole blood was collected from each participant. Fifteen units of whole blood were mixed by an automated blood collection mixer and the other 15 units were mixed using the manual mixing. All PCs were produced from platelet-rich plasma and stored at 20–24°C for 5 days. Quality parameters such as platelet count, leucocyte count, and pH were measured for each PCs on day 1 and day 5. Results: Platelet count on day 1 was significantly higher than on day 5 (p = 0.01) for both mixing methods. There was no statistically significant difference in any of the PCs quality parameters between the two types of mixing methods at either day 1 or day 5 of storage (p > 0.05). Conclusion: Comparable PCs quality is achieved from both manual mixing and automated procedures.

Current application of concentrated growth factors in periodontal tissue regeneration / 口腔疾病防治

@#As a new generation of autologous platelet concentrates, concentrated growth factors (CGF) have demonstrated potential for promoting soft and hard tissue regeneration in vitro and in vivo, and this potential has also been demonstrated in clinical trials. CGF may be used alone or with bone graft materials in guided tissue regeneration (GTR), bone regeneration, internal sinus elevation, and reconstruction of bone defects after the removal of jaw cysts. As a rich source of growth factors, CGF have performed well in periodontal regeneration.

Análisis estructural de la fibrina rica en plaquetas y sus aplicaciones en odontología regenerativa / Structural Analysis of Platelet-Rich Fibrin and its Applications in Regenerative Dentistry

Antecedentes: La fibrina rica en plaquetas (PRF) es un concentrado plaquetario que se está usando con mayor frecuencia en medicina y odontología. Los resultados clínicos son variables posiblemente porque hay diferentes protocolos de obtención, equipos de centrifugado y técnicas de colocación. El desconocimiento de los aspectos estructurales puede afectar el resultado clínico. Objetivo: Describir las características estructurales de la PRF en las diferentes zonas de la membrana. Métodos: Se realizó un estudio experimental in vitro con 15 muestras de sangre periférica tomada de cinco voluntarios adultos, sanos, asistentes a la clínica odontológica de la Universidad Antonio, Popayán. Se hizo hemograma inicial, se recolectó sangre y se centrifugó (10 min x 3000 rpm). Las muestras se analizaron histológicamente y con microscopía electrónica de barrido (SEM). Se describió la estructura de la fibrina, las plaquetas y los leucocitos. Resultados: El promedio de recuento de plaquetas en sangre total fue de 251±31,74 x103 x mm3 y en PRF fue de 832±123,43 x103 x mm3. Macroscópicamente, se identificaron tres zonas del PRF: una superior con pocas plaquetas, una zona leucocitaria (BC) y una zona corpuscular roja. En el análisis de microscopía óptica muestra que en la zona BC hay mayor concentración plaquetaria. El análisis por SEM comprueba que la estructura de la red de fibrina y el contenido celular son diferenciales en cada zona. Conclusión: A partir del conocimiento estructural del PRF se pueden proponer aplicaciones que mejoren el rendimiento del material y por tanto los resultados clínicos.

Background: Platelet rich fibrin (PRF) is a platelet concentrate that is used most frequently in medicine and dentistry. Clinical results are variables because there are different protocols, centrifugation equipment and placement techniques. Ignorance of structural elements can affect the clinical outcome. Purpose: To describe the structural characteristics of Platelet Rich Fibrin in the different zones of the membrane. Methods: Experimental in vitro study with 15 blood samples taken from five healthy adult volunteers attending the dental clinic of the University Antonio Nariño, Popayán. The basal blood count was made, then blood was collected and centrifuged (10 minutes x 3000 rpm). The samples were analyzed histologically and with scanning electron microscopy (SEM). The structure of fibrin, platelets and leukocytes was described. Results: The average platelet count in whole blood was 251 ± 31.74 x103 x mm3 and in PRF it was 832±123.43 x103 x mm3. Macroscopically, three areas of the PRF were identified: an upper one with few platelets (FPP), a buffy coat (BC) and the corpuscular network zone (RBC). In the analysis of optical microscopy shows that in the BC area there is a higher platelet concentration. The SEM analysis proves that the structure of the fibrin network and cellular content is differential in each zone. Conclusion: From the structural knowledge of the PRF, applications that may be improve the performance of the biomaterial and therefore the clinical results can be proposed.

Optimized broad-range real-time PCR-based method for bacterial screening of platelet concentrates

Abstract Bacterial contamination of blood components remains a major challenge in transfusion medicine, particularly, platelet concentrates (PCs) due to the storage conditions that support bacterial proliferation. In this study, we develop a rapid, sensitive and specific real-time PCR protocol for bacterial screening of PCs. An internally controlled real-time PCR-based method was optimized and validated with our proprietary 16S Universal PCR Master Mix (IBMP/Fiocruz), which targets a conserved region of the bacterial 16S rRNA gene. Nonspecific background DNA was completely eliminated by treating the PCR Master Mix with ethidium monoazide (EMA). A lower limit of detection was observed for 10 genome equivalents with an observed Ct value of 34±1.07 in calibration curve generated with 10-fold serial dilutions of E. coli DNA. The turnaround time for processing, including microbial DNA purification, was approximately 4 hours. The developed method showed a high sensitivity with no non-specific amplification and a lower time-to-detection than traditional microbiological methods, demonstrating it to be an efficient means of screening pre-transfusion PCs.

Resumo A contaminação bacteriana dos componentes sanguíneos é um grande desafio na medicina transfusional, principalmente nos concentrados de plaquetas (PCs) devido às condições de armazenamento que favorecem a proliferação bacteriana. Neste estudo, desenvolvemos um protocolo de PCR em tempo real rápido, sensível e específico para a triagem bacteriana de PCs. Um método baseado em PCR em tempo real, controlado internamente, foi otimizado e validado com um Master Mix Universal PCR 16S (IBMP / Fiocruz), que detecta uma região conservada do gene 16S rRNA bacteriano. O background de DNA não específico foi completamente eliminado tratando a PCR Master Mix com monoazida de etídio (EMA). O limite de detecção inferior observado foi de 10 cópias equivalentes do genoma com um valor de Ct 34 ± 1,07, a curva de calibração foi gerada com diluições seriada de 10 vezes do DNA de E. coli. O tempo de processamento, incluindo a purificação microbiana do DNA, foi de aproximadamente 4 horas. O método desenvolvido mostrou alta sensibilidade sem amplificação inespecífica e menor tempo de detecção do que os métodos microbiológicos tradicionais, demonstrando ser um meio eficiente de triagem de PCs pré-transfusionais.

Antimicrobial susceptibility profile and research of mec A and erm genes in coagulase-negative staphylococci isolated from platelet concentrates bags

Abstract In recent years, several studies have described the clinical impact of bacterial infection associated with transfusion of platelet concentrates (PCs). Among the blood components, PCs are responsible for the highest rates of bacterial contamination as well as septic transfusion reactions. We assessed antimicrobial susceptibility profile, resistance to methicillin (MRCoNS), and resistance to macrolides, lincosamides and streptogramins of group B (MLSB) of 16 coagulase-negative staphylococci (CoNS) isolates from an investigation in 691 PCs bags. We then compared conventional and automated phenotypic methods, disc diffusion test (DD) and VITEK(r) 2, respectively as well as phenotypic and genotypic methods (Polymerase Chain Reaction - PCR). All CoNS were susceptible to vancomycin. The disc diffusion test characterized 18.75% as MRCoNS and 37.5% with inducible resistance to MLSB (iMLSB), and with VITEK(r) 2, 6.3% and 31.25%, respectively. The mecA gene was detected in 18.75% and the erm gene in 31.25% of the isolates. In this study, we found equal percentage values between presence of the mecA gene by PCR and resistance to methicillin using cefoxitin by DD test, evidence of the erm gene by PCR, and iMLSB resistance by automation (VITEK(r) 2). Moreover, we identified three strains with beta-lactamase overproduction, and the occurrence of a bigger mistake was verified when automation was compared with DD test. And we observed that D-test was the most reliable for the detection of iMLSB resistance in Staphylococcus sp.

Fibrina rica en plaquetas (FRP): Una alternativa terapéutica en odontología

El objetivo de esta revisión, fue describir de una manera sencilla las principales características de la Fibrina Rica en Plaquetas (FRP), su composición, propiedades y aplicación clínica. La FRP, biomaterial autógeno y concentrado plaquetario de segunda generación, es una matriz de fibrina que contiene leucocitos, plaquetas y factores de crecimiento, que son necesarios para los procesos de cicatrización, lo que brinda a este biomaterial, una gran utilidad en diversas áreas de la salud, incluyendo la odontología. Con esta revisión concluimos que la FRP es una alternativa real para mejorar la cicatrización de procedimientos quirúrgicos y potenciar otros biomateriales regenerativos en diversas áreas de la odontología, además de su accesibilidad y bajo costo.

The aim of this review was to describe in a simple way the main characteristics of platelet rich Fibrin (PRF), its composition, properties and clinical application. PRF, biomaterial autologous and platelet concentrate of second generation, is a fibrin matrix containing leukocytes, platelets and growth factors, which are necessary for the healing process, giving to this biomaterial, very useful in various areas of health, including dentistry. With this review, we conclude that FRP is a real alternative to improve healing of surgical procedures and enhance other regenerative biomaterials in several areas of dentistry, in addition to accessibility and low cost.

INVESTIGATION OF BIOFILM FORMATION IN COAGULASE-NEGATIVE STAPHYLOCOCCI ISOLATED FROM PLATELET CONCENTRATE BAGS

Platelet Concentrates (PCs) are the blood components with the highest rate of bacterial contamination, and coagulase-negative staphylococci (CoNS) are the most frequently isolated contaminants. This study investigated the biofilm formation of 16 contaminated units out of 691 PCs tested by phenotypic and genotypic methods. Adhesion in Borosilicate Tube (ABT) and Congo Red Agar (CRA) tests were used to assess the presence of biofilm. The presence of icaADC genes was assessed by means of the Polymerase Chain Reaction (PCR) technique. With Vitek(r)2, Staphylococcus haemolyticus was considered the most prevalent CoNS (31.25%). The CRA characterized 43.8% as probable biofilm producers, and for the ABT test, 37.5%. The icaADC genes were identified in seven samples by the PCR. The ABT technique showed 85.7% sensitivity and 100% specificity when compared to the reference method (PCR), and presented strong agreement (k = 0.8). This study shows that species identified as PCs contaminants are considered inhabitants of the normal skin flora and they might become important pathogens. The results also lead to the recommendation of ABT use in laboratory routine for detecting biofilm in CoNS contaminants of PCs.

Avaliação do Desempenho de Culturas Convencionais na Detecção da Contaminação Bacteriana em Concentrados Plaquetários em um Hospital Universitário do Sul do Brasil / Assessment of performance of conventional culture in the detection of bacterial contamination in platelet concentrates in a University Hospital in southern Brazil

Blood transfusion is an indispensable aid to the treatmentof cancer patients. On the other hand, it involves a seriousrisk of bacterial sepsis. Platelet concentrates (PCs) arethe blood components with the highest incidence ofbacterial contamination, being responsible for mostof the septic reactions to transfusion. In this study, weassessed various conventional culture methods for thedetection of bacterial contamination in PCs. In all, 691samples of PCs (665 random donor platelets and 26apheresis platelets) from the Blood Center of the Stateof Rio Grande do Sul (HEMORGS), were analyzed. Weemployed qualitative, quantitative and daily growthculture techniques, which revealed that 2.32% of thesamples analyzed, all from random donor platelets, werecontaminated. The qualitative methodology performedbest. This result obliges us to reinforce the importanceof performing pre-transfusion bacteriological screeningon all samples of PCs, to reduce the risk of sepsis.

As transfusões sanguíneas retratam um suporte indispensável no tratamento de pacientes com câncer. Por outro lado, representam um sério risco de sepse bacteriana. Os concentrados plaquetários (CPs) são os hemocomponentes com a mais alta frequência de contaminação bacteriana, responsáveis pela maioria das reações sépticas transfusionais. Este estudo objetivou avaliar diferentes metodologias convencionais de cultura na detecção da contaminação bacteriana em CPs. Um total de 691 amostras de CPs (665 plaquetas randômicas e 26 plaquetaféreses), provenientes do Hemocentro do Estado do Rio Grande do Sul (HEMORGS), foi analisado. Foram empregadas técnicas de cultura qualitativa, quantitativa e de crescimento diário, com as quais evidenciamos 2,32% de contaminação bacteriana nas amostras analisadas, sendo todas provenientes de coleta pelo método randômico. A metodologia qualitativa apresentou o melhor desempenho. Esse fato nos permite reforçar a importância da realização de triagem bacteriológica pré-transfusional em todas as amostras de CPs para a redução dos riscos de sepse.

Evaluation of an Automated Urine Flow Cytometer for Screening of Bacterial Contamination in Platelet Concentrates

BACKGROUND: Bacterial contamination of blood products, particularly of platelet concentrates (PCs), is a major risk factor for infections caused by blood transfusion. Various methods for the detection of bacterial contamination in PCs are available or are under investigation. We evaluated the usefulness of the Sysmex UF-1000i urine flow cytometer (Sysmex Medical Electronics Co, Japan) for screening of bacterial contamination in PCs. METHODS: The PCs were inoculated with various concentrations of bacteria (Staphylococcus aureus and Escherichia coli) and were analyzed with the urine flow cytometer for bacterial counts. All the samples were diluted with normal saline (1:10) before flow cytometric analysis in order to prevent interference by the turbidity due to platelets. RESULTS: For PCs inoculated with a high number (colony forming unit, CFU) of bacteria (105 CFU/mL), the bacterial counts were significantly higher than those for uninoculated PCs analyzed by the urine flow cytometer. However, bacterial counts for PCs inoculated with bacteria of 104 CFU/mL or less and those for uninoculated PCs were not significantly different. CONCLUSIONS: An automated urine flow cytometer evaluated in this study is easy to use, and the procedure is completed in less than 5 min. Moreover, the urine flow cytometer could detect approximately 105 CFU/mL of bacteria in PCs. Further validation studies are needed to assess the usefulness of this method for screening of bacterial contamination in PCs.

Comparison of Sensitivity of Tests for Detecting Bacterial Contamination in Platelet Concentrates / 대한수혈학회지

BACKGROUND: The demand for platelet concentrates has increased for patients with hemato-oncologic diseases as well as for patients with chronic diseases. As platelet concentrates are preserved at 22~24degrees C, the chance of bacterial contamination exposure is increased, which can cause fatal outcomes. We evaluated various methods for detecting bacterial contamination in platelet concentrates. METHODS: 0.5 MacFarland standard solutions were prepared using the Staphylococcus aureus ATCC 25923 & Escherichia coli ATCC25922 strains. The platelet concentrates were inoculated with various concentrations (10(1)~10(5) CFU/mL) of bacteria and then gram staining, plate culture, broth culture and 16s RNA were used to detect bacteria. RESULTS: The gram stain method was unable to detect bacteria concentrations less than 10(4) CFU/mL. The plate culture method detected bacterial growth concentrations up to 10(3) CFU/mL, but only 1 specimen of S. aureus was detected at the lowest concentration of 10(1) CFU/mL. The broth culture method detected 10(2) CFU/mL concentrations except for samples from S. aureus and E. coli strains. Among the 10(1) CFU/mL lowest concentrations, bacterial growth detected 3 samples from S. aureus and 2 samples from E. coli. For the broth culture method, detection of bacterial growth up to 10(1) CFU/mL took 58.9 hours, it took 57.5 hours for S. aureus and E. coli respectively, and it took 43.9 hours and 49.0 hours for 10(2) CFU/mL concentrations of S. aureus and E. coli, respectively. The PCR method showed all positive results except for 1 specimen of E. coli. CONCLUSION: The broth culture method showed similar sensitivity to PCR except for the 43.9~58.9 hours of an incubation period to show positive RESULTS. Overall, the PCR method was most sensitive and rapid method for detecting bacterial contamination in platelet concentrates.

Effect of Interrupted Agitation and Removal of Leukocyte on Platelet Quality during the Storage of Platelet Concentrates / 대한진단검사의학회지

BACKGROUND: This study aimed to analyze the influence of the interruption of agitation and removal of leukocytes on platelet concentrates (PCs), and determine the maximum amount of time the agitation could be interrupted without impairing PCs' effectiveness during the storage period. METHODS: Four ABO-identical random donor platelets agitated for 24 hr were pooled, and divided into 4 units, and 2 units of them were leukoreduced. Then 52 pooled units were categorized into 4 groups, non-leukoreduced continuous agitation (Non-LRCA), non-leukoreduced interrupted agitation (Non-LRIA), leukoreduced continuous agitation (LRCA), and leukoreduced interrupted agitation (LRIA), and preserved for 6 days (total 7 days). Mean platelet volume (MPV), pH, HCO3-, pO2, pCO2, CD62P, CD61, glucose, lactate, ammonia and free fatty acid were measured during the period. RESULTS: Starting from the Day 4, the pH and HCO3- of Non-LRIA group begun to decrease while the amount of lactate production, glucose consumption, and MPV increased compared to the Non- LRCA group (P<0.01). An increase in pO2 level was observed in the interrupted agitation groups as the storage period prolonged (P<0.01). The pH levels of all the units in the agitation groups remained higher than 6.4 up to Day 7, while those of the non-leukoreduction group did so only up to Day 2, but those of leukoreduction in the interrupted agitation groups did so up to Day 4. CONCLUSIONS: The interruption of agitation reduced the platelet's capacity to utilize oxygen, increasing lactate amount and reducing pH level. However, the in vitro parameters of the Non-LRIA and Non-LRCA groups on Day 2 were similar to each other and the pH level remained at 6.4 or higher, making one day of agitation interruption possible after 24 hr of agitation. With leukocytes removed, the effective agitation interruption period may become longer.

Preparation of leukoreduced platelet concentrates from pooled buffy coats in a mixture of solutions for medical use / 中国输血杂志

Objective To develop a method to produce leukoreduced platelet concentrates(LR-PCs) from pooled buffy coats in additive solution(a mixture of solutions for medical use).Methods LR-PCs were made from 6 pooled buffy coats(12 whole-blood units) and 220g additive solution,including 90% multiple electrolytes injection solution,8% ACD-A and 2% 50g/L NaHCO3 injection solution.After centrifugation,the PCs were leukoreduced with a filter and stored in a 600ml platelet storage bag.LR-PCs were prepared in a closed system.Results Routinely produced LR-PCs(n=30) contained(2.96?0.31)?1011 platelets with a volume of(270? 32)ml.The WBC and RBC count were(1.3?0.2)?106 and(5.8?1.1)?109 per unit,respectively.The CD62P expression was(22.5? 10.6)%.After 8-day storage,the in vitro quality of LR-PCs,including pH,hypotonic shock response(HSR) and CD62P expression were 7.14?0.04,(54.0?8.2)% and(45.7?13.8)%,respectively.Conclusion In terms of the in vitro quality of LR-PCs,the method of preparing LR-PCs from pooled buffy coats in mixing solutions is feasible.

Sterilization by ~(137)Cs Gamma-ray Irradiation on Cytokine Levels in Manually Manipulated Platelet Concentrates / 中华医院感染学杂志

OBJECTIVE To study the sterilization by gamma radiation on the cytokine levels in manually manipulated platelet concentrates, and its practicability. METHODS Manually manipulated platelet concentrates were irradiated by gamma ray at 25 Gy. Detected the quantity of cytokine levels after irradiation on dd 0, 1, 3, and 5. Platelet counts and pH value were detected on the d0 and d5. RESULTS The contents of cytokines increased with storage time both in irradiation and control groups. But the contents of cytokines in the control group increased more significantly than in the irradiation one (P