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.

Comparison of the Bact/Alert blood culture system and manual culture method for detection of aerobic and facultative anaerobic bacterial contamination in platelet concentrates

For reducing bacterial contamination of platelets in the medium, PDA has approved the Bact/Alert for screening the platelet units. This study attempts to compare the Bact/Alert system and the manual culture method as far as the length of time in hours of detection is concerned. In this interventional and diagnostic study, 15 platelet units were selected randomly among 1332 units and inoculated with 10 CFU/ml of various bacteria including Streptococci, Serratia marcescens, Enterobacter cloacae, Corynebacterium diphteroid, Staphylococcus aureus and Staphylococcus epidermidis which normally contaminate platelet units. These units together with other platelet units in a blind way were tested by both Bact/Alert system and the manual method. Regarding the short expiration time of platelet units, if the length of time in hours in detection is considered as a basis for comparison, the Bact/Alert system is significantly superior to the manual method. The medium length of time in hours for detecting the aerobic bacteria by Bact/Alert system is 31 +/- 8 hours, compared with the manual method which is 61 +/- 11 hours. This shows that Bact/Alert system is nearly 2 times faster than the manual method. Bact/Alert culture system compared with the manual method is more rapid and accurate for detection of bacterial contamination thereby improving platelet safety. Regarding serious effects of these contaminations on platelet recipients, it is also necessary to try to reduce them by using GMP

Contaminação bacteriana em concentrados plaquetários: identificação, perfil de sensibilidade aos antimicrobianos e sepse associada à transfusão / Bacterial contamination on platelet concentrates: identification, antimicrobial susceptibility profile and transfusion-related sepsis

INTRODUÇÃO: Devido à sepse bacteriana associada à transfusão de concentrados plaquetários (CPs) ter sérias consequências clínicas para os pacientes, alguns procedimentos têm sido incorporados na preparação e no controle de qualidade dos componentes sanguíneos para reduzir o risco da contaminação bacteriana. Este artigo descreve a prevalência da contaminação bacteriana dos CPs que foram transfundidos, o espectro bacteriano detectado com seu perfil de sensibilidade aos antimicrobianos e as reações transfusionais nos receptores. MÉTODOS: Um total de 292 CPs (278 randômicos e 14 por aférese), proveniente do Hemocentro do Estado do Rio Grande do Sul (HEMORGS) de Santa Maria foi testado. As quantidades de 100μL e 200μL foram coletadas da porção tubular da bolsa de plaquetas e semeadas utilizando dois tipos de metodologias. RESULTADOS: Em cinco unidades(1,7 por cento; 5/292) foram isoladas bactérias pela metodologia qualitativa e apenas uma pela quantitativa. Staphylococcus epidermidis foi o microrganismo identificado em todas as amostras. Dois pacientes apresentaram sepse associada à transfusão com desfecho fatal. CONCLUSÕES: A contaminação bacteriana pelas transfusões de CPs constitui-se num importante problema de saúde pública devido a sua associação com altas taxas de morbidade e mortalidade. Neste estudo, somente microrganismos gram-positivos foram isolados sendo que nenhuma amostra obtida por aférese apresentou contaminação.

INTRODUCTION: Bacterial sepsis associated with the transfusion of platelet concentrates (PCs) results in serious clinical implications for patients. Given these implications, certain procedures have been integrated into the preparation and quality control of blood components to reduce the risk of bacterial contamination. This article describes the prevalence of bacterial contamination on transfused PCs, the bacterial spectrum detected and their antimicrobial susceptibility profile and transfusion reactions in receptors. METHODS: A total of 292 PCs (278 random and 14 per apheresis) from the Blood Center of the State of Rio Grande do Sul (HEMORGS), located in the city of Santa Maria, were tested. Quantities of 100μL and 200μL were collected from platelet bag tubing and seeded using two methodologies. RESULTS: Using the qualitative methodology, bacteria were isolated in five units (1.7 percent; 5/292), while only one was isolated using the quantitative methodology. Staphylococcus epidermidis was the microorganism identified in all samples. Two patients died of transfusion-related sepsis. CONCLUSIONS: Bacterial contamination due to PC transfusion is considered a major public health problem due to its association with high rates of morbidity and mortality. In this study only gram-positive microorganisms were isolated and none of the samples obtained by apheresis presented contamination.

Métodos de inactivación de patógenos en Plasma Fresco y Concentrados de Plaquetas / Pathogen Inactivation technology in fresh plasma and platelets concentrates

Las Tecnologías de Inactivación de Patógenos (TIPS) proveen un camino adicional para proteger el abastecimiento de sangre de agentes conocidos, emergentes y aún desconocidos. El principio precautorio refrendado por la FDA luego de la crisis del VIH/SIDA, establece que para situaciones de incertidumbre científica debiera tenerse en cuenta la posibilidad de riesgo aún en ausencia de pruebas de lo contrario. Pudiera argumentarse entonces que las TIPS representan la quintaesencia del principio precautorio. Si bien se ha acortado el intervalo entre el reconocimiento de un agente a la implementación de medidas para prevenirlo, hay a menudo un periodo de retardo temprano, durante el que una enfermedad no parece representar un riesgo para el receptor o la salud humana en general. El daño causado por ellos se ha mencionado como "una propiedad fija e inevitable de la práctica transfusional". Las TIPS para plasma fresco incluyen las siguientes: MBLT (Tratamiento con Luz y Azul de Metileno). PLT (Tratamiento con Psoralenos y Luz). RFLT (tratamiento con Riboflavina y Luz) además existe un método de Solvente Detergente para pools de plasma. Mientras que para los concentrados de plaquetas se pueden usar el Tratamiento con Luz Ultravioleta y Amotosaleno-UVA. Existe un riesgo potencial en retrasar la implementación de las TIPS mientras se aguarda la evidencia y el sistema absolutamente perfecto de ponerlas en práctica. Llegó el momento de actuar.

Pathogen Inactivation Technology (PIT) provides an additional way to protect the blood supply from agents known, emerging and yet unidentified. The precautionary principie which was first endorsed by FDA after the crisis of HIV / AIDS, states that for situations of scientific uncertainty should be taken into account the possibility of risk even in the absence of proof to the contrary. It could be argued then, that PITs represent the quintessence of the precautionary principie. Almost all of the potential for TTD is eradicated, often before the responsible agent is even recognized. Although the interval between the recognition of an agent to implement measures to prevent it has been shortened, there is often an earlier period of delay, during which a disease does not appear to represent a risk to the recipient or to human health in general. The damage caused bythem has been called a "fixed and inevitable property of transfusion practice." Techniques for pathogen inactivation of plasma include the following. MBLT (Methylene Blue and Light Treatment). PLT (Psoralens and Light Treatment). RFLT (Riboflavin and Light Treatment) in addition, there is a method of Solvent Detergent (SD) for pools of plasma. Techniques for pathogen inactivation of platelets include the following Ultra Violet Light Treatment and Amotosalen- UVA.There is great potential risk in delaying the implementation of Pathogen Inactivation Technologies (PITs) while awaiting the evidence and the absolute pedect system to put it into operation. It is time to act.

Rapid detection of salmonella typhi by culture of the mononuclear cell-platelet fraction of blood

To study the value of utilizing a new and rapid method for detecting Salmonella typhi in adult patients using the mononuclear cell-platelet fraction of blood and to compare it with the traditional methods used for the same purpose. Materials and Various specimens [blood, bone marrow and rectal swabs] were collected and cultured on standard media from 36 Jordanian patients with suspected typhoid fever at two major Jordanian hospitals, in the period from July 1992 through July 1993. Cultures using the mononuclear cell-platelet layer were performed on the blood specimens taken from all patients. Blood cultures using the mononuclear cell-platelet layer method were positive in 20 [56%] patients, and the colonies were identified [using a coagglutination technique] within 18 hours of plating. In contrast, Salmonella typhi was detected in only 15 [41%] patients using the conventional blood culture method, that required at least 3 days for identification. This study indicates that the combination of mononuclear cell-platelet layer culture and coagglutination can provide the clinician with a diagnosis of typhoid fever within a day of specimen acquisition, with a marked improvement over conventional blood culture in both time and sensitivity