Avaliação da validação da pesquisa de anticorpos irregulares no Hemocentro Regional de Montes Claros / Validation assessment of irregular antibodies investigation at the Regional Blood Center in Montes Claros City / Evaluación de la validación de la búsqueda de anticuerpos irregulares em la Región Hemocentro de Montes Claros

Objetivo: Analizar lavalidación de la detección de anticuerpos irregulares (PAI) mediante el uso del reactivo de control de Coombs en muestras de sangre tomadas de Febrero 2015 a Agosto 2016. Métodos: Estudio de naturaleza observacional, retrospectivo y prospectivo, con los procedimientos técnicos de carácter documental, que se sucederá em Laboratorio de Inmunohematología del Hemocentro Regional de Montes Claros - MG. Resultados: Se observó durante la encuesta que después de la no validación de algunos testes y sucedida la repetición del mismos individuos, no se ha encontrado la validación, por lo tanto requeiendo otra repetición hasta que la validación de la muestra. Esto plantea la posibilidad de interferencia que no sea el conocido y discutido, ya que la repetición se realiza aisladamente el análisis crítico de todos los pasos del proceso. Conclusión: El bajo porcentaje de resultados no validados ratifica la prueba de validación antiglobulínico es un buen método para confirmar el resultado de la búsqueda de anticuerpos irregulares

Objetivo: Analisar a validação da pesquisa de anticorpos irregulares (PAI) através da utilização do reagente Controle de Coombs em amostras sanguíneas coletadas de Fevereiro de 2015 à Agosto de 2016. Métodos: Estudo de natureza observacional, retrospectiva e prospectiva, apresentando procedimentos técnicos de caráter documental, a ser realizado no Laboratório de Imunohematologia do Hemocentro Regional de Montes Claros - MG. Resultados: Foi observado durante a pesquisa que após a não validação de alguns testes e realizada a repetição dos mesmos isoladamente, não foi constatado a validação sendo necessário outra repetição até que essa amostra validasse. Esse fato levanta a possibilidade de outras interferências além das conhecidas e discutidas, uma vez que a repetição foi realizada isoladamente analisando criticamente todas as etapas do processo. Conclusão: O baixo percentual de resultados não validados ratifica que o teste de validação antiglobulínico é um bom método para confirmar o resultado da pesquisa de anticorpos irregulares

Objective: The study's purpose has been to assess the validation of irregular antibodies investigation using the Coombs control reagent in blood samples collected over the period from February 2015 to August 2016. Methods: It is a observational, retrospective and prospective study, which presents technical procedures bearing a documentary character, and that was performed at the Laboratory of Immunohematology from the Regional Blood Center in Montes Claros-MG. Results: During the research, it was observed that after the non-validation of some tests and its repetition was then performed alone; the validation was not verified and once again a repetition was necessary until this sample was defined as validated. This fact raises the possibility of other interferences beyond those both known and discussed; bearing in mind that the repetition was carried out in isolation and also all stages of the process were performed under scrutiny. Conclusion: The low percentage of non-validated results ratifies that the antiglobulin validation test is a good method to confirm the result of the search for irregular antibodies

Caracterización de los antígenos y anticuerpos eritrocitarios en pacientes en espera de trasplante renal / Red blood cell antigens and antibodies in patients awaiting renal transplantation

Introducción: la importancia de los grupos sanguíneos para la terapia transfusional y el trasplante es bien conocida. La presencia de anticuerpos eritrocitarios puede mediar reacciones transfusionales hemolíticas severas y rechazo de trasplante. Objetivo: caracterizar los antígenos y anticuerpos eritrocitarios en pacientes en espera de trasplante renal.Métodos: se realizó un estudio prospectivo en 980 pacientes en espera de trasplante renal considerados aptos para trasplante, en el periodo comprendido entre julio de 2013 y julio de 2014. Se investigó la frecuencia de los grupos sanguíneos ABO, Rh (DCcEe), Kell (K), Duffy, Kidd y Lewis y se realizó la pesquisa de auto y de aloanticuerpos eritrocitarios a través de las prueba de antiglobulina directa e indirecta (Coombs) y la técnica de polietilenglicol. Resultados : el grupo sanguineo 0 fue el más frecuente, seguido del A, el B y el AB. Dentro de los fenotipos RhD positivos, el DCCee predominó en los individuos blancos y el Dccee en los no blancos. El RhD negativo (ccee) fue más frecuente en blancos que en no blancos. La distribucion del antigeno Kell fue similar en ambos grupos. Se identificaron 14 pacientes (1,4 por ciento) con prueba de Coombs directa positiva, y aloanticuerpos eritrocitarios en 35 pacientes, para una frecuencia de aloinmunización eritrocitaria del 3,6 por ciento. Predominaron los anticuerpos anti - Rh y contra el antigeno Kell. La técnica de polietilenglicol detectó un mayor número de anticuerpos que la PAI, especialmente contra el antigeno RhD, aunque la comparación no fue estadisticamente significatica. Conclusiones: la frecuencia de aloinmunización eritrocitaria es menor que las comunicadas en otros estudios y se relacionó con los antecedentes transfusionales. Se recomienda realizar la pesquisa de auto y aloanticuerpos eritrocitarios a todos los pacientes con enfermedad renal crónica en lista de espera de trasplante(AU)

Introduction: The importance of blood groups in transfusion therapy and transplant is very well known. The presence of red blood cell antibodies can mediate severe hemolytic transfusion reactions and transplant rejection. Objective: To characterize red blood cell antigens and antibodies in patients awaiting renal transplantation. Methods: A prospective study in 980 patients in waiting list for renal transplantation in the period from July, 2013 to July, 2014 was carried out. The frequency of ABO, Rh (DCcEe), Kell (K), Duffy, Kidd and Lewis blood groups, and the screening of red blood cells auto and alloantibodies by the direct and indirect antiglobulin test (Coombs) and the polietilenglicol technique were investigated. Results: Blood group O was the most frequent followed by A, B and AB. DCCee phenotype was frequent in white individuals and Dccee in non-white. RhD negative (ccee) was more frequent in whites than in non-whites. Distribution of Kell antigen was similar in both groups. Direct antiglobulin test was positive In 14 patients (1,4 percent) and red blood cell alloantibodies were identified in 35 patients for a frequency of alloimmunization of 3,6 percent. Anti-Rh anti-K antibodies were the alloantibodies most frequently identified. The polietilenglicol technique detected a higher number of antibodies than the indirect antiglobulin test, specially against RhD antigen, although the comparison was not statistically significant. Conclusions: The frequency of alloimmunization is smaller than those communicated in other studies which was related to transfusion records. A periodic red blood cell auto and alloantibodies screening is recommended in all patients awaiting renal transplantation(AU)

Producción de suero de Coombs en carneros inmunizados con globulinas humana opsonizadas / Coombs serum production in sheep immunized with human globulin opsonized

Se obtuvo suero antiglobulínico (Coombs) con el empleo de un inóculo consistente en un inmunocomplejo (IC) inmunoglobulina (Ig) humana-antiglobulina humana en carnero, como opsonina para favorecer la respuesta inmune. Se inmunizaron 18 carneros divididos en 3 grupos de 6: el primero y el segundo destinados a producir anti-IgG y anti-C3, respectivamente. Estos, a su vez, subdivididos en subgrupo A: en el que se empleó el método tradicional de obtención de suero de Coombs; y B: en el que se usó el adyuvante completo de Freud en la dosis inicial y el IC en la fase de mantenimiento. Al tercer grupo solo se le administró el IC puro (subgrupo A) y en una dilución 1:200 (subgrupo B). En los carneros de los subgrupos 1B y 2B se obtuvieron títulos más elevados de anti-IgG y anti-C3dg, que en los inmunizados por el método tradicional. La respuesta de anticuerpos en los animales que se inmunizaron con los IC (3A y 3B), fue más rápida y de mayor título que las obtenidas por el método tradicional (1A y 2A) o el método combinado (1B y 2B). La respuesta en el subgrupo 3B fue más prolongada, al parecer por un efecto de dosis

An antiglobulin serum (Coombs) was obtained using a consistent inoculums in a immunocomplex (IC) the human immunoglobulin (Ig)/human antiglobulin in the seep by example, the opsonin to favor the immune response. Eighteen sheeps were immunized divided into three groups of 6 each: The first and second aimed to produce anti-IgG and anti-C3, respectively. In turn, these were divided into the A subgroup: in which we used the traditional method of Coombs's serum obtaining and B group in which we used the Freud's whole adjuvant in initial dose and the IC in the maintaining phase. Third group received the pure IC (A subgroup) and at a dilution of 1:200 (B subgroup). In sheeps from the 1B and 2B subgroups it was possible to obtain higher titration of anti-IgG and anti-C3dg than those immunized by means of the traditional method. The antibody response in animals immunized with the ICs (3A and 3B) was faster and of higher titration than those obtained by traditional method (1A and 2A) or the combined method (1B and 2B). The response in the 3B subgroup was lengthier apparently by a dose effect

Detection of Rh antibodies using two low ionic diluents: extension of the incubation time and the number of Rh antibodies detected.

BACKGROUND: Low ionic strength solution (LISS) is used to increase the rate of association of antibody to the corresponding antigen during antibody detection tests. A number of LISSs are available on the market. AIMS: The efficiency of two commercial low ionic diluents, DiaMed ID-CellStab and Inverclyde LISS were assessed using the DiaMed-ID LISS Coombs microtube column system and an incubation time varying from 15 to 35 min. MATERIALS AND METHODS: One-hundred samples containing five Rh antibodies (anti-D, anti-C, anti-E, anti-c and anti-e) were tested against commercial red cells using the two low ionic diluents after 15, 25 and 35 min. RESULTS: The Inverclyde LISS detected 91, 95 and 96% of the Rh antibodies compared to 78, 79 and 83% for ID-CellStab after 15, 25 and 35 min incubation time, respectively, for both methods. CONCLUSIONS: The Inverclyde LISS is a more suitable and efficient diluent than ID-CellStab for the detection of Rh antibodies. The sensitivity of Rh antibody detection increased for the both methods as the incubation time increased.

Anemia hemolítica autoinmune por anticuerpos calientes: reporte de un caso pediátrico / Autoimmune hemolytic anemia, warm antibody: a case report pediatric

Las anemias hemolíticas autoinmunitarias se caracterizan por la presencia de inmunoglobulinas en la superficie eritrocitaria dirigidas contra los determinantes antigénicos de los hematíes. La anemia hemolítica autoinmune por anticuerpos calientes se caracteriza porque los autoanticuerpos actúan a la temperatura del organismo (37°C), son de clase IgG y la hemólisis es predominantemente extravascular, siendo el tipo más frecuente de anemia hemolítica autoinmune en los niños de 2-12 años de edad. Sus manifestaciones clínicas son postración, palidez, ictericia, fiebre y hemoglobinuria. El diagnóstico de las AHAI se establece con la prueba de Coombs. La administración de corticoesteroides constituye el tratamiento inicial de elección. Se presenta el caso de una preescolar femenina de tres años de edad procedente del medio rural, quien exhibe las características clínicas, paraclínica y epidemiológicas de anemia hemolítica autoinmune por anticuerpos calientes, con respuesta satisfactoria a la terapia con esteroides.

Role of gel based technique for Coomb's test.

The direct (DAT) and indirect antiglobulin tests (IAT) are one of the most important diagnostic tools used in the investigation of immune mediated disorders. Recently, transfusion laboratories have seen the introduction of column technology in the form of the gel technology (GT). Aim of this study is to compare the conventional tube tests (CTT) and the GT for Coomb's test and to evaluate their sensitivity and specificity. 1656 samples were included in this study, in which 1054 samples were subjected to IAT and 602 samples were subjected to DAT Of the 602 samples tested for DAT, 587 (97.5%) showed concordant DAT results. DAT by the GT could detect 8.6% positivity as compared to 6.1% by CTT. The sensitivity and specificity of the GT was 100% and 97.3% respectively and its negative predictive value was 100%. Among the 1054 samples for IAT, 1041 (98. 8%) showed concordant results. The IAT by the GT showed 6.6% positivity as compared to 5.4% positivity by CTT The sensitivity, specificity and the positive and negative predictive value were 100%, 97.7%, 81.4% and 100% respectively. In conclusion, the GT is a better alternative to the CTT for both DAT and IAT. The GT is highly recommended to be implemented as a routine method of testing in all zonal / regional blood transfusion centers.

Comparison of standard tube and microagglutination techniques for determining brucella abortus antibodies

A microagglutination method for determining the agglutinating and blocking antibodies to Brucella abortus is described. A collection of sera from cattle in 2 rural areas of Saudi Arabia were tested by the microagglutination methods and the standard methods in tube. The results were compared and showed that where discrepancies occurred, these were due to the microagglutination methods being more sensitive. It is concluded that these are suitable methods for screening animal stocks