Značaj direktnog testa utroška antihumanog globulina u imunohematologiji

Abstract

UVOD: Citopenija je jedna od glavnih karakteristika mnogih hematoloških bolesti. U rutinskoj transfuziološkoj upotrebi su metode detekcije prisustva antitela u serumu ili na eritrocitima bolesnika. Primena direktnog testa utroška antihumanog globulina predstavlja efikasan način da se stekne kompletan uvid u imunološka zbivanja na svim krvnim lozama, prati dinamika razvoja antitela i toka bolesti. MATERIJAL I METODE: Svim pacijentima su se iz uzoraka periferne krvi vršile sledeće analize krvi: 1) direktni antiglobulinski test mikrometodom aglutinacije u gel karticama (LISS)/ Coombs ID. Dobijeni rezultat aglutinacije mikrometodom na gelu moţe biti negativan ili pozitivan i 2) direktni test utroška antihumanog globulina metodom aglutinacije u epruveti. Očitavanje se vršilo određivanjem razlike u titru antihumanog globuli na i očitavanjem postojeće reakcije aglutinacije dobijene u uzorcima pacijenta u odnosu na rezultate reakcije aglutinacije dobijene sa uzorcima zdrave kontrolne osobe. Test se smatra o pozitivnim ukoliko se dobijala razlika u titru AHG - a za bar dva razređen ja sa pacijentovim ćelijama u odnosu na ćelije zdrave kontrolne osobe. Statistička značajnost je analizirana t - testom, Spirmanovim koeficijentom korelacije REZULTATI: Analizirano je 100 pacijenata sa dijagnozom anemije, leukopenije, limfoproliferativnih bolesti, trombocitopenije, trombotične trombocitopenijske purpure, idiopatske trombocitopenične purpure, mijelodisplastičnog sindroma, miastenije gravis i sistemskog eritematoznog lupusa pre i nakon primljene terapije. Direktni antiglobulinski test je biopozitivan u 20% slučajeva dok je direktni test utroška antihumanog globulina bio u 51%, odnosno za 31% više. Nakon primljene terapije direktni antiglobulinski test je ostao pozitivan u 18% slučajeva a direktni test utroška antihumanog globulina u 46% što je za 28% više. Utvrđivanjem povezanosti između citopenije i stepena utroška antihumanog globulina dokazano je da svi praćeni parametri utiču na stepen utroška AHG-a: hemoglobin (β=-0,579, p=0,000), hematokrit (β=-0,568, p=0,000), eritrociti (β=-0,519, p=0,000), trombociti (β=-0,617, p=0,000) i leukociti(β=-0,119, p=0,237). Takođe je dokazano da što su vrednosti posmatranih parametara veće, razlika u titru direktnog testa utroška antihumanog globulina je manja što bi išlo u prilog boljoj prognozi posmatranog oboljenja. ZAKLJUČAK: Direktni test utroška antihumanog globulina je značajno osetljiviji test u odnosu na direktni antihumani globulinski test. Postoji pozitivna korelacija između citopenije i stepena konzumacije antihumanog globulina. Smanjenje titra antitela direktnog testa utroška antihumanog globulina je jedan od pokazatelja bolje prognoze bolesti.

INTRODUCTION: Cytopenia is one of the main characteristics of many hematologic diseases. In routine use are methods of detecting the presence of antibodies in the serum or on red blood cells of patients. The application of direct consumption test of antihuman globulin is an efficient way to gain complete insight into the immunological events at all bloodlines, monitor the dynamics of the development of antibodies and disease progression. MATERIALS AND METHODS: All patients samples were tested for: 1) direct antiglobulin test by micro agglutination method in the gel card (LISS) / Coombs ID. The result obtained by micro agglutination gel can be negative or positive, 2) direct consumption test of antihuman globulin in a test tube. Interpretation is performed by determining differences in titer of antihuman globulin by reading existing reactions of agglutination in samples of the patient and compare it to the results obtained with the samples of the healthy control persons. The test is considered positive if the difference in titres obtained AHG differs for at least two dilutions of a patient's cells compared to cells of healthy control persons. Statistical significance was analyzed by t-test, Spearman correlation coefficient. RESULTS: A total of 100 patients diagnosed withanemia, leukopenia, lymphoproliferative disease, thrombocytopenia, thrombotic thrombocytopenic purpura, idiopathic thrombocytopenic purpura, myelodysplastic syndrome, myasthenia gravis and systemic lupus erythematosus were analyzed before and after receiving treatment. Direct antiglobulin test was positive in 20% cases, while the direct consumption test of anti-human globulin was 51%, that is the difference of 31%. After treatment direct antiglobulin test remained positive in 18% of cases and direct consumption test of antihuman globulin was in 46%, which is 28% higher. Determining the relationship between the degree of cytopenia and consumption of anti-human globulin showed that all monitored parameters affect the level of consumption: hemoglobin (β = -0.579, p = 0.000), hematocrit (β = -0.568, p = 0.000), erythrocytes (β = -0.519 , p = 0.000), platelets (β = -0.617, p = 0.000) and leukocytes (β = -0.119, p = 0.237). It was also proved that if the values of observed parameters are higher, difference in titer of direct consumption test of antihuman globulin is lower, which can indicate better prognosis of disease. CONCLUSION: Direct consumption test of antihuman globulin was significantly more sensitive test than the direct anti-human globulin test. There is a positive correlation between the degree of cytopenia and consumption of anti-human globulin. Decrease in antibody titer in direct consumption test of antihuman globulinis an indicator of a better prognosis of the disease.