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PubMed1953

Experimental erythroblastosis fetalis in rabbits. I. Characterization of a pair of allelic blood group factors and their specific immune isoantibodies.

A pair of blood group factors, designated G and g, was identified in rabbits by serological means. These factors were found to be alleles, and one or the other or both of them were regularly present in the red cells of every one of a large number of mongrel and inbred rabbits. The factors were not demonstrable in other tissue cells or in the body fluids. They were capable of stimulating the formation of specific immune isoantibodies when repeatedly injected into appropriate rabbits. In most instance both agglutinating and coating antibodies to the G or g factors were present in a given antiserum. The coating antibodies-which did not act as true blocking antibodies-were detected by means of an antiglobulin test (Coombs test), or by means of specific cells modified by the action of trypsin. The antibodies were heat-stable and were active over a wide temperature range; and under suitable conditions, they proved to be potent hemolysins and also capable of fixing complement. The characteristics of the antigens and antibodies of the rabbit G-g system bear a striking resemblance to those of the Rh-Hr system of man.

Animals↗
PubMed1990

Investigation into factors influencing performance of the canine antiglobulin test.

Antiglobulin (Coombs') reagents were assessed for their ability to detect immunoglobulin and complement attached to red cells. Polyspecific and monospecific reagents were prepared using a number of immunisation protocols. Performance of these antisera against control red cells was compared, in a direct Coombs' test, with samples from cases of canine autoimmune haemolytic anaemia (AIHA). A combined reagent containing two monospecific antisera (anti-IgG + anti-C3) gave optimum results. Positive control red cells were required to standardise canine Coombs' reagents for the laboratory diagnosis of AIHA. The optimum incubation temperature for the canine Coombs' test was shown to be 37 degrees C.

Anemia, Hemolytic, Autoimmune↗
PubMed2003

Immune hemolysis-serological and clinical aspects.

The differential diagnosis of anemia must consider immune hemolytic anemias as a frequent cause. Whereas detection of anti-red blood cell (RBC) alloantibodies frequently induced by immunogenic stimuli (transfusion, pregnancy) is performed by routine serology, diagnosing autoimmune hemolytic anemias or drug-induced hemolytic anemias remains a challenge, usually requiring close collaboration of a number of disciplines. Positive direct antiglobulin test (Coombs' test) represents a central criterion in diagnosing immune hemolytic anemias, leading to further detailed analyses. The most-severe type of immune-mediated hemolysis is acute intravascular hemolysis after ABO incompatible RBC transfusion. This review highlights underlying biochemical aspects, immunohematological diagnostics, and the clinical relevance of RBC allo- and autoantibodies, including paroxysmal nocturnal hemoglobinemia and drug-induced hemolysis. Finally, current and partly experimental therapeutic strategies of immune hemolytic anemias are summarized.

ABO Blood-Group System↗
PubMed1992

High-dose intravenous immune globulin therapy for hyperbilirubinemia caused by Rh hemolytic disease.

We conducted a multicenter controlled trial to test the hypothesis that high-dose intravenous immune globulin (HDivIG) therapy can modulate bilirubin production and reduce the frequency of exchange transfusions in newborn infants with Rh hemolytic disease. Thirty-four patients with Rh incompatibility proved by positive direct antiglobulin test (Coombs test) results were randomly assigned to receive conventional treatment including phototherapy, with or without additional HDivIG therapy at 500 mg/kg given for a 2-hour period as soon as the diagnosis was established. Exchange transfusions were performed if serum bilirubin concentrations exceeded the modified curves of Polácek by more than 2 mg/dl. Two patients were excluded because of protocol violations. The results in 32 infants were analyzed. In the HDivIG group, 2 (12.5%) of 16 children required exchange transfusions, whereas it became necessary in 11 (69%) of 16 children in the control group (p less than 0.005). Bilirubin levels in the HDivIG group were lower despite reduced frequency of exchange transfusions. No side effects of HDivIG treatment were observed. We conclude that HDivIG therapy by a yet unknown mechanism reduces serum bilirubin levels and the need for blood exchange transfusions in children with Rh hemolytic disease.

Anemia↗
PubMed2001

An unusual increase of blood anti-beta 2-glycoprotein-I antibody but not antiphospholipid antibody in cerebral ischemia--a case report.

An abnormal increase in anti-beta2-glycoprotein I antibodies (abeta2GPI) is capable of producing thrombosis and the vasculopathy-simulating antiphospholipid antibody (aPL). However, it is rarely described in cerebral ischemia without an association with aPL. The authors report a middle-aged man who experienced recurrent cerebral ischemia and diffuse cerebral stenosis without the apparent traditional cardiovascular risk factor. He was free of antiphospholipid/cofactor syndrome (APCS) and systemic lupus erythematosus (SLE). An increase of blood abeta2GPI was detected in serial measurements. The aPL, Venereal Disease Research Laboratory (VDRL) test, Coombs' test, and antinuclear factor were negative. Activated partial thromboplastin time was normal. This patient is a reminder to consider abeta2GPI in an unexplained recurrent cerebral thrombosis and cerebral artery stenosis even when the typical clinical manifestation or laboratory data of APCS is absent.

Antibodies, Antiphospholipid↗
PubMed1991

[The use of the latex agglutination reaction for the diagnosis of a Brucella infection].

The authors have developed the optimum conditions for the preparation of antigenic diagnosticum based on latex manufactured in the USSR. To sensitize latex with the diameter of microspheres equal to 0.83 microns, Brucella polysaccharide was used in a dose of 100 micrograms/ml. As stabilizer, polyvinylpyrrolidone at a concentration of 0.1% was used. The specificity and sensitivity of the diagnosticum were studied in analysis of serum samples taken from 102 healthy donors and patients with infectious diseases of nonbrucellar etiology and from 120 patients with different forms of brucellosis. The specificity of the diagnosticum was found to be 94.1% and its sensitivity, 77.5%. Comparative study of the latex agglutination test with other serological tests showed that the former test is highly effective both in acute and chronic forms of the disease. A high degree of correlation between the agglutination test, Coombs' test, the passive hemagglutination test and the latex agglutination test was established (r = 0.83, 0.72 and 0.62, respectively).

Antibodies, Bacterial↗
PubMed1986

Cytochemical and cellbiological investigations of the signal function of the erythrocyte plasmalemma--the membrane structure as code for cell life span.

The erythrocyte membrane was characterized with regard to its alteration of different degrees. A very important process of the membrane alteration is the expression of IgG receptor sites, which is connected with the binding of autologous IgG at the membrane. Several detection methods for membrane bound IgG were described: immunocytochemical techniques (Anti IgG--Anti Ferritin-sandwich technique, Anti IgG-Protein A-Gold- and the Silver enhancement technique), Antiglobulin agglutination test (Coombs test) and erythrocyte-macrophage test. Moreover, the topo-optical toluidine blue reaction and the ultra-histochemical NAD(H) oxidase reaction were checked to characterize the erythrocyte-macrophage interaction. Finally, the microvesiculation is understood as special form of the erythrocyte membrane disintegration, which is connected with a remodelling of an intact plasmalemma. The findings presented here summarize the fact that the membrane associated IgG regulates an immune-signal for the elimination of the erythrocytes in the Reticulo-Histiocytic-System in vivo and in vitro. Therefore, the structural integrity of the erythrocyte plasmalemma is the code for the cell life span.

Animals↗
PubMed1985

Transient presence of an anti-D alloantibody.

A 37-year-old man with blood type O Rh-negative displayed a sudden, transient occurrence of an anti-D antibody shortly after receiving only type O Rh-negative packed red blood cells and Rh-positive random donor platelets. Testing of the blood products that were transfused demonstrated a high anti-D antibody titer in one of the packed red blood cell units that the patient received one day before the antibody was detected. Passively acquired antibodies should be routinely considered when evaluating the sudden occurrence of a positive indirect or direct antiglobulin test (Coombs' test) in a patient. Early recognition of this phenomenon will prevent unnecessary delays or inappropriate therapy.

Adult↗
PubMed1984

Red cell, white cell and platelet autoantibodies in visceral leishmaniasis.

The blood of twelve patients with parasitologically proved visceral leishmaniasis was examined for the presence of antibodies against blood cells. The direct antiglobulin test (Coombs' test) was positive in ten. IgG and complement were found on the red cells (alone or combined), but no IgA or IgM. In the serum of two, warm antibodies were found in the bromelin test. Pseudo-agglutination was detected in 8. Investigations for white cell antibodies included the lymphocytotoxicity test (positive in 10), the leuco-agglutination test (positive in 1) and the indirect immunofluorescence test on lymphocytes (none positive) and granulocytes (2 positive). Platelet autoantibodies were demonstrated in the direct (10 positive) and indirect (7 positive) platelet suspension immunofluorescence test. How important these various antibodies are in influencing the lifespan and functions of the respective cells and thus in the pathogenesis of the pancytopenia of kala-azar, can not be concluded from this work but the absence of a direct correlation between haematological values and presence or absence of antibodies and/or their titres argues against a primary role of these antibodies.

Adolescent↗
PubMed1984

[Anti-brucella agglutinins in aborted women].

52 (6.46 per cent) out of 805 women who aborted spontaneously in Gujarat state, India, were found to have positive Brucella agglutin. Infection with brucella was further confirmed by various serological tests (Rapid plate test, standard agglutination test, Rose Bengal test, Coombs' test, complement fixation test, indirect bacterial haemagglutination test, gel diffusion test and determination of C-reactive protein). Culturing 650 blood clots, 300 placentae and 800 samples of urine of women who aborted gave totally negative results. Women who were selected for this study immediately after admission to the Civil Hospital, Ahmedabad, India, came from rural areas and belonged to the farming community. They had a history of contact with animals and were of the lower socio-economic group. The relationship between brucellosis and spontaneous abortion is discussed. The aborted women would most probably have contracted the infection from their domesticated animals, since brucellosis is prevalent amongst the animals in the State of Gujerat.

Abortion, Habitual↗
PubMed1993

[High-dose immunoglobulin therapy of hyperbilirubinemia in rhesus incompatibility].

We conducted a multicenter controlled trial to test the hypothesis that high intravenous doses of immunoglobulin (HDivIg) can modulate the bilirubin production and reduce the frequency of exchange transfusions in newborn infants with rhesus incompatibility. Thirty-four patients with rhesus incompatibility proven by positive direct antiglobulin test (Coombs test) were randomly assigned to conventional treatment including phototherapy, with or without additional HDivIg at 500 mg/kg given over 2 h, as soon as the diagnosis was established. Exchange transfusions were performed if serum bilirubin concentrations exceeded the modified curves of Polácek by more than 2 mg/dl. The results in 32 infants were analyzed. In the HdivIg-treated group, 2 of 16 (12.5%) children required exchange transfusions, whereas it became necessary in 11 of 16 (69%) in the control group (p < 0.005). Bilirubin levels in the HDivIg-treated group were lower despite a reduced frequency of exchange transfusions. We conclude that HDivIg by a yet unknown mechanism reduces bilirubin serum levels in children with rhesus incompatibility and the need for exchange transfusions.

Bilirubin↗
PubMed2004

Laboratory diagnosis of brucellosis in a rural endemic area in northeastern Spain.

Sera obtained from 62 patients from four mountain counties in Catalonia (Northeastern Spain), in whom brucellosis had been diagnosed on the basis of clinical evidence and/or personal history, were analyzed using the rose Bengal test, standard serum agglutination test (SAT), Coombs' test, ELISA, and complement fixation. The diagnosis was further confirmed through blood cultures. Clinical evidence, epidemiology, and the results from serologic tests were used to assign patients to one of two groups: group 1 (n = 38) patients had primary infections, whereas group 2 (n = 24) patients had been previously exposed to the microorganism, i.e. re-infection of group 2 individuals occurred after long periods of time during which no active infection by Brucella had been detected. Receiving-operating charts (ROC) were used to determine the diagnostic value of the different tests and to establish discriminant values. Blood culture was a valuable diagnostic tool in group 1 (0.92 sensitivity) but was inappropriate in group 2 (0.08). The combination of positive rose Bengal test and agglutination >/=1/160 was valid for diagnosis in group 1. In group 2, agglutination <1/160 (including negative agglutination) did not rule out brucellosis. The combination of positive rose Bengal test and Coombs' test >/=1/320 was the best diagnostic criterion (0.8 specificity; 1 sensitivity). ELISA (for IgG, IgM, or both) did not improve diagnostic accuracy.

Brucellosis↗
PubMed1985

Autoimmune hemolytic anemia as the presenting manifestation of malignant thymoma.

Severe autoimmune hemolytic anemia is described as the presenting manifestation of malignant thymoma. The hemoglobin level was 5.7 g/dl, the direct IgG antiglobulin test (direct Coombs' test) was strongly positive and the indirect Coombs' test was weakly positive. Prompt remission of the hemolytic process was achieved by thymectomy combined with corticosteroid therapy; the hemoglobin level rose to 12.2 g/dl and both the direct and indirect Coombs' tests became negative. To our knowledge, this is the first case to be described in which autoimmune hemolytic anemia was the presenting manifestation of malignant thymoma. Autoimmune hemolytic anemia may be considered as a manifestation associated with malignant thymoma.

Anemia, Hemolytic, Autoimmune↗
PubMed2004

Evaluation of Brucellacapt for the diagnosis of human brucellosis.

OBJECTIVE: To evaluate the role of Brucellacapt in the diagnosis of human brucellosis, and the correlation with the evolution of the disease. METHODS: Twenty-six patients who were admitted to the General Hospital of Albacete (Spain) over a 2-year period and diagnosed with brucellosis were included in the study. One hundred and twenty-three serum samples collected at the time of diagnosis and at intervals during and after treatment were tested by the Coombs test, the standard seroagglutination test (SAT), and Brucellacapt (a new test based on an immunocapture-agglutination technique). To study the specificity of Brucellacapt, sera from 20 patients with other infectious diseases and 20 sera from healthy donors were included in this study. RESULTS: The sensitivity of the Brucellacapt at the moment of diagnosis was similar to the Coombs test (96 and 100%, respectively), somewhat higher than that of SAT (73%). And the specificity of the Brucellacapt (97.5%) was less than SAT and the Coombs test, that was 100%. The correlation between the classical serological tests and Brucellacapt, showed that titers in Brucellacapt and Coombs test of patients were both similar in a range of 1-2 dilutions. The correlation between Brucellacapt and Coombs (r=0.14) and between Brucellacapt and SAT (r=0.0) did not reach statistical significance. However, the correlation coefficient between Coombs and SAT was r=0.8. CONCLUSIONS: Brucellacapt and Coombs tests showed a similar sensitivity and specificity in the diagnosis of human brucellosis. In addition, as Coombs test, Brucellacapt could help to diagnose patients with long evolution of brucellosis that are not detected with SAT.

Adolescent↗
PubMed2003

Clinical utility of a dipstick assay in patients with brucellosis: correlation with the period of evolution of the disease.

To examine the clinical utility of a dipstick assay for the detection of Brucella-specific IgM antibodies, and the correlation with the evolution of the disease. Twenty-six patients who were admitted to the General Hospital of Albacete (Spain) over a 2-year period and diagnosed with brucellosis were included in the study. One hundred and twenty-five serum samples collected at the time of diagnosis and at intervals during and after treatment were tested by the Coombs test, the standard seroagglutination test (SAT), the SAT in the presence of dithiothreitol (DTT-SAT), and a dipstick assay for the detection of Brucella-specific immunoglobulin M (IgM) antibodies. The sensitivity of the dipstick assay at the moment of the diagnosis was similar to that of the SAT (62% and 73%, respectively), somewhat higher than that of the DTT-SAT (50%), and lower than that of the Coombs test (100%). Patients with a negative dipstick test at the moment of diagnosis displayed a period of evolution of the disease longer than that of the dipstick-positive patients. After the beginning of therapy, the detection rate of the dipstick assay decreased faster than those of the SAT, the DTT-SAT, and the Coombs test. Thirty days after the start of therapy, the detection rate of the dipstick assay had decreased to 7%, whereas that of the SAT and DTT-SAT was 46%, and that of the Coombs test was still 92%. The dipstick assay could be used as a rapid diagnostic test for patients in the early stages of illness. Patients with a long period of illness will probably have a negative dipstick test, and could be diagnosed with the aid of the Coombs test and classical clinical findings.

Agglutination Tests↗
PubMed1997

Serological characteristics of systemic lupus erythematosus from a hospital-based rheumatology clinic in Kuwait.

Thirty-one consecutive patients with SLE were screened for antinuclear antibody (ANA), anti-DNA antibodies, extractable nuclear antigen antibodies (anti-ENAs) including anti-Sm, anti-RNP, anti-SSA (anti-Ro), anti-SSB; (anti-La), anti-Scl-70, rheumatoid factor (RF), C-reactive protein (CRP), C3 and C4 levels, anti-cardiolipin antibodies (aCL), biologically false positive serological test for syphilis (BF-STS) using VDRL test and Coombs' test. The age of the patients ranged from 11 to 52 year with a median of 29 year; female to male ratio of 5:1. There were 21 Kuwaitis, four Egyptians, three from the Indian subcontinent, two Filipinos and one Syrian. Main clinical categories of SLE were: mild cutaneous SLE in 12 (38.7%), clinical antiphospholipid syndrome (APS) secondary to SLE in 8 (25.8%), haematological manifestations of SLE in 5 (16.1%), renal lupus in four (12.9%), neuropsychiatric in three (9.7%), others (6.4%). Clinical features overlapped in several patients. ANA was positive in 96.8% (mean value 891.61 units/ml), anti-DNA in 35.5% (mean value 56.4 units) that was lower than expected and could be due to selection bias as the patients were from a rheumatology clinic, anti-ENA in 42%, anti-Sm 13% that was lower than other non-Caucasian populations, anti-RNP 13%, anti-SS-A in 35.5%, anti-SS-B in 19.4%, Scl-70 in 13%, CRP in 71% (moderate 58%, very high 13%); C3 mean 1.52 mg/ml (3.2% low levels), C4 mean 0.35 mg/ml (32% low levels), anticardiolipin mean GPL 35.35 units (high 58%), mean MPL 10.61 units (high 26%), BF-STS in 6%, Coombs' test in 6%, RF positive in 36%. The only significant positive clinical associations observed were those of renal involvement with anti-DNA antibodies (P = 0.042), and clinical antiphospholipid antibody syndrome with aCL antibodies (P = < 0.05).

Adolescent↗
PubMed1990

Use of delayed-type hypersensitivity test to diagnose brucellosis in calves born to infected dams.

Calves (n = 2) born to dams with experimentally induced brucellosis, and calves (n = 4) born to dams with naturally occurring infection were examined by the delayed-type hypersensitivity (DTH) test for possible B. abortus infection. The results were compared with the serum agglutination test, complement fixation test, and Coombs test. Five calves were nursed by their dams for 8-10 weeks after birth. One calf was separated from its dam and fed artificial milk. Three to five months after birth, four calves tested seropositive in the serologic tests. Antibodies were detected in one calf as early as 1 week after birth. The calf fed on artificial milk was seronegative 4-5 weeks after birth. All calves reacted to the DTH test antigen from week 12 until the end of the experiment, even though serologic tests were negative. We conclude that the DTH test is a valuable technique for diagnosing Brucella in calves born to infected dams.

Agglutination Tests↗