Hemagglutination Inhibition Method Research Articles

Immunoassay for renin: measurement of renal renin content in autopsied hypertensive patients.

Immunoassay for renin in human kidneys was undertaken by a hem-agglutination inhibition technique using high titer antirenin produced in rabbits. Human renin, highly purified by DEAE-cellulose and CM-sephadex column chromatography, was used for sensitization of sheep blood cells. Renin was extracted from kidneys of 11 normotensive and 27 hypertensive subjects through the five steps of the purification procedure described by Haas et al. The inhibition titer of antirenin was found to have a linear relationship with the concentrations of renin. However, in assay of lower concentrations of renin, such as plasma renin, there was some difficulty in reading the end-point of reactions since much diluted antirenin serum had to be used. The pressor effect of renin on unanesthetized dogs estimated by the direct method was compared with the results obtained by the hemagglutination inhibition method. Excellent agreement was obtained between results by both methods. The limits of estimation were 0.05 (Goldblatt) unit per gram of kidney tissue for dog assay and 0.001 unit per gram for the hemagglutination inhibition method. The mean value of renin for normal kidney was 0.105±0.033 U/g. The renin content was slightly higher in essential hypertension (0.222±0.066), greatly elevated in malignant hyperten-sion (0.560±0.376), and low in hypertension caused by chronic glomerulonephritis (0.054±0.020), chronic pyelonephritis (0.040), polycystic kidney (0.006), and Cushing's syndrome (0.008).

The occurrence of Ag determinants in different lipoproteins.

Abstract. The occurrence of Ag determinants on low density (LDL), very low density (VLDL) and Lp(a) lipoproteins was studied with the passive hemagglutination inhibition method. All Ag specificities found in LDL lipoproteins of a person could also be demonstrated in the plasma, VLDL and Lp(a) lipoprotein of this person. After delipidation of LDL with sodium deoxycholate, the Ag antigens a1 and g could still be demonstrated in the apoprotein while the Ag antigens x and y could not.

Automated continuous-flow hemagglutination inhibition test for hepatitis B antigen in blood.

Abstract. An automated continuous‐flow hemagglutination inhibition (HI) method for hepatitis B antigen using purified HAA‐conjugated red cells has been developed in these laboratories and has been used to test 497 blood donors. For comparison, counter electrophoresis (CEP) and microtiter complement fixation (CF) tests were done concurrently. 14 samples were found to be HAA‐positive by all three tests. One additional sample was negative by CF but HI and CEP positive. The automated HI test proved very simple to operate. The results are available in 27 min at the rate of 60 samples/h.

Studies on the secretion of blood group substances. II. Observations on the red cell phenotype Le(a-b-x-).

Abstract Salivas from 35 type Le(a‐b‐x‐) subjects were tested for Lea,Leb, Lex, A, B and H substances by both manual and automated quantitative hemagglutination inhibition methods. For comparison, similar data were gathered from 226 Lewis positive adults. Among the 13 Ma‐bx‐) nonsccretors, 7 had small quantities of salivary Lea activity and 11 had low concentrations of salivary Lex by manual methods. None showed any salivary Lebactivity. By the automated method, low concentrations of Leaactivity were demonstrable in 12. Among the 22 Ma‐b‐x‐) ‐tors, 6 had minute quantities of salivary Lewis activity, mainly Leb. The Leb activity could not represent cross reactivity with H substance because most of those subjects with the highest H activities had no Leb activity.Our quantitative studies show, when salivary Lewis activity is present in individuals whose red cells era type Ma‐bx‐), that the activity is much lower in comparison with individuals whose red cells are Lewis positive. Thus, based on quantitative salivary studies, it is possible to distinguish between Lewis positive (Len and Lewis negative (le/le) individuals.These observations suggest that the le gene is not a completely inert allele but that it governs the biosynthesis of serologically active substances. The Ze gene can be regarded as an inefficient Le which produces exactly the same products in a qualitative way as the Le gene but quantitatively far less. Another possibility is, as recently suggested, that the le gene actually governs the biosynthesis of distinct products Lec and Led and that salivary Lea and Leb activities among Lewis negative subjects represent cross reactivity with these products.

Studies on the Secretion of Blood Group Substances

Salivas from 35 type Le(a-b-x-) subjects were tested for Le^a, Le^b, Le^x, A, B and H substances by both manual and automated quantitative hemagglutination inhibition methods. For comparison, similar data were gathered from 226 Lewis positive adults. Among the 13 Le(a-b-x-) nonsecretors, 7 had small quantities of salivary Le^a activity and 11 had low concentrations of salivary Lex by manual methods. None showed any salivary Le^b activity. By the automated method, low concentrations of Le^a activity were demonstrable in 12. Among the 22 Le(a-b-x-) secretors, 6 had minute quantities of salivary Lewis activity, mainly Le^b. The Le^b activity could not represent cross reactivity with H substance because most of those subjects with the highest H activities had no Le^b activity. Our quantitative studies show, when salivary Lewis activity is present in individuals whose red cells era type Le(a-b-x-), that the activity is much lower in comparison with individuals whose red cells are Lewis positive. Thus, based on quantitative salivary studies, it is possible to distinguish between Lewis positive (Le/) and Lewis negative (le/le) individuals. These observations suggest that the le gene is not a completely inert allele but that it governs the biosynthesis of serologically active substances. The le gene can be regarded as an inefficient Le which produces exactly the same products in a qualitative way as the Le gene but quantitatively far less. Another possibility is, as recently suggested, that the le gene actually governs the biosynthesis of distinct products Le^c and Le^d and that salivary Le^a and Le^b activities among Lewis negative subjects represent cross reactivity with these products.

Virus antibody levels in systemic lupus erythematosus.

Antibody titers to a group of viral antigens have been determined in sera from patients with systemic lupus erythematosus (SLE), control groups with inflammatory diseases and normals. Mean titers in SLE sera for all viruses tested were significantly greater than in four control groups, but not greater than in active tuberculosis, both by the complement‐fixation (CF) and hemagglutination‐inhibition (HI) methods. By the CF method, only measles virus showed significantly higher titers in SLE than in all control groups; by the HI method, measles antibody titers were higher in SLE than in all groups but tuberculosis. There was no correlation between antibody titers and gammaglobulin levels. The results indicated a moderate though variable overall hypereactivity in SLE to the viral antigens tested.

Antigenic relationship between the C4 inactivating substance produced by KB cell cultures and the first component of human complement (C1).

The antigenic relationship between the C4-inactivating substance produced by KB cell cultures and the first component of human complement was studied. The antigenic identity of the two substances has been demonstrated applying Ouchterlony's double diffusion method and a special haemagglutination-inhibition method developed by the authors. The site of synthesis of human C1 and its subcomponents has been discussed.

Method for Typing Antisera to Herpesvirus hominis by Indirect Hemagglutination Inhibition

A test for typing antisera to Herpesvirus hominis that uses the method of indirect hemagglutination inhibition is described. The test, which is based upon the differential absorption of herpes antisera by preparations of type 1 and type 2 antigens, is rapidly and easily performed. The results permit some conclusions to be drawn regarding the antigenic relationships between the two virus types. Some of the practical limitations of the test are discussed.

Staphylococcal clumping on microtiter plates: a rapid, simple method for measuring fibrinogen split products.

A rapid, simple method for measuring fibrinogen split products in serum is described. It is based on clumping of staphylococci after serial dilution of test samples on a microtiter plate. The assay can be completed in less than 30 min.; its usual sensitivity is 0.75 μ g. per ml. (in fibrinogen equivalents). The sera of 52 blood bank donors had a range of 0.75 to 9 μ g. per ml. (mean ± , 2.7 ±2.1 ±g. per ml.), with only one value > 6 μ g. per ml. The results by this method were in good agreement with those obtained by a hemagglutination-inhibition method for frozen sera of patients with evidence of the intravascular coagulation and fibrinolysis (ICF) syndrome. Analysis in cases of suspected ICF should be done on fresh sera.

Gel filtration of ABH blood group substances : I. Fractionation of ABH substances of human saliva

Human saliva from secretors of groups A, B, O and AB was gel filtered on Sephadex G-200 and G-100 columns, and on thin layers and the eluates were tested for A, B and H antigens by the haemagglutination inhibition method. A main excluded fraction (fraction 1) was detected in all the samples examined. A part of this fraction was also excluded from Sepharose 4B. The molecular weight of this fractions can be assumed to be between 2·10 5 and 5·10 6. In some samples of saliva, one or two group-specific subfractions were identified (fractions 2 and 3). Fraction 2 is excluded from Sephadex G-50 and probably has a molecular weight between 10 000 and 13 000. Fraction 3 is dialysable and has a lower molecular weight, probably no greater than 1500–2000. Fraction 1 is water-soluble and can be precipitated in alcohol, while fractions 2 and 3 are both water- and alcohol-soluble.

Method for Typing Antisera to Herpesvirus hominis by Indirect Hemagglutination Inhibition

A test for typing antisera to Herpesvirus hominis that uses the method of indirect hemagglutination inhibition is described. The test, which is based upon the differential absorption of herpes antisera by preparations of type 1 and type 2 antigens, is rapidly and easily performed. The results permit some conclusions to be drawn regarding the antigenic relationships between the two virus types. Some of the practical limitations of the test are discussed.

ABO and Lewis Blood Grouping with ABH Secretor and Non-secretor Status: A Cross Sectional Study in Dhaka

A cross sectional study was done with 42 apparently healthy persons aged 6 years and above from both sexes. Most of them are blood donors in the department of Transfusion Medicine, Bangabandhu Sheikh Mujib Medical University (BSMMU), Dhaka, Bangladesh. Few, other than blood donor, were selected from the same locality. Five ml venous blood was collected with all aseptic precautions. ABO blood grouping and Lewis phenotyping were done by tube method. ABO reverse grouping was also done from serum. With all precautions 2 ml of saliva was collected from all subjects. Secretor status was detected from the saliva by haemagglutination inhibition method. ABO blood grouping shows 36% 'O' group, 24% 'A' group, 33% 'B' group and 7% 'AB' group. Distribution of Lewis phenotype are Le(a+b-) 19%, Le(a-b+) 53%, Le(a-b-) 26% and Le(a+b+) 2% only. 60% of study population was ABH secretor and 40% non-secretor. Keywords: ABO group; Lewis phenotype; ABH secretor status DOI: 10.3329/fmcj.v6i1.7409 Faridpur Med. Coll. J. 2011;6(1): 38-40

Detection of Fibrinogen Degradation Products by Use of Antibody Coated Latex Particles

SummaryA method for rapid determination of fibrinogen split products in human blood basing on agglutination of antibody coated latex particles is described. It’s sensitivity corresponds to that of Ouchterlony plates (17 μg/ml plasma). The result is available within 1 hr after drawing of blood. The method is semiquantitative with a at least as good reproducibility as the haemagglutination inhibition method. The reaction kinetics follow the principle of optimal proportion in a precipitin system.46 healthy people and 36 patients with various diseases were tested with the described method. 3 controls and 12 patients gave positive agglutination reactions that could not be attributed to the presence of split products. The mechanism of these side reactions which could be distinguished from the reactions due to fibrinogen degradation products is discussed. In the plasma of 10 patients split products could be demonstrated by the latex agglutination technic which was also successfully used to control the course of chronic intravascular coagulation in a patient with disseminated cancer of stomach. On account of the obtained results the method can be recommended as a fairly sensitive screening test for rapid bedside detection of fibrinogen degradation products.

Frequency of ABH secretors and non secretors: A cross sectional study in Karachi.

Objective: ABO blood group and secretor status is valuable in relation to some diseases in clinical and forensic medicine. Across the globe there are geographic and racial differences in the frequency of secretors and non-secretors. Aim of this study was to evaluate the status of ABH blood group secretors and non-secretors in Karachi (Pakistan). Methods: Blood and saliva samples were randomly collected from one hundred and one (n=101) healthy adult students (76 male, 25 female) ranging in age from 15 to 40 years. Their ABO and Rhesus blood groups were determined by conventional methods, and their secretor status was studied by hemagglutination inhibition method of saliva. Results: Results showed that 64.4% of the study population were ABH blood group secretors while 35.6% were non-secretors. Frequencies of the secretor status among various ABO blood groups were 71.4% in group A, 79.5% in group B, 45.5% in group AB, and 61.5% in group O. Conclusion: Frequency of ABH secretor is high (64.4%). Blood group B has the highest secretor (79.5%) frequency while Blood group AB has the lowest (45.5%).

Chorionic gonadotropin tests for diagnosis of pregnancy and chorionic tumors.

Various chorionic gonadotropin tests (HCG) for the diagnosis of pregnancy and chorionic tumors are reviewed. The tests divide into biological and immunological groups. Immunological methods are generally more sensitive than biological methods. A brief description of the widely used hemagglutination inhibition and latex particle agglutination methods is given. The antigen-antibody reaction which is involved in these methods has been applied in the development of highly sensitive radioimmunoassay procedures for detecting HCG. HCG levels during pregnancy rise very rapidly and reach a peak between Weeks 8-10 and decline sharply between Weeks 12-15 to a fairly constant level. The mean peak value of HCG in pregnancy has been established at 100000 IU 60-80 days after last menopause with a subsequent decline to about 20000 IU. Values of 300000 IU of HCG have been reported to occur in women with hydatid mole and instances as high as 3 million IU have been reported. The presence of titers of more than 200000 IU should indicate possible hydatid mole or choriocarcinoma though such values can occur in multiple pregnancy.

Measurement of Rubella Antibody in Immunoglobulin: its Disappearance from the Blood after Injection: Report of the Public Health Laboratory Service Working Party on Rubella

Rubella antibody titrations were done on samples of human immunoglobulin by neutralization and haemagglutination-inhibition methods. No significant variation was found in the antibody content of different batches. The specificity of the methods was confirmed by tests on a batch of human globulin specially prepared from plasma samples lacking rubella antibody.Divided doses of immunoglobulin were given to volunteers who had no rubella antibody. Low titres were then detected in the blood for a limited period and the disappearance of this antibody was followed.

Immunological studies of the luteinizing hormone: II. Comparative study of antigenic compounds

Comparison of the antigenic compounds in luteinizing hormone preparations of ovine, bovine, equine, and human origin was made using a hyperimmune rabbit antiserum to ovine luteinizing hormone. Methods involving agar double diffusion and immunoelectrophoresis revealed more precipitin lines in the ovine and bovine preparations than in those of either human or equine origin. The lines of the ovine and bovine hormones were fairly well balanced in electrophoretic mobility and similar to each other in appearance, whereas the lines of the equine and human hormones were predominantly anodic and differed in appearance from those of ovine and bovine preparations. The hemagglutination inhibition method showed that larger quantities of human and equine hormones than of ovine and bovine hormones were needed to inhibit the interaction between the hyperimmune antiserum and its homologous hormone. There was no correlation between the results of these tests and biological potency. Contamination with follicle-stimulating hormone was demonstrated by immunoprecipitation tests in all five hormone preparations, suggesting a lack of species specificity for this hormone and pointing to the clear antigenic dissimilarity between luteinizing and follicle stimulating hormones. Contamination with thyroid-stimulating hormone was demonstrated serologically only in the extracts of ovine and bovine glands. The ovine, bovine, and equine preparations of luteinizing hormone contained antigenic components which overlapped 3 3 An overlapping component is defined as a component certain antigenic determinants of which are shared by another hormone. with pregnant mare's serum gonadotropin, and the existence of a component in ovine, bovine, and human extracts which overlapped with human chorionic gonadotropin was also established.

Accuracy of a passive hemagglutination inhibition method for the determination of chorionic gonadotropin in the urine

This test (Prepuerin) when performed as described as a qualitative test for chorionic gonadotropin was uniformly negative or 100 per cent accurate on urines from 333 patients with 12 weeks or less of amenorrhea who were not pregnant. With urine from 537 similar patients who were pregnant the accuracy increased from 50 per ceni correct on urine front patients 5 weeks (29-35 days) to 85 then 95, 98, 99, 100, 100, and 100 per cent in the sixth through the twelfth week from the first day of their last menstrual period. On urine from patients with gynecologic complaints and who might be pregnant, the test was 97 per cent accurate over-all being truly positive in 120 of 121 positive tests and truly negative in 159 of 166 negative tests.

Quantitative immunologic assay of human chorionic gonadotropin in normal and abnormal pregnancies

Assays of urinary human chorionic gonadotropin (HCG) in normal and abnormal pregnancies performed by a hemagglutination inhibition method are presented. Antisera were produced in rabbits. An HCG preparation of pooled urine of pregnant women was used as a laboratory standard. 1st morning urine samples were obtained from a large group of patients mostly in aid of diagnosis of pregnancy. Patients with a variety of pathologic conditions also supplied specimens. There were a total of 1021 assays on 691 patients. It was possible to follow only 492 patients for 588 assays. There were 10 false positive and 2 false negative results for an overall accuracy of 98.0%. The mean level of HCG in normal pregnancies peaked at about 70,000 IU in the first trimester then fell to a constant level of 20,000 IU during the 2nd half of the pregnancy. The 95% confidence limits of the levels in normal pregnancy were from 10,000 to 350,000 IUD in the latter part of the first trimester and 2500-150,000 in the remainder. The levels of HCG in patients with clinical evidence of threatened abortion were determined. There were 86 such pregnancies, 47 of which had spontaneous abortion, while the remaining 39 went to term. The mean levels of the urinary HCG in the patients that aborted were consistently lower than those with normal pregnancies. Only during the period of 57-110 days after the last menstrual period were the values significant (P less than .001). 4 out of 6 patients with hydatidiform moles had titers of HCG greater than the normal range. HCG tests are also useful in following patients treated with chemotherapy for choriocarcinoma. The immunologic assay of HCG is a rapid, uncomplicated method of determining the level of HCG. It is of value in aiding the clinician in diagnosis, prognosis, and management of certain pathologic conditions of pregnancy.

Daily urinary assay of luteinizing hormone by an immunologic method

1.1. Immunologic cross-reaction between HCG and LH has been demonstrated using an hemagglutination inhibition method.2.2. Daily urinary assay of LH by an immunologic method in a group of normal menstruating women shows a midcycle rise of LH about the time of ovulation. These findings agree with those previously published using biologic assay methods.3.3. LH assays on a small group with ovulatory failure show different patterns of excretion. The possibilities of further studies with this method are discussed.