scholarly journals Application of the enzyme linked immunospecific assay (ELISA) for the detection of platelet antibodies

Blood ◽  
1981 ◽  
Vol 57 (1) ◽  
pp. 32-37
Author(s):  
M Gudino ◽  
WV Miller
Keyword(s):  
Color Change ◽  
Serum Antibody ◽  
Enzyme Reaction ◽  
Antibody Concentration ◽  
Immunofluorescence Test ◽  
Platelet Antibodies ◽  
Positive Results ◽  
Assay Conditions ◽  
Enzyme Alkaline Phosphatase ◽  
Platelet Antibody

Many methods have been described to identify platelet antibody, but they are either not very sensitive or too complex for general use. Therefore, we have developed an enzyme immunoassay for the detection of platelet antibodies in serum. The method involves incubating platelets with serum antibody; any attached antibody is shown by the addition of an enzyme (alkaline phosphatase) labeled anti-human IgG, followed by assay of the enzyme reaction with its substrate. The reaction product is indicated by a color change, which is proportional to the antibody concentration. Assay conditions such as the use of paraformaldehyde fixed versus unfixed platelets, conjugate dilutions, and substrate concentration and incubation time were investigated. Positive results were obtained in 16 of 19 sera of patients with various diseases including 2 of 4 patients with idiopathic thrombocytopenic purpura, 2 of 2 with post-transfusion purpura, 2 of 3 with neonatal purpura, and all 9 polytransfused patients. Sensitivity and specificity were 84% and 98%, respectively. Also, enzyme linked immunospecific assay (ELISA) was found to be superior to the lymphocytotoxicity (LCT) and platelet immunofluorescence test (PIIFT) for platelet antibody identification.

scholarly journals Application of the enzyme linked immunospecific assay (ELISA) for the detection of platelet antibodies

Blood ◽  
1981 ◽  
Vol 57 (1) ◽  
pp. 32-37 ◽  
Author(s):  
M Gudino ◽  
WV Miller
Keyword(s):  
Color Change ◽  
Serum Antibody ◽  
Enzyme Reaction ◽  
Antibody Concentration ◽  
Immunofluorescence Test ◽  
Platelet Antibodies ◽  
Positive Results ◽  
Assay Conditions ◽  
Enzyme Alkaline Phosphatase ◽  
Platelet Antibody

Abstract Many methods have been described to identify platelet antibody, but they are either not very sensitive or too complex for general use. Therefore, we have developed an enzyme immunoassay for the detection of platelet antibodies in serum. The method involves incubating platelets with serum antibody; any attached antibody is shown by the addition of an enzyme (alkaline phosphatase) labeled anti-human IgG, followed by assay of the enzyme reaction with its substrate. The reaction product is indicated by a color change, which is proportional to the antibody concentration. Assay conditions such as the use of paraformaldehyde fixed versus unfixed platelets, conjugate dilutions, and substrate concentration and incubation time were investigated. Positive results were obtained in 16 of 19 sera of patients with various diseases including 2 of 4 patients with idiopathic thrombocytopenic purpura, 2 of 2 with post-transfusion purpura, 2 of 3 with neonatal purpura, and all 9 polytransfused patients. Sensitivity and specificity were 84% and 98%, respectively. Also, enzyme linked immunospecific assay (ELISA) was found to be superior to the lymphocytotoxicity (LCT) and platelet immunofluorescence test (PIIFT) for platelet antibody identification.

scholarly journals Serum Concentration of Antibodies to Mumps, but Not Measles, Rubella, or Varicella, Is Associated with Intake of Dietary Fiber in the NHANES, 1999–2004

Nutrients ◽  
2021 ◽  
Vol 13 (3) ◽  
pp. 813
Author(s):  
Cynthia B. Van Van Landingham ◽  
Debra R. Keast ◽  
Matthew P. Longnecker
Keyword(s):  
Measurement Error ◽  
Dietary Fiber ◽  
Meta Analysis ◽  
Serum Antibody ◽  
Potential Effect ◽  
Interquartile Range ◽  
Antibody Concentration ◽  
Fiber Intake ◽  
Food Groups ◽  
Percentage Difference

Treatment with prebiotics, a type of dietary fiber, was recently shown to increase antibody concentrations following influenza vaccination in a meta-analysis of clinical trials. In observational epidemiologic studies it is not possible to estimate intake of prebiotics, but quantifying intake of dietary fiber is routine. Our objective was to investigate the potential effect of dietary fiber on immunogenicity. We examined serum antibody concentrations (Measles, Mumps, Rubella, and Varicella) in relation to dietary fiber in more than 12,000 subjects in the U.S. National Health and Nutrition Examination Survey (NHANES) for the period 1999–2004. Data from one (1999–2002) or two (2003–2004) dietary recalls were used to calculate fiber intake. For Mumps the adjusted percentage difference in antibody concentration per interquartile range intake in energy-adjusted dietary fiber was 6.34% (95% confidence interval, 3.10, 9.68). Fiber from grain-based foods was more positively associated than fiber from other fiber-containing food groups. The association was slightly larger among subgroups with higher fiber intake, greater interquartile range in fiber intake, and less measurement error. Furthermore, based on the reliability of the diet recalls in 2003–2004, we calculated that the percentage difference per interquartile increment was substantially attenuated by measurement error. Dietary fiber may have a favorable influence on the immunogenicity of some vaccines or natural infections.

Responses of Children Immunized with Capsular Polysaccharide of Hemophilus influenzae Type b, by David H. Smith, MD, et al,Pediatrics, 1973;52:637–644; and Haemophilus influenzae Type b Capsular Polysaccharide Vaccine in Children: A Double-blind Field Study of 100 000 Vaccinees 3 Months to 5 Years of Age in Finland, by Heikki Peltola, MD et al,Pediatrics, 1977;60:730–737

PEDIATRICS ◽  
1998 ◽  
Vol 102 (Supplement_1) ◽  
pp. 252-254
Author(s):  
Georges Peter
Keyword(s):  
Capsular Polysaccharide ◽  
Serum Antibody ◽  
Polysaccharide Vaccine ◽  
Haemophilus Influenzae Type ◽  
Antibody Concentration ◽  
Antigen Binding ◽  
Haemophilus Influenzae Type B ◽  
Type B ◽  
Age Dependent ◽  
Group A

One hundred forty-one children of 5 to 59 months of age were immunized with a single intramuscular dose of 0.67, 3.3, 17, or 67 μg polyribophosphate (PRP), the capsular antigen ofHemophilus influenzae, type b. The immunizations were well tolerated, particularly at doses of .67 to 17 μg. Antibody activity was measured by radioactive antigen binding, using3H-labelled PRP. Doses of 3.3 and 17 μg produced significant antibody rises in nearly 90% of recipients; 0.67 and 67 μg in approximately half. The geometric mean titers were similar at three and six weeks after immunization and were greater with the middle doses. The net antibody increase in responding children was strongly age dependent, but was not related to the preimmunization antibody concentration. Rises in serum bactericidal activity against H. influenzae type b generally accompanied rises in antibody concentration as measured by the antigen-binding assay. A recently developed Haemophilus influenzae type b capsular polysaccharide vaccine was given to 48 977 children 3 months to 5 years of age; an equal number of children receiving group A meningococcal vaccine served as controls. The protection as well as serum antibody response was strongly age dependent. Among children who had received the H. influenzae type b vaccine when 18 months of age or older, there were no cases of bacteremic disease caused by H. influenzaetype b in the first year after vaccination. At the same time 11 such cases were seen in the control group of the same age, a highly significant difference. In the second year after vaccination two cases occurred in the H. influenzae type b-vaccinated group, five in the meningococcal-group A vaccinated group. No protection was seen among children who had been younger than 18 months when vaccinated, even if they received a booster dose of the vaccine. The serum antibody response to the H. influenzae type b polysaccharide, measured by radioimmunoassay, was poor in children below 18 months of age and good in those above it. No effect of the vaccine could be seen on the nasopharyngeal carriage of H. influenzae type b, which was approximately 6% in this age group. Adverse effects of the vaccine were mild.

Immunogenicity of Haemophilus influenzae Type b Polysaccharide-Neisseria meningitidis Outer Membrane Protein Conjugate Vaccine in 2- to 6-Month-Old Infants

PEDIATRICS ◽  
1987 ◽  
Vol 80 (2) ◽  
pp. 283-287
Author(s):  
Allen A. Lenoir ◽  
Paul D. Granoff ◽  
Dan M. Granoff
Keyword(s):  
Membrane Protein ◽  
Haemophilus Influenzae ◽  
Neisseria Meningitidis ◽  
Outer Membrane Protein ◽  
Geometric Mean ◽  
Serum Antibody ◽  
Haemophilus Influenzae Type ◽  
Antibody Concentration ◽  
Haemophilus Influenzae Type B ◽  
Type B

Fifty infants, 2 to 6 months of age, were vaccinated with Haemophilus influenzae type b capsular polysaccharide covalently linked to an outer membrane protein from Neisseria meningitidis group B. Subjects were given two injections and were randomly assigned to receive the injections separated by 1 or 2 months. Each dose contained 15 µg of polysaccharide and 51 µg of protein, or approximately twice the amount of polysaccharide as used in our previous trial (Lancet 1986;2:299). Fevers of 38.0° to 38.8°C developed in three infants (6%) within 24 hours after vaccination, but there were no other notable reactions. Following one injection, the geometric mean antibody concentration increased from 0.13 µg/mL in preimmune serum to 1.50 µg/mL in serum obtained 1 to 2 months later (P < .001). After a second injection, there was a further increase in serum antibody (geometric mean = 3.11 µg/mL, P < .007). The geometric mean antibody concentration of the group reimmunized 2 months after the first injection was higher than that in the group reimmunized after 1 month (3.95 v 2.32 µg/mL, P = .05, by analysis of covariance with age as the covariant). These data confirm our previous preliminary observations on the safety and immunogenicity of this new conjugate vaccine in infants 2 to 6 months of age. The data suggest that a 2-month interval between the first and second injections results in higher levels of serum antibody than a 1-month interval.

Interchangeability of Haemophilus influenzae Type b vaccines in the Primary Series: Evaluation of a Two-dose Mixed Regimen

PEDIATRICS ◽  
1996 ◽  
Vol 98 (5) ◽  
pp. 898-904 ◽  
Author(s):  
Kathleen M. Bewley ◽  
Joel G. Schwab ◽  
Gerard A. Ballanco ◽  
Robert S. Daum
Keyword(s):  
Haemophilus Influenzae ◽  
Randomized Clinical Trials ◽  
Geometric Mean ◽  
Serum Antibody ◽  
Area Under The Curve ◽  
Haemophilus Influenzae Type ◽  
Antibody Concentration ◽  
Haemophilus Influenzae Type B ◽  
Type B ◽  
Series Evaluation

Objective. To evaluate two- or threedose "mixed" regimens of Haemophilus influenzae type b conjugate vaccines in the priming series. Design. Two randomized clinical trials with 140 and 181 infants, respectively. Setting. Private practices in New Orleans and Chicago. Methods. In trial I, infants received one of four regimens. Two were recommended regimens for polyribosylribitol phosphate (PRP)—meningococcal protein conjugate (M) and PRP—tetanus toxoid conjugate (T). Two mixed regimens consisted of M at 2 months followed by two doses of T or PRP—diphtheria toxoid conjugate (D) at 4 and 6 months. Trial II consisted of three groups. Two were recommended regimens for M and T. The third was a two-dose mixed regimen consisting of M at 2 months and T at 4 months. Parents were interviewed and instructed to record side effects after each vaccination. Serum was assayed for H influenzae type b anticapsular antibody (anti-PRP). Results. Minor differences in safety profiles likely reflected α error. In trial I, M (lot 0884T, one of several known to have had decreased immunogenicity), probably primed for substantial increase in serum antibody when D or T was given at 4 and 6 months. In trial II, infants who received the two-dose mixed regimen (M from immunogenic lot 0116W at 2 months and T at 4 months) had a significantly higher mean area under the curve than recipients of the three-dose TIT regimen when antibody concentration was plotted against age, although the geometric mean anti-PRP antibody concentration for the MT-recipients was significantly lower at 7 months. Conclusions. M used in trial I may have primed infants despite poor immunogenicity. The two-dose mixed regimen (MT-) in trial II produced a mean anti-PRP antibody concentration with higher sustained anti-PRP concentrations from 2 to 7 months, as judged by the area under the curve, but a lower mean anti-PRP antibody concentration at 7 months.

scholarly journals Behaviour of indirect immunofluorescence test and some cerebrospinal parameters in neurocysticercosis

1987 ◽  
Vol 45 (1) ◽  
pp. 33-43 ◽  
Author(s):  
Amauri Braga Simonetti ◽  
Jorge Teixeira
Keyword(s):  
Cerebrospinal Fluid ◽  
Indirect Immunofluorescence ◽  
False Positive ◽  
Clinical Evidence ◽  
Routine Analysis ◽  
Immunofluorescence Test ◽  
Indirect Immunofluorescence Test ◽  
Immunofluorescent Test ◽  
Positive Results

Cerebrospinal fluid from 53 patients with clinical evidence of neurocysticercosis and 11 who suffered from several diseases were studied to evaluate the behaviour of indirect immunofluorescence test and some parameters of routine analysis. In neurocysticercosis there were pleocitosis in 88.7% of cases, eosinophilorrachia in 60.3%o, hyperproteinorrachia in 71.7% and hypoglucorrachia in 13.2%. The indirect immunofluorescence test was positive in 19.2% of cases but false-positive results were found when the samples showed xanthochromia or erythrocyte contamination. The authors discuss their results in comparison with those in literature and conclude that the immunofluorescent test is sensitive and useful in diagnosis of neurocysticercosis, except when the interferents previously mentioned are present.

scholarly journals Pseudothrombocytopenia: an immunologic study on platelet antibodies dependent on ethylene diamine tetra-acetate

Blood ◽  
1982 ◽  
Vol 59 (1) ◽  
pp. 157-161 ◽  
Author(s):  
JG Pegels ◽  
EC Bruynes ◽  
CP Engelfriet ◽  
AE von dem Borne
Keyword(s):  
Ethylene Diamine ◽  
Igg Antibodies ◽  
Immunofluorescence Test ◽  
Igm Antibodies ◽  
Platelet Antibodies ◽  
Group Specificity ◽  
Iga Antibodies ◽  
Ethylene Diamine Tetra Acetate

Abstract Antibodies specifically reacting with platelets only in the presence of EDTA, by the platelet immunofluorescence test, were found in the serum of 20 patients with pseudothrombocytopenia due to in vitro EDTA- dependent platelet agglutination. These antibodies reacted optimally at 0–4 degree C. In 19 patients, IgG antibodies were detected; in 8 patients, IgM or IgA antibodies were also found. In one patient, only IgM antibodies were found. In 14 patients, the IgG antibodies were IgG1, but IgG2, IgG3, and IgG4 antibodies were also seen in 7 patients. The reaction of platelets with the antibodies was detectable in the presence of Na2EDTA, the K, Ca, and Mg salts of EDTA, and K2EGTA. F(ab')2 or F(ab') fragments of the IgG antibodies reached as strongly as the intact antibodies, indicating that the reaction is dependent on the antibody-combining site. The EDTA-dependent antibodies did not show platelet-group specificity. However, platelets from patients with Glanzmann disease did not react with the antibodies.

scholarly journals AET-treated platelets: their usefulness for platelet antibody detection and an examination of their altered sensitivity to immune lysis

Blood ◽  
1979 ◽  
Vol 54 (5) ◽  
pp. 1101-1108
Author(s):  
PL Cimo ◽  
SA Gerber
Keyword(s):  
Blood Cells ◽  
Paroxysmal Nocturnal Hemoglobinuria ◽  
Antibody Detection ◽  
Enhanced Sensitivity ◽  
Platelet Antibodies ◽  
Different Types ◽  
Drug Dependent ◽  
Altered Sensitivity ◽  
Bernard Soulier Syndrome ◽  
Platelet Antibody

2-Aminoethylisothiouronium bromide (AET) increases the sensitivity of blood cells to complement-mediated immune lysis. We compared the sensitivities of untreated or AET-treated platelets to immune lysis induced by different types of platelet antibody in the 51Cr platelet lysis test. AET platelets were 8–16 times more sensitive to autoantibody and alloantibody, but 8–16 times less sensitive to drug- dependent antibody. AET-platelets bound similar amounts of alloantibody but less drug-dependent antibody, and they lysed at higher complement dilutions than did untreated platelets. AET-platelets detected 10 of 25 autoantibodies, 9 of 9 alloantibodies, and 5 of 8 drug-dependent antibodies. Untreated platelets detected 1 of 25, 6 of 9, and 7 of 8 of these respective platelet antibodies. The use of AET-platelets in the 51Cr platelet lysis test increases its sensitivity for detecting non- drug-dependent platelet antibodies. AET-platelets resemble paroxysmal nocturnal hemoglobinuria (PNH) platelets in their enhanced sensitivity to complement-mediated lysis. They differ from PNH platelets in their insensitivity to immune lysis induced by drug-dependent antibodies and, in this respect, are similar to Bernard-Soulier syndrome platelets.

Effects of Splenectomy on Immune Thrombocytopenic Purpura in (NZW x BXSB) F1 Mice: Analyses of Platelet Kinetics and Anti-Platelet Antibody Production

1992 ◽  
Vol 67 (05) ◽  
pp. 563-566 ◽  
Author(s):  
Hajime Mizutani ◽  
Takayasu Furubayashi ◽  
Hirokazu Kashiwagi ◽  
Shigenori Honda ◽  
Hironori Take ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Antibody Production ◽  
Immune Thrombocytopenic Purpura ◽  
Therapeutic Agents ◽  
Thrombocytopenic Purpura ◽  
Platelet Counts ◽  
Platelet Antibodies ◽  
Splenic Uptake ◽  
Normal Controls ◽  
Platelet Antibody

SummaryEffects of splenectomy on platelet kinetics and production of anti-platelet antibodies were studied in male (NZW × BXSB) F1 (W/B F1) mice, which are known as the animal model of immune thrombocytopenic purpura (ITP). Studies on organ localization of radiolabeled platelets revealed that splenic uptake significantly increases in W/B F1 mice in comparison with that of normal controls. W/B Fj mice showed a significant increase in platelet counts and, in contrast with sham-operated controls, high levels of platelet counts were maintained up to 6 weeks after splenectomy. Platelet lifespans (PLSs) did not reach normal levels, although prolonged PLSs were observed. In addition, platelet-associated antibody (PAA) values showed a tendency towards transient decrease, but there was no change in platelet-bindable serum antibodies (PBAs). These findings indicate that the suppression of anti-platelet antibody production is essential to the treatment of ITP; splenectomy may not be effective in treating severely affected ITP patients because, although the spleen is one of the major sites of platelet sequestration and antibody production, reticulo-endothelial systems (RESs) (liver, bone marrow, lymphnodes, etc.) other than the spleen are also responsible for the destruction of platelets. We therefore consider the W/B F1 mouse to be a useful model of human ITP, and believe that it provides valuable information for the development of new therapeutic agents in patients with ITP, especially those who do not respond to splenectomy.

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