scholarly journals Detection of platelet antibodies using a micro-enzyme-linked immunosorbent assay (ELISA)

Blood ◽  
1983 ◽  
Vol 61 (2) ◽  
pp. 311-317 ◽  
Author(s):  
CA Schiffer ◽  
V Young
Keyword(s):  
Immunosorbent Assay ◽  
Immune Thrombocytopenic Purpura ◽  
Hla Typing ◽  
Enzyme Linked Immunosorbent Assay ◽  
Thrombocytopenic Purpura ◽  
Microtiter Plates ◽  
Platelet Antibodies ◽  
Liquid Suspension ◽  
Large Numbers ◽  
Specific Antigens

Abstract An enzyme-linked immunosorbent assay (ELISA) for the measurement of circulating platelet antibody and platelet-associated IgG (PAIgG) is described. The test is done in microtiter plates and rapidly provides quantitative and highly reproducible results. Alloantibodies from 28 of 30 multiple transfused patients and isoantibodies from 3 of 4 patients with immune thrombocytopenic purpura (ITP) were detected. PAIgG was elevated in all 4 patients with ITP, and HLA and platelet-specific antigens were reliably detected using HLA typing sera and anti-PIA1 antibody, respectively. Platelets preserved wither by dessication in the wells of the microtiter plates or in liquid suspension in saline at 4 degrees C gave results comparable to values using fresh platelets. Storage periods ranged from 30 days for dessicated platelets to more than 1 yr for platelets stored in suspension. The ability to utilize preserved platelets may allow relatively convenient screening of large numbers of potential platelet donors for alloimmunized patients.

Detection of platelet antibodies using a micro-enzyme-linked immunosorbent assay (ELISA)

Blood ◽  
1983 ◽  
Vol 61 (2) ◽  
pp. 311-317
Author(s):  
CA Schiffer ◽  
V Young
Keyword(s):  
Immunosorbent Assay ◽  
Immune Thrombocytopenic Purpura ◽  
Hla Typing ◽  
Enzyme Linked Immunosorbent Assay ◽  
Thrombocytopenic Purpura ◽  
Microtiter Plates ◽  
Platelet Antibodies ◽  
Liquid Suspension ◽  
Large Numbers ◽  
Specific Antigens

An enzyme-linked immunosorbent assay (ELISA) for the measurement of circulating platelet antibody and platelet-associated IgG (PAIgG) is described. The test is done in microtiter plates and rapidly provides quantitative and highly reproducible results. Alloantibodies from 28 of 30 multiple transfused patients and isoantibodies from 3 of 4 patients with immune thrombocytopenic purpura (ITP) were detected. PAIgG was elevated in all 4 patients with ITP, and HLA and platelet-specific antigens were reliably detected using HLA typing sera and anti-PIA1 antibody, respectively. Platelets preserved wither by dessication in the wells of the microtiter plates or in liquid suspension in saline at 4 degrees C gave results comparable to values using fresh platelets. Storage periods ranged from 30 days for dessicated platelets to more than 1 yr for platelets stored in suspension. The ability to utilize preserved platelets may allow relatively convenient screening of large numbers of potential platelet donors for alloimmunized patients.

Serum Thrombopoietin (TPO) Levels in Patients with Amegakaryocytic Thrombocytopenia Are much Higher than those with Immune Thrombocytopenic Purpura

1996 ◽  
Vol 76 (05) ◽  
pp. 675-678 ◽  
Author(s):  
Harumi Y Mukai ◽  
Hiroshi Kojima ◽  
Kazuo Todokoro ◽  
Tomoyuki Tahara ◽  
Takashi Kato ◽  
...  
Keyword(s):  
Correlation Coefficient ◽  
Immunosorbent Assay ◽  
Immune Thrombocytopenic Purpura ◽  
Steroid Treatment ◽  
Enzyme Linked Immunosorbent Assay ◽  
Thrombocytopenic Purpura ◽  
Platelet Counts ◽  
Healthy Donors

SummaryWe assayed serum thrombopoietin (TPO) levels in amegakaryocytic thrombocytopenia (AMT) and immune thrombocytopenic purpura (ITP) patients by using a newly established enzyme-linked immunosorbent assay (ELISA). TPO levels in AMT patients were quite high (mean ± SD = 13.7 ± 11.2 fmoles/ml, n = 4), whereas those in ITP patients were only slightly higher (1.25 ± 0.39, n = 12) than those of the healthy donors (0.55 ± 0.2, n = 20). Furthermore, in ITP patients no correlation was observed between platelet counts and serum TPO levels (correlation coefficient = 0.14). We further assayed serum TPO levels sequentially during steroid treatment in patients with AMT and ITP. In one AMT patient serum TPO levels started to decrease in accordance with the increase of megakaryocyte counts, which preceded the increase in platelet counts. However, in ITP patients serum TPO levels did not change significantly throughout the course of the treatment despite the recovery of platelet counts. Based on these findings, we conclude that serum TPO levels may be regulated at least in part by megakaryocyte counts.

An enzyme-linked immunosorbent assay for the detection of Rathayibacter toxicus, the bacterium involved in annual ryegrass toxicity, in hay

2006 ◽  
Vol 57 (7) ◽  
pp. 731 ◽  
Author(s):  
A. M. Masters ◽  
A. R. Gregory ◽  
R. J. Evans ◽  
J. E. Speijers ◽  
S. S. Sutherland
Keyword(s):  
Monoclonal Antibody ◽  
Immunosorbent Assay ◽  
Specific Antigen ◽  
Cost Effective ◽  
Enzyme Linked Immunosorbent Assay ◽  
Annual Ryegrass ◽  
Capture Assay ◽  
Large Numbers ◽  
Soluble Polysaccharide ◽  
Annual Ryegrass Toxicity

An enzyme-linked immunosorbent assay (ELISA) for Rathayibacter toxicus is described. The development of a monoclonal antibody for a specific antigen from R. toxicus and a polyclonal antibody raised against the same R. toxicus preparation enabled a capture assay format. The assay is specific for a soluble polysaccharide produced by the bacterium and was found to be sensitive enough to detect antigen equivalent to less than one gall per kilogram of hay. The applicability of the assay to samples of pasture or hay is demonstrated. Cost-effective testing of large numbers of samples for the presence of R. toxicus is possible with the ELISA. This will assist stockowners, hay producers, and hay exporters in the management of the risk of annual ryegrass toxicity.

scholarly journals Reevaluation of enzyme-linked immunosorbent assay with ultraviolet-treated polystyrene microtiter plates for measurement of antibodies to dsDNA.

1996 ◽  
Vol 19 (2) ◽  
pp. 163-167
Author(s):  
Junichi Kaburaki ◽  
Takashi Ogasawara ◽  
Masakatsu Hayakawa ◽  
Masataka Kuwana ◽  
Takeshi Tojo ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Enzyme Linked Immunosorbent Assay ◽  
Microtiter Plates ◽  
Antibodies To Dsdna

Regulation of Serum Thrombopoietin Levels by Platelets and Megakaryocytes in Patients with Aplastic Anaemia and Idiopathic Thrombocytopenic Purpura

1996 ◽  
Vol 76 (02) ◽  
pp. 156-160 ◽  
Author(s):  
Naoaki Ichikawa ◽  
Fumihiro Ishida ◽  
Shigetaka Shimodaira ◽  
Tomoyuki Tahara ◽  
Takashi Kato ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Platelet Count ◽  
Aplastic Anaemia ◽  
Idiopathic Thrombocytopenic Purpura ◽  
Immunosorbent Assay ◽  
Regulatory Mechanism ◽  
Inverse Correlation ◽  
Normal Subjects ◽  
Enzyme Linked Immunosorbent Assay ◽  
Thrombocytopenic Purpura

SummaryTo clarify the regulatory mechanism of thrombopoietin (TPO, c-Mpl ligand) in chronic thrombocytopenic conditions, we determined TPO levels in the sera of patients with aplastic anaemia (AA; n = 26) and idiopathic thrombocytopenic purpura (ITP; n = 32) by an enzyme-linked immunosorbent assay. Despite a similarity in platelet counts, serum TPO levels in the AA group were markedly higher than those in the ITP group: 20.41 ± 9.71 f mol/ml (mean ± SD) and 1.66 ± 0.55 f mol/ml, respectively, both of which were significantly elevated compared to normal subjects (n = 41; 1.22 ± 0.37). In both groups, serum TPO level showed an inverse correlation with the platelet count. We determined the megakaryocyte volume using bone marrow clot section and found that it was markedly small in the AA group; while in the ITP group it was augmented with a correlation to serum TPO level. Our findings suggest that TPO levels may be regulated not only by platelets but also megakaryocytes in AA and ITP.

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.

scholarly journals An enzyme-linked immunosorbent assay for platelet compatibility testing

Blood ◽  
1983 ◽  
Vol 62 (4) ◽  
pp. 744-749 ◽  
Author(s):  
JD Tamerius ◽  
JG Curd ◽  
P Tani ◽  
R McMillan
Keyword(s):  
Antibody Titer ◽  
Immunosorbent Assay ◽  
Clinical Laboratory ◽  
Clinical Problem ◽  
Enzyme Linked Immunosorbent Assay ◽  
Microtiter Plates ◽  
Volunteer Donor ◽  
Antibody Titers ◽  
Test Serum ◽  
Selection Of

Abstract The selection of platelet donors for patients who are refractory to random donor platelets often presents a difficult clinical problem. We describe an enzyme-linked immunosorbent assay (ELISA) for evaluating alloantibodies in refractory patients. Platelets from prospective donors are immobilized on microtiter plates and, after incubation with test serum and washing, platelet-bound IgG is detected with enzyme- linked anti-human IgG. Platelets from 46 prospective donors were tested. Twenty-two were judged compatible (reciprocal of the antibody titer less than 16) and, of these, 15 were used as platelet donors; each gave a measurable platelet increment after transfusion. The magnitude of the response was roughly proportional to the assay results. Platelets from donors giving antibody titers less than 4 resulted in platelet increments at 1 hr ranging from 4,890 to 22,200 (median 12,600), while platelets from donors giving titers of 8 or 16 resulted in lesser increments (550–4548). Conversely, 5 of the 24 patients found incompatible by the assay (titer greater than 16) gave no platelet increment, and in 3 instances, the recipient developed fever and chills after the transfusion. The assay is sensitive, simple, and adaptable to the clinical laboratory. Platelets from volunteer donor panels can be plated and stored for up to 6 mo.

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