Detection and identification of platelet antibodies using a sensitive multiplex assay system-platelet antibody bead array

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.

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

Thrombosis and Haemostasis ◽

10.1055/s-0038-1648494 ◽

1992 ◽

Vol 67 (05) ◽

pp. 563-566 ◽

Cited By ~ 12

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

Blood ◽

10.1182/blood.v57.1.32.32 ◽

1981 ◽

Vol 57 (1) ◽

pp. 32-37 ◽

Cited By ~ 31

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.

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

Blood ◽

10.1182/blood.v54.5.1101.1101 ◽

1979 ◽

Vol 54 (5) ◽

pp. 1101-1108 ◽

Cited By ~ 5

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

Abstract 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.

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Rapid detection and identification of viral and bacterial fish pathogens using a DNA array-based multiplex assay

Journal of Fish Diseases ◽

10.1111/j.1365-2761.2011.01304.x ◽

2011 ◽

Vol 34 (11) ◽

pp. 861-875 ◽

Cited By ~ 15

Author(s):

B Lievens ◽

I Frans ◽

C Heusdens ◽

A Justé ◽

S P Jonstrup ◽

...

Keyword(s):

Rapid Detection ◽

Multiplex Assay ◽

Dna Array ◽

Fish Pathogens ◽

Detection And Identification

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

Blood ◽

10.1182/blood.v57.1.32.bloodjournal57132 ◽

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.

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Evaluation of influence of IL-6 C-572G gene polymorphism and clinical factors on positive platelet antibody test

Journal of Advanced Pediatrics and Child Health ◽

10.29328/journal.japch.1001023 ◽

2021 ◽

Vol 4 (1) ◽

pp. 006-012

Author(s):

Lin Jeong-Shi ◽

Lee Li-Hsuan ◽

Liu Hsueng-Mei ◽

Chen Ying-Ju ◽

Chiou Tzeon-Jye

Keyword(s):

Gene Polymorphism ◽

Solid Phase ◽

Antibody Test ◽

Antibody Detection ◽

Nucleotide Polymorphisms ◽

Clinical Factors ◽

Hematologic Diseases ◽

Platelet Antibodies ◽

Positive Reactivity ◽

Platelet Antibody

Background: Interleukin-6 (IL-6) promotes antibody production. The objective of this study was to investigate whether IL-6 C-572G single nucleotide polymorphisms (SNP) and clinical factors are associated with positive platelet antibody test. Materials and methods: Thirty platelet recipients with platelet antibodies (responders) and 20 platelet recipients without platelet antibodies (non-responders) were randomly selected. The -572 C>G (rs 1800796) SNPs in the promoter region of IL-6 gene were genotyped by polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) method. Solid phase red cell adherence assay (SPRCA) was used for platelet antibody detection. Results: Age, sex, percentage patients with benign diseases, and percentage of patients with homozygotes for the C allele at position -572 of the IL-6 gene were similar between responders and non-responders. Although the amounts of platelets pheresis transfused to patients with hematologic diseases were higher than those of non-hematologic diseases (47.2 ± 54.2 vs. 17.4 ± 13.8 units, p = 0.019), detection rate of platelet antibodies was lower in patients with hematologic diseases than that in patients with non-hematologic diseases (42.3% vs. 79.2%, p = 0.01). Conclusion: There was no association between IL-6 C-572G gene polymorphism and positive reactivity in solid phase platelet antibody detection method in platelet recipients.

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