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CONTENTS
Immunohematology
Volume 13, Number 2, 1997

ABSTRACTS

Semiautomation of platelet HPA-1a phenotyping by SPRCA and ELISA
L.A. Mohabir and L. Porter

A clinically significant anti-HLA-A2 detectable by extended incubation cytotoxicity and flow cytometric techniques but not by a standard NIH lymphocytotoxicity test
S.F. Garner, J. Petrochilos, C.J. Brown, S. Cavanna, I. Chanarin, and C. Navarrete

Delayed hemolytic transfusion reaction and paroxysmal cold hemoglobinuria: an unusual association
M.A. Wodzinski, R.C. Collin, D.J. Booker, F. Stamps, J.D Bellamy, and R.J Sokol

Case report: reporting anti-G as anti-C+D may have misleading clinical implications
A.L. Mosley, M.B. Trich, N.C. Thomas, and S.G.Sandler

A case of hemolytic disease of the newborn due to anti-Kpa
L. Costamagna, M. Barbarini, G.L. Viarengo, A. Pagani, D. Isernia, and L. Salvaneschi

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Semiautomation of platelet HPA-1a phenotyping by SPRCA and ELISA

L.A. Mohabir and L. Porter

An enzyme-linked immunosorbent assay (ELISA) and solid phase red cell adherence assay (SPRCA) were assessed for platelet HPA-1a typing in U well microplates. Both methods were partially automated by the use of the Tecan RSP 8051ID robotic sampler and the SLT 400 ATC plate reader with Soft2000 software. Pretreatment of the adherent platelets with chloroquine diphosphate or citric acid enabled anti-HPA-1a, even when contaminated with HLA class 1 antibodies, to be used for typing. Of 675 antenatal samples, 13 were identified as HPA-1a negative. A further 36 known donor HPA-1a negative samples were correctly typed by both methods. Concordant results were obtained for the ELISA and SPRCA. All presumed HPA-1a negatives were confirmed by either polymerase chain reaction using sequence-specific primers (PCR-SSP) or by monoclonal antibody-specific immobilization of platelet antigens (MAIPA). The two semiautomated systems provide simple, accurate, reproducible, and secure ways for large scale HPA-1a phenotyping. The SPRCA can also be performed as a manual technique for small batches of samples.
Immunohematology1997;13:4448.

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A clinically significant anti-HLA-A2 detectable by extended incubation cytotoxicity and flow cytometric techniques but not by a standard NIH lymphocytotoxicity test

S.F. Garner, J. Petrochilos, C.J. Brown, S. Cavanna, I. Chanarin, and C. Navarrete

A previously transfused female patient, known to have a platelet defect, was transfused with platelets prior to surgery. After the 18th unit she felt unwell, developed fever, rigor, became nauseous, and vomited. Her blood pressure decreased from 140/90 to 80/50mm Hg. Passive transfer of donor granulocytes or red cell antibodies were excluded as a cause. Therefore, a serum sample from the patient was investigated for the presence of antibodies to human leukocyte antigens (HLA) using a standard National Institutes of Health (NIH) lymphocytotoxicity test, but antibodies were not detected. However, an extended incubation cytotoxicity test demonstrated the presence of an anti-HLA-A2, and indirect immunofluorescence flow cytometry showed the presence of an IgG1 antibody reacting with 50 percent of cells in a random pool of lymphocytes. One week later, multispecific HLA antibodies were detectable by both NIH and extended incubation cytotoxicity tests. Flow cytometry showed a 16-fold increase in the amount of IgG antibodies and the appearance of an IgM component. Such clinically important HLA antibodies can be detected by extended incubation cytotoxicity and flow cytometric assays prior to becoming reactive in a standard NIH cytotoxicity technique.
Immunohematology1997;13:4953.

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Delayed hemolytic transfusion reaction and paroxysmal cold hemoglobinuria: an unusual association

M.A. Wodzinski, R.C. Collin, D.J. Booker, R. Stamps, J.D. Bellamy, and R.J. Sokol

An 80-year-old female presented with melena and anemia due to bleeding from a benign gastric ulcer. Her blood group was O, D+. The serum contained anti-B and a weak anti-A (titer 2 at 18C). She was inadvertently transfused with approximately 3.5 units of group A red blood cells with no initial ill effects. One week later, the anti-A titer increased to 8 and the direct antiglobulin test (DAT) was weakly positive (IgG and C3d). The next day, intravascular hemolysis became evident. The DAT was still weakly positive and the serum contained a weak cold autoagglutinin, which did not correlate with the severity of the hemolysis. A Donath-Landsteiner test was performed and found to be strongly positive. The antibody showed P specificity, confirming a diagnosis of paroxysmal cold hemoglobinuria (PCH). Exchange transfusion was followed by rapid recovery even though the Donath-Landsteiner test remained positive for at least a month. The patient was well when last seen 11 months after presentation. It was thought that the original low titer of anti-A reflected compromised immune homeostasis in an elderly patient and that stimulation by incompatible blood in those circumstances resulted in a delayed hemolytic transfusion reaction that triggered, exacerbated, or was accompanied by an autoimmune response manifesting as PCH.
Immunohematology1997;13:5457.

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Case report: reporting anti-G as anti-C+D may have misleading clinical implications

A.L. Mosley, M.B. Trich, N.C. Thomas, and S.G. Sandler

Four months after a D male was transfused with four units of D red blood cells (RBCs), the results of a standard pretransfusion antibody screen and alloantibody identification panel detected anti-C+D in his serum. This report was interpreted by his physician to be evidence of alloimmunization to the D antigen, which triggered concern that the patient had been transfused previously with D+ RBCs as the result of an error in blood typing or personal identification. After a review of hospital records failed to identify such an error, consultation with a reference laboratory technologist confirmed that the serologic reactions resembled those of anti-C+D but were also consistent with anti-C + anti-G. Additional testing confirmed that the reactions were due to anti-G, not anti-C+D. One of the four donors was identified to have the C+D RBC phenotype, which is typically G+, thus identifying the stimulus for anti-G. Routine reporting of the detection of anti-C+D in the serum of D people, without confirmatory testing or commentary about the possibility that anti-G may resemble anti-C+D, may mislead health care providers who are not familiar with the pertinent blood group serology.
Immunohematology1997;13:5860.

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A case of hemolytic disease of the newborn due to anti-Kpa

L. Costamagna, M. Barbarini, G.L. Viarengo, A. Pagani, D. Isernia, and L. Salvaneschi

A patient with hemolytic disease of the newborn (HDN) due to maternal anti-Kpa alloimmunization is described. Although there are few reports in the literature, it appears that HDN due to anti-Kpa is often mild and transfusion therapy is rarely required. However, in this case, the baby's hemoglobin progressively decreased and on day 18 a blood transfusion was administered, but jaundice was not severe enough for exchange transfusion.
Immunohematology1997;13:6162.

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