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Fenotypning av trombocytantigen HPA1a med flödescytometri: screening för att finna blodgivare som saknar HPA1a
Jönköping University, School of Health and Welfare, HHJ, Dep. of Natural Science and Biomedicine.
Jönköping University, School of Health and Welfare, HHJ, Dep. of Natural Science and Biomedicine.
2018 (Swedish)Independent thesis Basic level (degree of Bachelor), 10 credits / 15 HE creditsStudent thesis
Abstract [sv]

HPA1a är ett antigen på trombocytytan som kan orsaka alloimmunisering, såsom neonatal alloimmun trombocytopeni och post-transfusion purpura, vilket kan ge svåra blödningssymptom. I fall med antikroppar mot HPA1a måste kompatibla trombocyter finnas tillgängliga. Syftet med studien var att etablera en flödescytometrisk screeningmetod för fenotypning av HPA1a antigen på trombocyter samt att finna HPA1a negativa blodgivare. Före flödescytometrisk analys poolades två till fem blodprover samman till ett prov och vid förekomst av HPA1a negativa trombocyter i poolen analyserades proverna individuellt. Fluorokrommärkta anti-humana antikroppar mot CD42a och CD61 användes för att särskilja HPA1a negativa trombocyter från HPA1a positiva. Totalt fenotypades 177 blodprover varav sju (4%) typades som HPA1a negativa. Av de sju fynden genotypades fyra vid ett externt laboratorium vilket bekräftade att de var HPA1a negativa. Flödescytometrisk screening av HPA1a är snabb, pålitlig och lämplig för storskalig screening. För att fastställa prevalensen av HPA1a negativa individer behöver mer omfattande studier utföras där en större population ingår. Att ha flera HPA1a negativa blodgivare registrerade ger möjligheten att hjälpa patienter i andra regioner.

Abstract [en]

HPA1a is an antigen on the platelet surface that can cause alloimmunization, such as neonatal alloimmune thrombocytopenia and post transfusion purpura, which can cause severe bleeding symptoms. In case of antibodies against HPA1a, compatible platelets must be available. The purpose of the study was to establish a flow cytometric screening method for phenotyping HPA1a antigen on platelets and to find HPA1a negative donors. Before the flow cytometric analysis, two to five blood samples were pooled into one sample and in the presence of HPA1a negative platelets in the pool, the samples were analyzed individually. Fluorochrome –labeled anti-human antibodies to CD42a and CD61 were used to distinguish HPA1a negative platelets from HPA1a positive. A total of 177 blood samples were phenotyped, of which 7 (4%) were HPA1a negative. Of the seven findings, four samples were genotyped at an external laboratory confirming that they were HPA1a negative. Flow cytometric screening of HPA1a is fast, reliable and suitable for large scale screening. In order to determine the prevalence of HPA1a negative individuals, more extensive studies need to be performed involving a larger population. By having many registered HPA1a negative donors, it can provide opportunities to help patients in other regions.

Place, publisher, year, edition, pages
2018.
Keywords [en]
HPA1b, GPIIIa, thrombocytopenia, antibodies, alloimmunization
Keywords [sv]
HPA1b, GPIIIa, trombocytopeni, antikroppar, alloimmunisering
National Category
Biomedical Laboratory Science/Technology
Identifiers
URN: urn:nbn:se:hj:diva-40074ISRN: JU-HHJ-BLA-1-20180070OAI: oai:DiVA.org:hj-40074DiVA, id: diva2:1215989
Subject / course
HHJ, Biomedical Laboratory Science
Supervisors
Examiners
Available from: 2018-06-21 Created: 2018-06-10 Last updated: 2018-06-21Bibliographically approved

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