HPA genotyping by PCR sequence‐specific priming (PCR–SSP): a streamlined method for rapid routine investigations

Cavanagh, G.; Dunn, Augustine; Chapman, C.; Metcalfe, Paul

doi:10.1046/j.1365-3148.1997.d01-72.x

Cited by 96 publications

(91 citation statements)

References 10 publications

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“…12 HPA genotyping was performed using the polymerase chain reaction technique with sequence specific primers (PCR-SSP). 13 Clinical data were obtained from hospital records, as well as directly from the parents of affected infants or from their family doctors in some cases.…”

Section: Methodsmentioning

confidence: 99%

Neonatal alloimmune thrombocytopenia in the Irish population: a discrepancy between observed and expected cases

Davoren

McParland²,

Barnes³

et al. 2002

Journal of Clinical Pathology

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Aims: To estimate the rate of detection of neonatal alloimmune thrombocytopenia (NAITP) in the Irish population, to investigate clinical presentation and outcome in affected infants, and to determine the extent, if any, to which this condition is underdiagnosed at present. Methods: Cases were collected in a retrospective fashion from a review of platelet serology laboratory records from January 1992 to December 2000. Clinical data were obtained from hospital records. Testing for maternal antiplatelet antibody was by one or more of the following: the platelet suspension immunofluorescence test, a commercial antigen capture enzyme linked immunosorbent assay (GTI-PakPlus®), and the monoclonal antibody immobilisation of platelet antigens assay. Platelet antigen typing was by the polymerase chain reaction technique with sequence specific primers. Results: Twenty seven serologically verified cases of NAITP were identified in 18 families. Maternal antibody to human platelet antigen 1a accounted for 25 of the 27 confirmed cases. Twenty one of 26 infants were born with severe thrombocytopenia. Nineteen of 27 infants had bleeding manifestations at birth. Petechiae and bruising were most commonly observed (n = 17). There were no documented cases of intracranial haemorrhage in this group but systematic cranial ultrasound was not performed. Conclusions: Screening studies in predominantly white populations have estimated the incidence of NAITP to be between 1 in 1000 and 1 in 2000 live births. With 50 000 births each year in Ireland, these results give a clinical detection rate for NAITP of just 1 case in 16 500 live births, strongly suggesting that NAITP is currently underdiagnosed. Antenatal screening to detect women at risk of having babies with NAITP is now scientifically feasible and should be considered.

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Section: Methodsmentioning

confidence: 99%

Neonatal alloimmune thrombocytopenia in the Irish population: a discrepancy between observed and expected cases

Davoren

McParland²,

Barnes³

et al. 2002

Journal of Clinical Pathology

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“…Analysis of methyl-tetra-hydrofolate-reductase variant (MTHFRC677T), tumor necrosis factor-␣, and the apolipoprotein B (XbaI, EcoRI, and MspI) SNPs were performed by the Department of Haematology, Addenbrooke's Hospital, according to previously published methods. 18 HPA genotyping for human platelet antigen (HPA) 1, 2, 3, and 5 was performed using the PCR with sequence-specific priming (SSP) method described by Cavanagh et al 19 by the National Blood Service East Anglia Centre, Addenbrooke's Hospital.…”

Section: Determination Of the Origin (Donor Versus Recipient) Of Peptmentioning

confidence: 99%

Late diversification in the clonal composition of human cytomegalovirus-specific CD8+ T cells following allogeneic hemopoietic stem cell transplantation

Gandhi

Wills

Okecha

et al. 2003

Blood

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“…We used the HPA-1(24-45) peptide and also a shorter peptide with an extended N-terminal sequence (20)(21)(22)(23)(24)(25)(26)(27)(28)(29)(30)(31)(32)(33)(34)(35)(36)(37)(38)(39) in both the Leu33 (HPA-1a) and Pro33 (HPA-1b) forms in T cell proliferation assays to detect responding T cells from women who had been alloimmunized by pregnancy. Most samples were taken from patients during pregnancy or postpartum.…”

Section: Introductionmentioning

confidence: 99%

Reactivity of T cells from women with antibodies to the human platelet antigen (HPA)-1a to peptides encompassing the HPA-1 polymorphism

Jackson

Soothill

et al. 2005

Clinical and Experimental Immunology

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SummaryThe human platelet antigen-1a (HPA-1a) is the most common alloantigenic target in fetal and neonatal alloimmune thrombocytopenia (NAIT). Treatment currently depends on the outcome in previous pregnancies. HPA-1 specific T cell responses were determined in 14 HPA-1a alloimmunized women during or after pregnancies affected by NAIT. Peripheral blood mononuclear cells were incubated with peptides encompassing the Leu33Pro polymorphism (residues 20-39 and 24-45 in both Leu33 (HPA-1a) and Pro33 (HPA1b) forms) or control recall antigens in the presence of autologous sera and T cell proliferation was measured by 3 H-thymidine incorporation. Control antenatal and postpartum sera suppressed T cell proliferation and use of such sera was avoided. Most patients (86%) responded to the HPA-1a peptides with 64% also having weaker T cell proliferation to the HPA-1b peptides; 14% had no activity towards any peptide despite responding to control antigens. Administration of IVIG during pregnancy appeared to reduce T cell reactivity to HPA-1 peptides. Postnatal anti-HPA-1a T cell responses from women who had a severe history of NAIT (an intracranial haemorrhage in a previous fetus) were greater than those from women with a mild history. This assay may have the potential to predict disease severity if performed prior to or early in pregnancy.

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HPA genotyping by PCR sequence‐specific priming (PCR–SSP): a streamlined method for rapid routine investigations

Cited by 96 publications

References 10 publications

Neonatal alloimmune thrombocytopenia in the Irish population: a discrepancy between observed and expected cases

Neonatal alloimmune thrombocytopenia in the Irish population: a discrepancy between observed and expected cases

Late diversification in the clonal composition of human cytomegalovirus-specific CD8+ T cells following allogeneic hemopoietic stem cell transplantation

Reactivity of T cells from women with antibodies to the human platelet antigen (HPA)-1a to peptides encompassing the HPA-1 polymorphism

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