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Biomedical subjects

A C Goodeve

Publications and source records attributed to A C Goodeve.

29 records · Page 2Linked to original sources
PubMed1998

Advances in carrier detection in haemophilia.

Increasing worldwide use of molecular genetic analysis is enabling accurate carrier detection for the haemophilias to be made more widely available. Use of DNA polymorphisms in linkage analysis is an accurate method for carrier detection applicable to the majority of families. For those families with severe haemophilia A, the inversion mutation can be sought by most molecular genetics laboratories. For families remaining uninformative by these procedures, a range of point mutation screening techniques is available. Dedicated electrophoresis equipment is enabling use of these techniques to become more widespread.

Alleles↗
PubMed1998

Precise carrier diagnosis in families with haemophilia A: use of conformation sensitive gel electrophoresis for mutation screening and polymorphism analysis.

Causative mutations in the factor VIII gene of seven unrelated patients with severe haemophilia A were identified using the mutation screening procedure conformation sensitive gel electrophoresis (1) and characterised by direct sequencing. Female family members of all patients had requested either carrier status determination or prenatal diagnosis. However, lack of the factor VIII gene inversion, a prior family history or informative polymorphisms prevented diagnosis in these families. Identification of a mutation in each family enabled female carrier status to be determined in all cases. Six mutations were previously unreported. One Afro-Caribbean patient had two sequence changes; A670 2G and A6769G. The latter, resulting in Met2238Val and previously reported as a FVIII mutation, was shown to be polymorphic with a 42% heterozygosity rate in an Afro-Caribbean population. Conformation sensitive gel electrophoresis was found to be technically simple and efficient at locating previously unknown FVIII gene mutations.

Blotting, Southern↗
PubMed1997

HLA class II profile: a weak determinant of factor VIII inhibitor development in severe haemophilia A. UKHCDO Inhibitor Working Party.

The risk of developing factor VIII inhibitor antibodies in haemophilia A may relate both to factor VIII genotype and genes within the HLA complex known to influence immune response. We investigated a cohort of 176 patients with severe haemophilia A and with either high-level inhibitors (> 10BU/ml) or with no history of an inhibitor, stratified according to the presence or absence of the factor VIII gene intron 22 inversion. HLA DRB1, DQA1 and DQB1 polymorphisms were determined by PCR. HLA frequencies form 137 United Kingdom controls were used for comparison. HLA phenotype frequency differences, expressed as odds ratios with 95% confidence intervals were as follows: HLA-DRB*1501, DQB1*0602 and DQA1*0102 were all increased in frequency in patients with inhibitors, only DQA1*0102 reaching statistical significance (OR 2.7, 1.2-5.9). These alleles form part of an established HLA haplotype. The frequencies of HLA-DRB1*1501, DQB1*0602 and DQA1*0102 were particularly raised in patients with inhibitors and a factor VIII gene intron 22 inversion, although again only DQA1*0102 achieved significance (OR 3.1, 1.0-10.1). The frequency of DRB1*01, DQB1*0501, DQA1*0101 were also increased in inhibitor patients lacking the intron 22 inversion although this failed to achieve statistical significance. This data suggests that HLA class II profile constitutes a weak risk factor for developing inhibitor antibodies to factor VIII. This may be more pronounced in patients with an intron 22 inversion.

Chromosome Inversion↗
PubMed1996

A rapid and cost effective method for analysis of dinucleotide repeat polymorphisms in the factor VIII gene.

A simple, rapid and cheap method for the analysis of variable dinucleotide tandem polymorphisms in introns 13 and 22 of the factor VIII gene has been established. The protocol uses blood spots stored on filter paper (Guthrie spots) and other DNA containing materials as sources of DNA. Following DNA amplification using a thermostable DNA polymerase, products are size fractionated on native polyacrylamide gels and visualized by silver staining. This simplified protocol obviates the use of DNA extraction, reducing time and costs and in addition, reduces the requirement for gel documentation equipment (i.e., photography), as silver staining can be visualized readily without additional equipment and the gels themselves can be stored. These alterations to the analysis enhance the universal applicability of these polymorphisms, as was demonstrated by a comparative study in Caucasian and Thai females.

Cost-Benefit Analysis↗
PubMed1996

Characterisation of type 2N von Willebrand disease using phenotypic and molecular techniques.

von Willebrand factor (vWF) is a multimeric glycoprotein found in plasma non covalently linked to factor VIII (FVIII). Type 2N von Willebrand disease (vWD) is caused by a mutation in the vWF gene that results in vWF with a normal multimeric pattern, but with reduced binding to FVIII. We have utilised methods for the phenotypic and genotypic detection of type 2N vWD. The binding of FVIII to vWF in 69 patients, 36 with type 1 vWD, 32 with mild haemophilia A and one possible haemophilia A carrier with low FVIII levels was studied. Of these, six were found to have reduced binding (five type 1 vWD, one possible haemophilia A carrier). DNA was extracted from these patients and exons 18-23 of the vWF gene encoding the FVIII binding region of vWF were analysed. After direct sequencing and chemical cleavage mismatch detection, a Thr28Met mutation was detected in two unrelated individuals, one of whom appears to be a compound heterozygote for the mutation and a null allele. No mutations were found in the region of the vWF gene encoding the FVIII binding region of vWF in the other four patients.

Alleles↗
PubMed1994

Factor VIII gene rearrangements in patients with severe haemophilia A.

Sequences within intron 22 of the factor VIII (FVIII) gene have been implicated in the cause of haemophilia in almost 50% of severely affected patients. The changes result from intrachromosomal rearrangements of the tip of the long arm of the X chromosome, one break-point being within intron 22 of the FVIII gene. The rearrangements can be identified by Southern blot and we report use of this procedure to detect rearrangements in 11 of 23 unrelated families with severe haemophilia A. Of 22 patients studied, none of the 10 with the gene rearrangement had at any time developed inhibitors to FVIII, compared with 7 of 12 lacking the rearrangement.

Adult↗
PubMed1994

A comparison of the allelic frequencies of ten DNA polymorphisms associated with factor VIII and factor IX genes in Thai and Western European populations.

The frequency of five factor VIII gene intragenic and linked DNA polymorphisms and five factor IX gene intragenic polymorphisms was studied in Thai females. The polymorphisms in the FVIII gene were detected by restriction enzymes BclI, XbaI, BglI and at linked loci DX13 (DXS15) and St14 (DXS52) by BglII and TaqI, respectively, and in the FIX gene by MseI, DdeI, XmnI, TaqI and HhaI. With the exception of the BglI restriction fragment length polymorphism (RFLP), which is absent in Thais, factor VIII polymorphism frequencies were similar in Thais and Caucasians. Combined use of XbaI and TaqI/St14 resulted in a heterozygosity rate of greater than 90% in Thai females. For FIX, the recently described MseI RFLP in the 5' flanking region was the most informative polymorphism in Thais, 43% of females being heterozygous. The other four polymorphisms added little to the overall heterozygosity rate. The appropriate polymorphisms were used to track defective factor VIII and IX genes through 22 Thai pedigrees with haemophilia to enable carrier status to be assigned to female family members. The information obtained during this study will form the basis for carrier detection and prenatal diagnosis of haemophilia A and B by DNA polymorphism analysis in Thailand.

Alleles↗
PubMed1992

Detection of EBV DNA in post-nasal space biopsy tissue from asymptomatic EBV-seropositive individuals.

The association between EBV and nasopharyngeal carcinoma (NPC) has been well documented although the precise role of the virus in the genesis of the tumour is not understood. We undertook this study to examine the prevalence of EBV infection in nasopharyngeal tissue obtained from 33 healthy individuals not considered to be at risk of developing NPC. Using polymerase chain amplification (PCR) and in situ hybridization we have identified EBV DNA in 70% (23/33) of the tissues examined. Our data demonstrate that EBV is present at the site of tumour development in the low-risk population and by inference that the virus is also present before the onset of disease in the high-risk group. This survey supports the concept of NPC pathogenesis as a multifactorial process.

Base Sequence↗
PubMed1985

Reassortants of influenza B viruses for use in vaccines: an evaluation.

Three slightly different procedures for the preparation of influenza B virus reassortants from B/Lee/40 and B/Lyon/79, and B/Johannesburg/58 and B/Hong Kong/82 parental viruses are described. Following cloning procedures in eggs or allantois-on-shell cultures in the presence of antisera to B/Lee or B/Johannesburg, twenty-eight putative reassortants were obtained. Using SDS-PAGE to determine the migration rates of virion polypeptides, eighteen isolates from mixed infections of B/Lee with B/Lyon, and three isolates from mixed infections of B/Johannesburg with B/Hong Kong, were found to be reassortants. All reassortants possessed surface proteins derived from either B/Lyon or B/Hong Kong and one to three internal polypeptides from either B/Lee or B/Johannesburg. None of the reassortants showed a growth capacity in embryonated eggs as high as that of their B/Lee or B/Johannesburg parent viruses. The procedures used, and the influenza B reassortants thus generated, would appear to offer no major advantages over the existing methods and influenza B strains at present employed for the preparation of inactivated influenza virus vaccines.

Animals↗
PubMed1983

The use of the single radial haemolysis test for assessing antibody response and protective antibody levels in an influenza B vaccine study.

Single radial haemolysis (SRH) and HAI tests were used to determine the levels of antibody in sera obtained from 118 volunteers taking part in a study of influenza B/Hong Kong/73 subunit vaccine. The two tests gave similar overall results, and showed good correlation, but SRH appeared to be more sensitive for the detection of low concentrations of antibody. A greater than or equal to 45% increase in SRH zone area was found to be equivalent to a significant increase in antibody titre and on the basis of this value, the HAI and SRH tests gave similar results for responses to immunization. The concentration of antibody equivalent to a 50% protective level against a challenge virus infection was a zone area of 45 mm2 for the SRH test; this was equivalent to an HAI titre of 1:20 which has previously been established as the 50% protective level.

Antibodies, Viral↗