Carrier detection and prenatal diagnosis by intron 22 inversion analysis of the factor VIII gene

SummaryHaemophilia A (HA) is X-chromosome linked bleeding disorders caused by deficiency of the coagulation factor VIII (FVIII). It is caused by FVIII gene intron 22 inversion (Inv22) in approximately 45% and by intron 1 inversion (Inv1) in 5% of the patients. Both inversions occur as a result of intrachromosomal recombination between homologous regions, in intron 1 or 22 and their extragenic copy located telomeric to the FVIII gene. The aim of this study was to analyze the presence of these mutations in 25 HA Costa Rican families. Patients, methods: We studied 34 HA patients and 110 unrelated obligate members and possible carriers for the presence of Inv22or Inv1. Standard analyses of the factor VIII gene were used incl. Southern blot and long-range polymerase chain reaction for inversion analysis. Results: We found altered Inv22 restriction profiles in 21 patients and 37 carriers. It was found type 1 and type 2 of the inversion of Inv22. During the screening for Inv1 among the HA patient, who were Inv22 negative, we did not found this mutation. Discussion: Our data highlight the importance of the analysis of Inv22 for their association with development of inhibitors in the HA patients and we are continuous searching of Inv1 mutation. This knowledge represents a step for genetic counseling and prevention of the inhibitor development.

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Inversions in the factor VIII gene: improvement of carrier detection and prenatal diagnosis in Dutch haemophilia A families.

Journal of Medical Genetics ◽

10.1136/jmg.32.4.296 ◽

1995 ◽

Vol 32 (4) ◽

pp. 296-300 ◽

Cited By ~ 3

Author(s):

P P Deutz-Terlouw ◽

M Losekoot ◽

R Olmer ◽

W C Pieneman ◽

S de Vries-v d Weerd ◽

...

Keyword(s):

Prenatal Diagnosis ◽

Factor Viii ◽

Carrier Detection ◽

Haemophilia A ◽

Factor Viii Gene

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Factor VIII gene inversions and polymorphisms in Brazilian patients with haemophilia A: carrier detection and prenatal diagnosis

Haemophilia ◽

10.1046/j.1365-2516.2001.00508.x ◽

2001 ◽

Vol 7 (3) ◽

pp. 299-305 ◽

Cited By ~ 20

Author(s):

R. P. S. Soares ◽

D. A. F. Chamone ◽

S. P. Bydlowski

Keyword(s):

Prenatal Diagnosis ◽

Factor Viii ◽

Carrier Detection ◽

Haemophilia A ◽

Factor Viii Gene

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Carrier detection in haemophilia A by direct analysis of factor VIII gene lesions

Human Genetics ◽

10.1007/bf01213104 ◽

1991 ◽

Vol 87 (1) ◽

pp. 99-100

Author(s):

David S. Millar ◽

Peter J. Green ◽

Barbara Zoll ◽

Vijay V. Kakkar ◽

David N. Cooper

Keyword(s):

Factor Viii ◽

Direct Analysis ◽

Carrier Detection ◽

Haemophilia A ◽

Factor Viii Gene

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Analysis of intron 22 inversions of the factor VIII gene in severe hemophilia A: implications for genetic counseling

Blood ◽

10.1182/blood.v84.7.2197.2197 ◽

1994 ◽

Vol 84 (7) ◽

pp. 2197-2201 ◽

Cited By ~ 33

Author(s):

PV Jenkins ◽

PW Collins ◽

E Goldman ◽

A McCraw ◽

A Riddell ◽

...

Keyword(s):

Factor Viii ◽

Genetic Counselling ◽

Hemophilia A ◽

Family Members ◽

Rflp Analysis ◽

Homologous Region ◽

Severe Hemophilia ◽

Factor Viii Gene ◽

Intron 22 Inversion ◽

Blotting Technique

Abstract Intrachromosomal recombinations involving F8A, in intron 22 of the factor VIII gene, and one of two homologous regions 500 kb 5′ of the factor VIII gene result in large inversions of DNA at the tip of the X chromosome. The gene is disrupted, causing severe hemophilia A. Two inversions are possible, distal and proximal, depending on which homologous region is involved in the recombination event. A simple Southern blotting technique was used to identify patients and carriers of these inversions. In a group of 85 severe hemophilia A patients, 47% had an inversion, of which 80% were of the distal type. There was no association with restriction fragment length polymorphism (RFLP) haplotypes. The technique has identified a definitive genetic marker in families previously uninformative on RFLP analysis and provided valuable information for genetic counselling information may now be provided for carriers without the need to study intervening family members and the diagnosis of severe hemophilia A made in families with only a nonspecific history of bleeding. Analysis of intron 22 inversion should now be the first-line test for carrier diagnosis and genetic counselling for severe hemophilia A and may be particularly useful when there is no affected male family member or when intervening family members are unavailable for testing.

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Genetic Counseling by Analysis of Intron 22 Inversions of the Factor VIII Gene

Clinical and Applied Thrombosis/Hemostasis ◽

10.1177/107602969800400206 ◽

1998 ◽

Vol 4 (2) ◽

pp. 111-113

Author(s):

Etsuko Yamazaki ◽

Hiroshi Mohri ◽

Hiroshi Harano ◽

Heiwa Kanamori ◽

Hiroshi Inaba ◽

...

Keyword(s):

At Risk ◽

Genetic Counseling ◽

Factor Viii ◽

Key Words ◽

Gene Rearrangement ◽

Hemophilia A ◽

Carrier Detection ◽

Severe Hemophilia ◽

Normal Allele ◽

Factor Viii Gene

Ten patients with severe hemophilia A and 10 with moderate and mild hemophilia A were studied. Five of 10 unrelated patients with severe hemophilia A had the distal telo meric int22h sequence, none had the proximal sequence, and one had a unique variant factor VIII gene rearrangement. Car rier detection was done in these six families. All mothers and two daughters of the patients were to be carriers. Six of the 15 at-risk female relatives were heterozygous for the rearranged and normal allele and were carriers. These results indicate that the rearrangement assay is very useful for carrier detection in families with severe hemophilia A. Key Words: Hemophilia A—Factor VIII gene rearrangement—Genetic counseling.

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PRENATAL DIAGNOSIS OF HAEMOPHILIA A BY FACTOR VIII GENE ANALYSIS

The Lancet ◽

10.1016/s0140-6736(85)91842-2 ◽

1985 ◽

Vol 325 (8443) ◽

pp. 1407-1409 ◽

Cited By ~ 35

Author(s):

StylianosE Antonarakis ◽

KarenL Copeland ◽

RobertJ Carpenter ◽

CarlA Carta ◽

LeonW Hoyer ◽

...

Keyword(s):

Prenatal Diagnosis ◽

Factor Viii ◽

Gene Analysis ◽

Haemophilia A ◽

Factor Viii Gene

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Haemophilia A: Mutation Type Determines Risk of Inhibitor Formation

Thrombosis and Haemostasis ◽

10.1055/s-0038-1649954 ◽

1995 ◽

Vol 74 (06) ◽

pp. 1402-1406 ◽

Cited By ~ 227

Author(s):

R Schwaab ◽

H-H Brackmann ◽

C Meyer ◽

J Seehafer ◽

M Kirchgesser ◽

...

Keyword(s):

Factor Viii ◽

Antibody Formation ◽

Gene Mutations ◽

Therapeutic Effects ◽

Haemophilia A ◽

Missense Mutations ◽

Factor Viii Gene ◽

Large Deletions ◽

Intron 22 Inversion ◽

First Time

SummaryThe formation of factor VIII antibodies is a major problem for replacement therapy of haemophilia A patients. Antibodies occur in 5-30% of patients with severe haemophilia A. The reason for antibody formation is still unknown. In this study we correlate for the first time different factor VIII gene mutations, stop- and missense mutations, large and small deletions and intrachromosomal intron 22 recombinations to antibody formation. A total of 364 patients with known inhibitor status of our institute, of the database, and of 3 studies representing intron-22-inversion data are included. The results show that the risk for developing factor VIII antibodies is strongly related to stop mutations, large deletions and intrachromosomal recombinations. A probable explanation could be the complete lack of endogenous circulating factor VIII protein in these cases. Other factors that might be important for the pathogenesis of inhibitor formation, e. g. the antenatal period, as well as possible therapeutic effects, are discussed.

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Another Variant Pattern of Intron 22 Inversion in the Factor VIII Gene Seen in a Severe Haemophilia A Patient

Thrombosis and Haemostasis ◽

10.1055/s-0038-1657738 ◽

1997 ◽

Vol 78 (04) ◽

pp. 1303-1303 ◽

Cited By ~ 2

Author(s):

Mohammed S Enayat ◽

Bimal D M Theophilus ◽

Michael D Williams ◽

Jonathan T Wilde ◽

Frank G H Hill

Keyword(s):

Factor Viii ◽

Haemophilia A ◽

Severe Haemophilia ◽

Factor Viii Gene ◽

Intron 22 Inversion ◽

Variant Pattern

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CARRIER DETECTION IN JAPANESE FAMILIES WITH HAEMOPHILIA A USING FACTOR VIII GENE PROBE(F8A) AND ST 14-1 PROBE

10.1055/s-0038-1644009 ◽

1987 ◽

Author(s):

M Nishino ◽

T Nishimura ◽

H Naka ◽

S Mikami ◽

A Yoshioka ◽

...

Keyword(s):

Prenatal Diagnosis ◽

Dna Probe ◽

Carrier Detection ◽

Carrier Status ◽

Gene Probe ◽

Haemophilia A ◽

Factor Viii Gene ◽

X Chromosomes ◽

The Family ◽

The Relationship

Recently, the gene structure for human F.VIII protein was clarified, and F.VIII DNA probes have been used for carrier detection and prenatal diagnosis ofhaemophilia A. In order to make sure that the phenomena are universal, we have analysed the RFLPs of F.VIII gene in 16 Japanese families with haemophilia A, including a female haemophiliac case, using an intragenic F.VIII DNA probe(F8A) and an extragenic(linked) DNA probe(Stl4-1).The probe F8A revealed two variant bands after digestion by Bel I. Of normal 60 X chromosomes (females) examined, about 85% bore the 879-bp fragment and 15%the 1165-bp fragment. Five of sixteen mothers of hemophiliacs, definite carriers, were found to be heterozygous for Bel I polymorphism. Since the relationship between Bel I alleles and hemophilia gene has been identified in the 5 families in which the mothers were heterozygous, we could diagnose the carrier status of two women whose brothers are hemophiliacs. Onthe other hand, we could identify that one "haemophilic woman" with less than 10% of F.VIII:C was a carrier status when we analysed the Bel I alleles in theother members of the family.The probe DNA(ST 14-1) revealed seven variant bands ranging from 5.5 kb to 3.4 kb after digestion by Taq I. In 6 out of 16 families, the RFLPs of ST 14 locus were informative for carrier detection.From these data, it was concluded that the Bel I polymorphism of F.VIII gene and the Taq I polymorphism of ST 14 locus were informative for carrier detection in 8 out of 16 families with haemophilia A

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