Carrier detection and prenatal diagnosis of haemophilia / 中国输血杂志

ABSTRACT

Objects To establish a simple,rapid carrier detection and prenatal diagnosis system for hemophilia.Methods Thirty-eight HA families were tested for the intron 22 and 1 inversions in factor VIII gene by LD-PCR and PCR.The remaining inversion negative families,but with family history,were screened using linkage analysis with 8 combined polymorphic markers,including St14,F8IVS13,CA22,DXS15,DXS9901,G6PD,DXS1073,and DXS1108.For sporadic families,the whole gene sequencing was applied directly to detect the mutation.For HB families,linkage analysis with 6 STRs,including DXS1192,DXS1211,DXS8094,DXS8013,DXS1227 and DXS102,was applied to get quick diagnostic information.The whole gene sequencing was used to get the final diagnosis.The rapid fluorescent PCR combined with polymorphism markers were applied for linkage analysis in HA and HB families,respectively.As soon as the pregnancy was identified,additional Amelo site detection would be performed.Results In 38 HA families,introns 22 and 1 inversions were found in 10 and 1 probands,respectively.The diagnostic rates of St14,F8IVS13,CA22,DXS15,DXS9901,G6PD,DXS1073 and DXS1108 were 61.11%,76.67%,71.43%,70.59%,62.50%,10.00%,75.00% and 50.00%,respectively.Combining inversion detection and linkage analysis,the diagnostic rate of carrier detection and prenatal diagnosis of families with HA family history were both 100%.One intron 22 inversion and 3 mutations were detected in 4 sporadic families.The total diagnostic rate of 38 HA families was 94.81%.And 10 mutations were detected in the 12 HB families.Combined with the linkage analysis,the total diagnostic rate was 96.88%.Conclusions Introns 22 and 1 inversion screening combined with the linkage analysis,using the highly informative polymorphic markers,can be used for carrier detection and prenatal diagnosis in Chinese HA families.The direct sequencing of FⅨ with the linkage analysis can be successfully applied for carrier detection and prenatal diagnosis of HB families.