Analysis strategies to establish vWF intron 40 haplotypes

Sequencing of a 0.65 kb region in the intron 40 of the vWF gene demonstrated a complex variability. Five STRs named Pol K, F, P1, P2-a and P2-b and an indel polymorphisms (I) are present. We established a routine analysing method to puzzle out the Pol K/F/I/P haplotypes which does not require a sequencing procedure. To recognise the combined polymorphisms as haplotypes, we performed short and middle range PCRs in combination with Nde I and BsmA I restriction tests. Comparison of the amplicon and restriction fragment length reveals the most likely haplotypes of each person involved a kinship test. Furthermore, a SNP allele specific PCR was employed. Additional information can be achieved by typing Pol P2-a and P2-b. Establishing of intron 40 vWF haplotypes using the methods described here can greatly support the resolution of complex kinship cases. This statement is illustrated by demonstration of a family study.