there is reseacrh paper regarding pcr diagnosis of hemophilia

Thank you for your help.

Is there any much simplified version of PCR for confirmation of hemophilia?

Hi Hiasindh Ashmi,

actually I do not think that you will find kits available for hemophilia PCR covering for example the 10 most common SNPs...most coagulation factor genes are rather large, and often there is more than one mutation on different parts of the gene, quite far apart from each other... Therefore, "simple" methods like SSP or SSO with only a few primers will not give satisfying results. You will probably have to establish a big panel of primers.

As NGS is getting more commen these days, maybe you could find some people having started projects on diagnosing hemophilia by NGS techniques.

What do you need the PCR for, if I may ask?

Thank you for your help Julian Strobel.

Recently, one of my friend has been diagnosed with recessive for bleeding disorder, with factor 8 deficiency. No clinical family history also. It was done by coagulation test.

I want to confirm it by molecular test. So only asked. I have extracted the DNA. If you know anything regarding this, please help me out.

Thanks in advance.

Hi,

we do not do hemophilia diagnosis by PCR methods, we use coagulation tests as well. Was the coagulation test repeated (at least twice)? There is no von Willebrand syndrome? No aquired hemophilia, may be due to inhibitors?

Sorry I cannot help with molecular tests!

Ya...coagulation test was repeated twice.. factor 8 deficiency is there and test has been done for von Willebrand syndraome using ELISA also on research basis by PhD scholar. It has come positive. But dont know how to interpret the result.

Can you tell me whether factor 8 deficiency and von Willebrand syndrome is linked? Also, whats the standard method to diagnose von Willebrand syndrome?

Thanks in advance.

Hi,

we normally do coagulation tests for von Willebrand´s disease as well. To be honest, we normally do a set of tests in case of bleeding tendency containing activities of FVIII, FIX, FXIII, vWF. Furthermore, we meassure the concentrations of fibrinogen and VWF-antigen. Finally, we do aPTT, prothrombin time and at least one test for platelet function, either PFA200 or Born´s aggregation test.

For vWD, we do platelet aggregation, FVIII, VWF antigen and activity; in some cases, we even do a vWF-multimer analysis.

Regarding the link of vWF and FVIII: Without vWF activity, you won´t have FVIII activity either, as vWF "protects" FVIII so the FVIII is not cleared as easily in plasma. Type 3 von Willebrand´s disease (complete missing of vWF) therefore presents bleeding symptoms like hemophilia A. By the way: Your friend is male or female?

My friend is a female and she has complaints of recurrent ecchymosis for the past two years. As she complained, we did a screening test PT, APTT, TT and Fibrinogen. Then as we suspect platelet function disorder and vWD, platelet function test was done. This test showed defective aggregation pattern with ristocetin. This suggests presence of vWD. So for confirmation, we need to do vWF antigen assay and RiCoF. As we have a premature lab, what are the other investigation need to be done in a developing lab