MEDICINE
Year : 2010  |  Volume : 1  |  Issue : 1  |  Page : 25-28

HindIII-based restriction fragment length polymorphism in hemophilic and nonhemophilic patients


1 Division of Cellulose and Paper, Forest Research Institute, P.O., IPE, Kaulagarh Road, Dehradun, Uttarakhand, India
2 Department of Pathology, King George Medical University, Lucknow, Uttar Pradesh, India
3 Division of Genetics and Tree Propagation, Forest Research Institute, P.O., IPE, Kaulagarh Road, Dehradun, Uttarakhand, India

Correspondence Address:
Alok Kumar Dubey
Cellulose and Paper Division, Forest Research Institute, P.O., IPE, Kaulagarh Road, Dehradun - 248 195, Uttarakhand
India
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/0976-9668.71669

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Hemophilia A is most common recessively inherited bleeding disorder, which affect one in five thousand male births throughout the world. In most of the hemophilic A patients, no common mutation is easily identifiable. This limitation has been overcome by the use of polymorphic DNA marker, i.e., restriction fragment length polymorphism (RFLP). This marker of polymorphism could only be detected by amplifying the polymorphic region and digestion the polymerase chain reaction (PCR) product with the restriction enzyme (PCR−RFLP), i.e., HindIII. The polymorphic region of HindIII is 608 bp in length and after the restriction digestion, different sizes of fragments, i.e., 427 and 181 bp were, respectively, obtained. However, in homozygous (+/+) condition three bands of 427, 100, and 81 bp were obtained and in the other negative allelic homozygous condition (-/-) two bands of 427 and 181 bp were obtained. Similarly fragments of different sizes, i.e., 427, 181, 100, and 81 bp were obtained in heterozygous conditions. Therefore, in this study, we have analyzed the factor VIII gene in the 17 different families using restriction enzyme HindIII-based RFLP molecular marker technique. Out of these, the observed heterozygosity for HindIII was found 47.5%, whereas, for positive allele it was 26%, and for negative allele the frequency was 74%.


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