A total of 109 inversion negative HA (39 severe, 34 moderate and 36 mild) cases were included in this study of mutation screening by CSGE followed by DNA sequencing.Mutations could be detected in 98/109 (89.9%) cases. A total of 69 different mutations were identified of which 29 were novel and 40 recurrent. Of these unique mutations, 42 were missense, 8 nonsense, 13 deletions, 5 insertions and 1 splice site mutation. Double mutations were identified in 2 unrelated families. A classical case of female HA was also identified belonging to a family with strong history of moderate HA and consanguinity. An exon 14 polymorphism was identified with high heterozygosity frequency of 32.4%. No common mutation or founder mutation was identified in the present study.
82 inhibitor positive patients and 72 severe HA controls i.e. severe haemophilia A patients (> 10 years of age, who have had > 10 exposures to blood products or recombinant factors) were included in the study. The IL10 promoter polymorphisms 'GCC/ATA' ,HLA DRB1*15 and DQB1*05, DQB1*06, DRB1*03 ,Intron 22 Inversions , large deletions and missense mutations were significantly different between inhibitor positive and inhibitor negative patients.
112 subjects (150 including family members) were included in the study. 20 novel and 3 known candidate variations (mono-allelic) were detected in 48 subjects. . Different types of mutations were detected in different genes i.e. GP1BA (4), GP1BB (3), GP9 (9), ABCG5 (4) and MYH9 (3) genes which confirms the heterogeneity of this disorder. By Suppressive Subtractive Hybridization (SSH), 20 genes were found to be differentially expressed in patient's platelets; the expression profile of 4 genes Iron Responsive Element Binding protein 2 (IREB2), Alpha Tubulin (TUBA), Myotubularin related protein 9 (MTMR9) and Tyrosine Kinase (TKL) of the 20 genes was further confirmed by mRNA expression level by quantitative RT-PCR. The study revealed that new genes might be involved in the pathogenesis of the disorder which needs to be confirmed by further studies.
Haemorrhagic disease of the newborn (HDN)or delayed vitamin K deficiency related bleeding is an uncommon disorder with potentially devastating outcome.The bleeding is often intracranial, bot also can be mucosal or gastrointestinal . 40 infants with HDN were analyzed for various polymorphisms in the vitamin K and gamma carboxylation pathway.. More than half of the children had warfarin sensitive alleles indicating that the same polymorphisms may have a role in their manifestation.
Two workshops and three on spot training programmes were conducted on laboratory diagnosis of bleeding disorders. More than 100 laboratory technicians and Pathologists from different states in Central and North eastern India were trained in basic coagulation techniques.
Approximately 4-11 % of the patients with idiopathic venous thrombosis (VT) show protein C (PC) deficiency. The molecular pathology of PC deficiency was analyzed in 102 patients ; 98 healthy controls were also studied to assess the association of various polymorphisms with reduced PC levels. PROC gene mutations were detected only in 8 (7.8) % patients with reduced PC levels. PROC promoter region CG polymorphisms showed statistically significant association with reduced PC levels (p 0.001). PC deficiency in Indian VT patients can thus largely be explained by PROC gene promoter CG polymorphisms; only a small fraction of the patients show specific mutations in PROC gene.
747 patients with thrombosis at different sites were screened for the presence of JAK2 mutations. 48 patients (6.42%) were found to be carriers of JAK2 mutation. Highest prevalence of JAK2 was found in BCS (13.5%), followed by MVT (8.3%) and PVT (7.1%) cases. Among the CVT and DVT cases, the prevalence was found to be 4.5% and 1.6% respectively. None of the controls screened were positive for JAK2 mutation.
131 type 3 VWD patients from 100 VWD families (100 unrelated severe index cases) were analysed in the present study. Mutations in 8 different CGA codons were identified in 27 patients (unrelated 20 patients). Thus, we could detect 20 homozygous (one each in exon 3, 28 and 45, three in exon 8, four in exon 9, and five in exon 31, 43) and 7 (one in exon 45 and two in exon 10 and four in 31) heterozygous mutations by this relatively simple technique . Different types of mutations were detected by CSGE based on mobility shift and sequencing i.e., missense, deletion, duplication, nonsense and frameshift mutation could be detected by PCR-CSGE followed by purification and sequencing. Our study could characterize mutation in 77% cases (89 type 3 VWD patients). 21 novel mutations were detected and 16 different mutations were identified in propeptide region in 20 patients. Thus this study in large cohort of Indian type 3 VWD patients adds further insights in mechanisms associated with VWD.
A simple and cost effective two-tube technique was developed for the detection and differentiation of severe and mild-to-moderate FXIII deficiency.F13A1 mutations in 44 severe FXIII deficient patients have been identified.High heterogeneity was observed in this study with 19 novel mutations.
In the present series, there were 7 cases of afibrinogenemia, 8 hypofibrinogenaemia and 4 dysfibrinogenaemia.14 mutations were identified in the present study in 16 patients, out of which 10 were novel and 4 reported. 5 mutations were in FGA, 5 in FGB and 4 were in FGG. 6 were missense, 2 nonsense, 3 deletions, 1 insertion, 1 insertion-deletion and 1 splice site mutation.2 families were referred for antenatal diagnosis during the first trimester of pregnancy and both the fetuses were found to be unaffected.