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Showing 7 results for Karimipoor

Esmat Kamali Dolatabadi, Morteza Karimipoor, Shahram Samiee, Leila Kokabee, Siroos Zinali, Nafiseh Nafissi, Hamid Reza Hoorfar,
Volume 3, Issue 4 (Winter 2007)
Abstract

  Abstract

 Background and Objectives

 Hemophilia B is an inherited recessive X-linked bleeding disorder caused by deficiency or defect of procoagulant factor IX (FIX). The factor IX gene spans 35kb of DNA and comprises of 8 exons. Mutations in the factor IX gene may result in deficient or defective coagulation factor IX causing the bleeding tendency known as hemophilia B. The aim of this study was to identify the causative mutations and genotype-phenotype correlation for mutations in some known patients with hemophilia B in Isfahan province.

 

 Materials and Methods

 After informed consent was obtained, genomic DNAs of 24 hemophilia B patients referred to Omid hospital were extracted according to standard protocols. PCR amplification and single strand conformation polymorphism (SSCP) on nondenaturing polyacrylamid gel were performed separately on each sample for eight exons and exon-intron boundaries and promoter. The results of SSCP were compared to normal control and sequencing was performed for those with different migration patterns.

 

 Results

 The sequencing results showed 70.8% missense mutation, 16.7% deletion, 8.3% nonsense mutation, and 4.2% insertion. Many of the mutations had occurred in exon 8 it came out to be similar to haemophilia B mutation database. Malmo polymorphism (Ala 148 Thr) was found in one family. Four novel mutations not previously reported in the database were also found.

 

 Conclusions

 This study confirms the marked heterogeneity of factor IX mutations in the population. The results could be used to develop a national database and offer genetic counselling to families.

 

 Key words : Hemophilia B, SSCP, Mutation

 


S. Babashah, S. Jamali, Dr R. Mahdian, M. Hayat Nosaeid, Dr. M. Karimipoor, Dr. F. Maryami, M. Raeisi, R. Alimohammadi, Dr. S. Zeinali,
Volume 5, Issue 4 (Winter 2009)
Abstract

  Abstract

 Background and Objectives

 Although beta thalassemia is mainly caused by mutations involving single base substitutions and small deletions, there have been reports showing deletions of large regions of beta- globin genes play a similar causing role. The strategy to identify beta thalassemia carriers with known deletions is based on PCR techniques such as Gap PCR. There are however some unknown deletions that can not be detected by the above methods. To overcome this limitation, Real-time PCR and MLPA were developed as two quantitative assays for analysis of beta-globin gene cluster.

 

 Materials and Methods

 The subjects were evaluated in a case-control study. Among individuals referred to genetic laboratories of Pasteur Institute of Iran and Kawsar Genetic Research Center, 40 were suspected of having a large deletion in β-globin gene cluster. The including criteria were hematological findings such as low blood indices (MCV <80 fl and MCH <27 pg), normal HbA2 and raised or normal HbF. Genomic DNA was extracted from peripheral blood. A Real-time PCR assay was developed using comparative threshold cycle (Ct) method for analysis of gene copy number. In addition, gene dosage was analyzed using MLPA method.

 

 Results

 Real-time PCR results for quantitative analysis of Beta, Delta, G-gamma genes showed the ratio (2-ΔΔCt) of 0.96 ± 0.18 for normal individuals and 0.58 ± 0.04 for carriers of deletions in beta globin gene cluster. MLPA results showed nearly 50% reduction in the height of the peaks corresponding to regions of deletions.

 

 Conclusions

 MLPA results confirmed the presence of the same deletions detected by Real-time PCR in all of the carrier individuals. It would be ideal to combine these quantitative assays to confirm corresponding results for accurate diagnosis of known and unknown deletions in beta thalassemia carriers.

  

 Key words : beta-Thalassemia, beta-Globins, Gene Deletion

 


N. Kh. Karimi, L. Kokabee, Z. Badiee, M. Shahabi, S.a. Banihashem, S. Zeinali, K. Parivar, M. Karimipoor,
Volume 7, Issue 1 (Spring 2010)
Abstract

 

  Abstract

 Background and Objectives

 Hemophilia B, Christmas disease, is an X-linked recessive bleeding disorder caused by the functional deficiency of blood coagulation factor IX (FIX). The disease is caused by heterogeneous mutations in the factor IX gene (factor IX). FIX is a vitamin K-dependent glycoprotein that plays a key role in the coagulation cascade. The aim of this study was to make molecular analysis of factor IX gene and evaluate genotype-phenotype correlation in 14 hemophilia B patients from Khorasan Razavi province.

 

 Materials and Methods

 Fourteen unrelated hemophilia B patients were included in the study. The patients had high PTT, normal PT, low factor IX activity (less than 30%), and normal FVIII activity. After obtaining informed consent, genomic DNA was extracted from the peripheral blood leukocytes by standard methods. Polymerase chain reaction (PCR) and single strand conformation polymorphism (SSCP) were performed to obtain a full scan of all functionally important regions of the FIX gene. DNA sequencing was done on samples with a definite band shift in SSCP. In addition, Haplotypes were constructed using four markers (DdeI, TaqI, MnlI and HhaI).

 

 Results

 The sequencing results showed 9 missenes mutations, 3 stop codons, and 1 insertion. Two of the mutations not having been reported in the database so far were novel. Malmo polymorphism (Ala148Thr) was found in two patients.

 

 Conclusions

 This study confirms the heterogeneity of factor IX gene mutations in northeast hemophilia B patients. The obtained data could be used both in tracking female carriers in families and in making decisions about prenatal diagnosis.

 

 Key words : Hemophilia B, SSCP, Mutation, Factor IX

 


Dr. B. Zarbakhsh, E. Farshadi, A. Ariani Kashani, Dr. M. Karimipoor, Dr. A. Azarkeivan, R. Habibi Pourfatideh, Dr. M.s. Fallah, Dr. F. Maryami, A.r. Kordafshari, Z. Kaeini Moghadam, Dr. H. Bagherian, Dr. S. Zeinali,
Volume 7, Issue 2 (Summer 2010)
Abstract

  Abstract

  Background and Objectives

  There is a large number of couples who are considered potential carriers of alpha or beta-thalassemia. The exact determination of gene defect for thalassemia carriers is essential for premarital screening genetic counseling. In this study, we conducted a molecular study of those suspected of carrying alpha-thalassemia mutated genes in order to detect potential deletional and non-deletional mutations in the alpha globin gene cluster.

 

  Materials and Methods

  In this study, those suspected of having mutation in alpha-globin gene cluster with MCV < 80 fl, MCH < 27 pg, normal serum iron, and HbA2 were selected from those referred to Pasteur Institute of Iran. Four common deletional mutations and non-deletional mutations were studied using multiplex gap-PCR, ARMS-PCR, and direct sequencing.

 

  Results

  One hundred and forty samples with above criteria entered the study with 126 (90%) cases showing at least one mutation. Study of 4 common deletional mutations using multiplex gap-PCR revealed at least one deletion in 99 (70.71%) cases. Non-deletional mutations were found using ARMS-PCR or direct sequencing in 27 (19.28%) cases. Nine different mutations were found in the samples with –α3.7 being the most common deletion in 100 (35.71%) alleles out of 280 studied chromosomes followed by -α5nt in 25 (8.93%) alleles as the next most common.

 

  Conclusions

  We used direct sequencing to characterize more suspected carriers of alpha thalassemia. However, using other methods like real-time PCR and multiplex ligation-dependent probe amplification (MLPA) for gene dosage study of alpha-globin gene cluster could help find other non-common deletions.

 

  Key words :

  Abstract

 Background and Objectives

 There is a large number of couples who are considered potential carriers of alpha or beta-thalassemia. The exact determination of gene defect for thalassemia carriers is essential for premarital screening genetic counseling. In this study, we conducted a molecular study of those suspected of carrying alpha-thalassemia mutated genes in order to detect potential deletional and non-deletional mutations in the alpha globin gene cluster.

 

 Materials and Methods

 In this study, those suspected of having mutation in alpha-globin gene cluster with MCV < 80 fl, MCH < 27 pg, normal serum iron, and HbA2 were selected from those referred to Pasteur Institute of Iran. Four common deletional mutations and non-deletional mutations were studied using multiplex gap-PCR, ARMS-PCR, and direct sequencing.

 

 Results

 One hundred and forty samples with above criteria entered the study with 126 (90%) cases showing at least one mutation. Study of 4 common deletional mutations using multiplex gap-PCR revealed at least one deletion in 99 (70.71%) cases. Non-deletional mutations were found using ARMS-PCR or direct sequencing in 27 (19.28%) cases. Nine different mutations were found in the samples with –α3.7 being the most common deletion in 100 (35.71%) alleles out of 280 studied chromosomes followed by -α5nt in 25 (8.93%) alleles as the next most common.

 

 Conclusions

 We used direct sequencing to characterize more suspected carriers of alpha thalassemia. However, using other methods like real-time PCR and multiplex ligation-dependent probe amplification (MLPA) for gene dosage study of alpha-globin gene cluster could help find other non-common deletions.

  

 Key words : alpha-Thalassemia, mutation, PCR, Iran

 


A. Rajabi, A. Arab, Dr. M. Karimipoor, Dr. S. Kaviani, Dr. Kh. Arjmandi, Dr. S. Zeinali,
Volume 8, Issue 1 (Spring 2011)
Abstract

  Abstract

 Background and Objectives

 The molecular basis for thalassemia intermedia (TI) is determined in most thalasemia affected countries, but in Iran there has been no perfect investigation so far. This report is the results of the first step of a comprehensive analysis on molecular basis of TI in Iran. Two most important factors influencing the phenotype of TI, i.e. beta globin gene mutations, G γ XmnI polymorphism, and genotype/phenotype correlation in these patients were analysed.

 

 Materials and Methods

 In an experimental pilot study, 42 TI patients who referred to Ali-Asghar Hospital were selected and genomic DNA was extracted by salting out method. The ARMS-PCR technique was performed to detect the most prevalent β thalassemia mutations in Iran: IVSII-1 (G>A). Direct DNA sequencing was performed on the samples which had at least one unidentified allele. Simultaneously, G γ XmnI polymorphism was determined using PCR-RFLP procedure. Afterwards, the association of this polymorphism with TI was analyzed.

 

 Results

 Among 76 chromosomes, IVSII-1 was the most frequent mutation detected with 42 alleles (55.26%). Totally, 66 β globin alleles (86.84%) were β0 and 7 (9.21%) β+. In addition, 61 chromosomes (80.26%) were positive for XmnI polymorphism. This polymorphism was in strong linkage to β0 mutations, mainly IVSII-1. No cases with IVSII-1 mutation were XmnI-/-.

 

 Conclusions

 It seems that the presence of XmnI polymorphism may play an important role in reducing the clinical severity of thalassemia in patients with severe β0 alleles. However, other genetic factors should be also investigated.

 


F. Hashemi-Gorji, Dr. M. Hamid, A. Arab, A. Amirian, Dr. S. Zeinali, Dr. M. Karimipoor,
Volume 8, Issue 3 (Autumn 2011)
Abstract

  Abstract

 Background and Objectives

 High fetal hemoglobin (HbF) levels have a major impact on the hemoglobin disorders, i.e. β-Thalassemia. Increased HbF production ameliorates the disease severity. Three loci—HBS1L-MYB intergenic region on chromosome 6q23, BCL11A on chromosome 2p16, and the γ-globin gene on chromosome 11 account for up to 50% of the variations in HbF levels in patients with sickle cell anemia, thalassemia and healthy adults. In the present study, we evaluated the relationship between some polymorphisms on HBS1L-MYB BCL11A loci and increased HbF levels in thalassemia patients and normal subjects.

 Materials and Methods

 In this case-control study, three common polymorphisms among 50 β-thalassemia patients with increased HbF and 47 healthy individuals with normal HbF by using PCR-RFLP were genotyped: rs4895441, rs11886868, and rs28384513. Enzymatic digestion was performed by RsaI, MboII, and BstXI, respectively. Correlations with high levels of HbF were performed with a Chi-square test by using SPSS 16 and SNP analyzer2.

 Results

 Mutant allelic frequencies were 0.245, 0.521 and 0.309 in healthy and 0.3, 0.52 and 0.28 in patient for rs4895441, rs11886868 and rs28384513, respectively. Significant relationship was not observed among three polymorphisms studied in healthy volunteers and β-Thalassemia major patients with increased HbF levels and P-value allelic and genotypic was higher than 0.05 at three SNPs.

 Conclusions

 In spite of previous reports, evaluation of polymorphisms at the BCL11A and HBS1L-MYB loci in this study did not show up a significant correlation with increased HbF. Other polymorphisms might have a role in increasing HbF in our population.


Dr. B. Zarbakhsh, F. Eghbalpour, E. Farshadi, Dr. Ms. Fallah, Dr. M. Karimipoor, Z. Kaeini Moghadam, Dr. S. Zeinali,
Volume 9, Issue 4 (Winter 2013)
Abstract

 

  Abstract

 Background and Objectives

 Based on the previous researches performed in Iran and worldwide, there is a significant number of subjects carrying alpha-thalassemia gene defect. Therefore, we aimed to assess its frequency amongst a random population selected from premarriage couples who entered the national prevention program of β -thalassemia major at Tehran.

 

 Materials and Methods

 Blood samples were collected from 625 randomly selected individuals following obtaining written informed consent through eight cooperating primary health care centers (PHC) in Tehran. Primarily, CBC testing was performed. Subsequently, blood samples underwent DNA extraction and gene-specific amplification using ARMS PCR and Multiplex Gap PCR followed by DNA sequencing of negative samples to identify potentially missed alpha-globin gene mutations.

 

 Results

 According to CBC results, 147 individuals were considered normal 479 remaining blood samples underwent molecular assessment which revealed that 95 samples possessed a minimum of one deletional mutation in the alpha-globin genes, resulting in a frequency of 19.83% in the tested population (CI 95% 11.81-27.85). Furthermore, 50 samples with normal hematological indices (MCH>27, MCV>80) were tested for triplication mutation and revealed to be negative.

 

 Conclusions

 It has previously been established that Multiplex Gap PCR is capable of detecting 65% of alpha globin gene defects in blood samples with abnormally low hematological indices (MCH<27, MCV<80). Hence, using bio-informatic techniques with generalization of our results to the entire population of Tehran showed the true frequency of alpha-globin gene defect to be actually 29.90% (CI 95% 22.40-37.40) at the population level .

 



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