Scanning of β‐globin gene for identification of β‐thalassemia mutation in Romanian population

Abstract

Abstractβ-Thalassemia is uncommon (0.5%) in the Romanian population, but it must be considered in the differential diagnosis of hypochromic anemia. The molecular characterization of β-thalassemia is absolutely necessary for molecular diagnosis, as well as any genetic epidemiological study in this region. Molecular analyses consist of mutation detection by molecular scanning of β-globin gene. This gene has 3 exons and 2 introns, involved in β-thalassemic pathogenesis. Clinical application of DNA analysis on β-thalassemic chromosomes allowed characterization of 29 persons with different β-thalassemia mutations among 58 patients with anemia. The experimental strategy was based on sequential PCR amplification of most of the β-globin gene and running on denaturing gradient gel electrophoresis of amplification products. Definitive characterization of mutations in samples identified with shifted DGGE patterns was performed ARMS-PCR and/or PCR-restriction enzyme analysis methods. Eight different β-thalassemia alleles were identified, the most common being IVS I-110 (G-A) and cd 39 (C-T). Comparison of overall frequency of mutations in the neighboring countries, shows that these results are in the frame of overall distribution of these mutations in Mediterranean area, especially in Greece and in Bulgaria. Molecular diagnosis is useful for differentiating mild from severe alleles, for genetic counseling, as well as for mutation definition in carriers, identified by hematological analysis necessary for prenatal testing and genetic counseling.

Thalassemia major phenotypes secondary to the association of β 5′UTR +20(C→T) allele with β 39(C→T)

“…Interestingly, the +20 (C?T) b + mutation was not associated with IVSII-745(C? G) in cis, as it has been described in previously reported cases (4)(5)(6).…”

HIGH SENSITIVE PCR METHOD FOR DETECTION OF PATHOGENIC Leptospira spp. IN PARAFFIN-EMBEDDED TISSUES

“…Fifty seven paraffin-embedded organ samples (liver, kidney, lung, brain, heart or spleen) from 24 dead patients with suspicion of leptospirosis, received at the National Reference Laboratory of Leptospires (IPK) during 2012, were tested firstly by a PCR method detecting sample inhibition and this amplified a 268 bp of the gene encoding human β-globin 15 ; after, the non-inhibited samples were subjected to the application of both PCR methods, LipL32 PCR and 23S PCR.…”

Molecular study and genotype/phenotype correlation of β thalassemia in Malaysia

“…, 2004; Tan, Tan & Thong, 1999). If ARMS‐PCR failed to detect any mutation, the sample was analyzed by direct sequencing as previously described (Talmaci et al. , 2004).…”

“…The b-globin genes were analyzed using the ARMS-PCR method as previously described (Weatherall & Clegg, 2001), tailored to detect the seven b thalassemia mutations common in Malaysia, namely CD41/42, CD71/72, IVS1-5, IVS1-1, CD26, IVS2-654, and CD17 (Tan et al, 2004;Tan, Tan & Thong, 1999). If ARMS-PCR failed to detect any mutation, the sample was analyzed by direct sequencing as previously described (Talmaci et al, 2004).…”