Rapid screening of multiple beta-globin gene mutations by real-time PCR on the LightCycler: Application to carrier screenig and prenatal diagnosis of thalassemia syndromes

Background: Hemoglobinopathies are priority genetic diseases for prevention programs. Rapid genotype characterization is fundamental in the diagnostic laboratory, especially when offering prenatal diagnosis for carrier couples. Methods: As a model, we designed a protocol based on the LightCycler(TM) technology to screen for a spectrum of beta-globin gene mutations in the Greek population. Design was facilitated by dual fluorochrome detection and close proximity of many mutations. Three probe sets were capable. of screening 95% of beta-globin gene mutations in the Greek population, including IVSII-7 745C–>G, HbS, Cd5-CT, Cd6-A, Cd8-AA, IVSI-1G–>A, IVSI-5G–>A, IVSI-6T–>C, IVSI-110G–>A, and Cd39 C–>T. Results: The protocol, standardized by analysis of 100 beta-thalassemia heterozygotes with known mutations, was 100% reliable in distinguishing wild-type from mutant alleles. Subsequent screening of 100 Greek beta-thalassemia heterozygotes with unknown mutations found 96 of 100 samples heterozygous for 1 of the 10 mutations, although melting curves were indistinguishable for mutations HbS/Cd6 and IVSI-5/IVSI-1, indicating a need of alternative methods, for definitive diagnosis. One sample demonstrating a unique melting curve was characterized by sequencing as Cd8/9+G. Three samples carried mutations outside the gene-region covered by the probes. The protocol was 100% accurate in 25 prenatal diagnosis samples, with 14 different genotype combinations diagnosed. The protocol was also flexible, detecting five beta-globin gene mutations from other population. groups (IVSI-1G–>T, IVSI-5G–>C, IVSI-116T–>G, Cd37 TGG–>TGA, and Cd41/42 - TCTT). Conclusions: The described LightCycler system protocol can rapidly screen for many beta-globin gene mutations. It is appropriate for use in many populations for directing definitive mutation diagnosis and is suited for rapid prenatal diagnosis with low cost per assay. (C) 2003 Arnerican Association for Clinical Chemistry

Association of repeat polymorphisms in the estrogen receptors alpha, beta (ESR1, ESR2) and androgen receptor (AR) genes with the occurrence of breast cancer

Genetic variation in genes involved in estrogen biosynthesis, metabolism and signal transduction have been suggested to play a role in breast cancer. To determine the possible contribution of genetic variation in the ESR1 (ER-α), ESR2 (ER-β) and AR genes in breast cancer risk the -1174(TA)7-27, c. 1092+3607(CA)10-26 and c. 172(CAG)6-40 repeat variants were studied in a case-control study of 79 women with sporadic breast cancer and 155 controls. No significant difference was observed in the frequency distribution of -1174(TA)7-27 in the ESR1 gene between patients and controls, while a significant difference was observed for repeat polymorphisms c. 1092+3607(CA)10-26 in the ESR2 gene and c. 172(CAG)6-40 in the AR gene (p≤0.0001). A significantly decreased odds ratio (OR) for breast cancer risk was observed in individuals having the LL and the SL genotypes for both the ESR2 (OR=0.010, 95% CI 0.003-0.036, p<0.001; OR=0.013, 95% CI 0.004-0.040, p<0.0001, respectively) and the AR gene (OR=0.040, 95% CI 0.011-0.138, p<0.0001; OR=0.189, 95% CI 0.10-0.359, p<0.0001, respectively), compared to SS genotype. The protective effect of these genotypes remained evident even after adjustment for various risk factors (BMI, age, age at menarche and menopause, family history). In conclusion, an association for breast cancer risk between short (SS) alleles for the repeat variants of the ESR2 and AR genes was found in women of Greek descent. © 2007 Elsevier Ltd. All rights reserved

Environmental Assessment of Organic Waste and Domestic Wastewater Management in Decentralised Communities

Environmental Assessment of Organic Waste and Domestic Wastewater Management in Decentralised Communitie

Analysis of PRNP gene codon 129 polymorphism in the Greek population

Creutzfeldt-Jakob disease (CJD) is a fatal transmissible neurodegenerative prion disease with a rapid progression comprising familial, sporadic, iatrogenic and variant forms. A polymorphism at codon 129 of PRNP gene has been implicated in the development of variant CJD. We examined Met/Val allele frequencies and the genotype distribution, with respect to the polymorphic codon 129 of PRNP gene in 348 healthy individuals from the region of Athens, Greece. The following genotype frequencies were observed in the Greek population: Met/Met 50%, Met/Val 39% and Val/Val 11%. The presence of the Methionine allele frequencies in various European populations, according to the published data, increases gradually from northwestern to southeastern countries, implying the presence of a cline. The distribution of genotypes of Met homozygotes displays random declination across the 10 compared populations. The observed higher frequency of Met homozygotes at codon 129 does not necessarily suggest that these populations are at increased risk of developing CJD

Editorial - Sustainable waste and wastewater management

Μη διαθέσιμη περίληψηNot available summarizationPresented on

Editorial - Waste management

Μη διαθέσιμη περίληψηNot available summarizationPresented on: Journal of Environmental Managemen