Molecular characterization of hereditary hemochromatosis in Matera province

Hereditary hemochromatosis is a common recessive disorder characterized by ironoverload that leads to multi-organ failure. The classic disease is associated with HFE gene mutation so chromosome6; other forms were related to genes involved in iron transport, storage, and regulation. The aim of this work was directed to investigate: a) thefrequency of the main mutations; b) the association between mutations and case so hemochromatosis in Matera province (Basilicata, Italy).The experimental design provided DNA extraction from whole blood of 195 individuals resident in Matera district. DNA amplification by multiplex PCR, reverse dot-blot on nitrocellulose strips, statistical data analysis were performed to analyze the presence of 11 HFE gene mutations ( V53M, V59M, H63D, H63H, S65C, Q127H, E168X, E168Q, W169X, C282Y, Q283P), 4Transferrin Receptor mutations (Y250X, E60X, M172K, the AVAQ594-597) and 2 Ferroportin mutations (N144H, V162). 117subjects(60%)of the analyzed population were healthy, 62 (32%) were heterozygous (57 cases H63D/wt,4 C282Y/wt, 1S65C/wt). There were 16 patients with hemochromatosis (10cases H63D/H63D, 2C282Y/C282Y, 4H63D/C282Y). Genotypic frequencies showed important differences in comparison with data on Italian population, where C28Y homozygous are more frequent and they manifest more severe iron over load than the other HFE genotypes

Molecular Methods for a Correct Diagnosis of Multiple HPV Infections and Clinical Implications for Vaccine.

Introduction: The human papillomavirus (HPV) family is characterized by minimal geno- typic differences corresponding to different virus types. The aim of this study was to detect the HPV coinfections and the inner genotype in a series of 336 cervical-vaginal samples. Methods: A total of 336 cervical-vaginal samples were taken from 2007 to 2009 using specific molecular techniques such as molecular sequencing and hybridizations. The ge- nome amplification of the L1 open reading frame was analyzed by real-time polymerase chain reaction; direct sequencing was performed by SYBR green fluorescent molecule and degenerate primers MY09 and MY11. The HPV genotyping was accomplished via oli- gonucleotide probe hybridization. The phylogenetic correlations in coinfections were an- alyzed by sequence homology of the L1 genomic region. Identified genotypes were then compared. Results: Human papillomavirus positivity was observed in 125 cases (37.2%), with 21 cases (16.8%) of HPV presence in coinfections. Coinfections involved HPV 16 geno- type (8 cases) and HPV 18 (5 cases). The HPV 16 infection was mainly associated with genotypes with a lower-than-broad sequence homology, so the HPV 18 was linked to genotypes represented in the opposite phylogenetic tree. Conclusions: The combined and steady use of diagnostic procedures, such as real-time polymerase chain reaction, molecular hybridization, direct sequencing, and HPV geno- typing test, allow accurate diagnosis of monoinfections and coinfections. This may faciliate the development of specific viral tests and prophylactic anti-HPV vaccines

Analysis of cystic fibrosis gene mutations in children with cystic fibrosis and in 964 infertile couples within the region of Basilicata, Italy: a research study

Introduction: Cystic fibrosis is the most common autosomal recessive genetic disease in the Caucasian population. Extending knowledge about the molecular pathology on the one hand allows better delineation of the mutations in the CFTR gene and the other to dramatically increase the predictive power of molecular testing. Methods: This study reports the results of a molecular screening of cystic fibrosis using DNA samples of patients enrolled from January 2009 to December 2013. Patients were referred to our laboratory for cystic fibrosis screening for infertile couples. In addition, we identified the gene mutations present in 76 patients affected by cystic fibrosis in the pediatric population of Basilicata. Results: In the 964 infertile couples examined, 132 subjects (69 women and 63 men) resulted heterozygous for one of the CFTR mutations, with a recurrence of carriers of 6.85%. The recurrence of carriers in infertile couples is significantly higher from the hypothetical value of the general population (4%). Conclusions: This study shows that in the Basilicata region of Italy the CFTR phenotype is caused by a small number of mutations. Our aim is to develop a kit able to detect not less than 96% of CTFR gene mutations so that the relative risk for screened couples is superimposable with respect to the general population. Keywords: Cystic fibrosis, Cystic fibrosis conductance transmembrane regulator, Screening in infertile couples, Mutation analysis, Polymerase Chain Reactio