[Mutations in genes affecting fertility of men - current routine laboratory genetic diagnostics and searching for more DNA segments and genes influencing spermatogenesis]. / Mutace v genech ovlivnujících plodnost muzu - soucasná rutinní laboratorní genetická diagnostika a hledání dalsích úseku DNA a genu, ovlivnujících spermatogenezi.

OBJECTIVE: To present the results of molecular genetics analysis in men with reproductive disorders focusing on the DNA segments and genes which affect spermatogenesis. DESIGN: Original article. SETTING: Institute of Biology and Medical Genetics of the First Faculty of Medicine and General Teaching Hospital, Prague. METHODS: One hundred and twenty-three patients identified with a fertility disorder were screened for mutations of the CFTR gene. In all patients were performed cytogenic analysis and assessment of Y-chromosome microdeletions. In 107 patients where the fertility was not detected by routine examination we performed an analysis for X-chromosome microdeletions (CNV64, CNV67, CNV69) and in certain genes necessary for normal spermatogenesis (AGFG1, CAPZA3, CNTROB, HOOK1, GOPC, SPATA16). RESULTS: Our results did not reveal any negative efffects of X-chromosome microdeletion on spermatogenesis. Analysis of six genes showed in two patients in gene SPATA16 a homozygotic haplotype [1526C>T + 1577T>C] which can be most probably responsible for the fertility in two examined patients. CONCLUSION: According to our results we do not recommend introduction of X-chromosome microdeletions assays in areas CNV64 , CNV67 and CNV69 into routine diagnostic. Regarding the selected genes affecting spermatogenesis, our results showed that homozygotic haplotype [ 1526C>T + 1577T>C] in SPATA16 gene is very likely responsible for infertility in two of our patients. The above mentioned haplotype deserves attention in the investigation of male infertility.

[Dilemma of the results interpretation of molecular genetic analysis with a focus on CFTR gene mutations in men with reproductive disorders and in gamete donors]. / Problematika interpretace výsledku molekulárne genetických vysetrení se zamerením na mutace v CFTR genu u muzu s poruchami reprodukce a u dáru/dárkyn gamet.

OBJECTIVE: The presentation of the results of molecular genetics analysis in men with reproductive disorders and in gamete donors with a focus on interpretation of the results CFTR gene analysis. DESIGN: Original article. SETTING: Institute of Biology and Medical Genetics of the First Faculty of Medicine and General Teaching Hospital. METHODS: We examined 164 men with reproductive disorders for 36 selected mutations in CFTR gene including T(n) polymorphism and for Y chromosome microdeletions. As well we examined mutations in CFTR gene including T(n) polymorphism in 104 gamete donors. RESULTS: We detected microdeletions in AZF region in 3 cases of affected men and in other 3 casses we found mutation F508del (heterozygotes) in CFTR gene with T5 variant in trans position. Except this we detected in 5 affected men "only" heterozygous mutations in CFTR gene and in 12 men "only" the T5 variant in heterozygous level. Among gamete donors we found 3 heterozygotes for mutation F508del and 11 heterozygotes for T5 variant. CONCLUSION: In infertile men and in gamete donors we recommend to examine not only the "classical" mutations in CFTR gene but also the relatively frequent T5 variant, which can be in certain conditions considered as a pathogenic mutation. It's necessary to rule out the carriers of T5 variation from gamete donors.

Patterns of deletions and the distribution of breakpoints in the dystrophin gene in Czech patients with Duchenne and Becker muscular dystrophy (statistical comparison with results from several other countries).

Deletion pattern analysis of the dystrophin gene was performed in 115 unrelated Czech patients with Duchenne and Becker muscular dystrophy. In 50 patients (43.5% of the analysed patients) exon deletions were detected by routinely performed multiplex PCR for 18 selected exons and for the area of musclespecific promoter of the dystrophin gene. All startpoints and endpoints of deletions (100 breakpoints) were detected using PCRs for another 29 exon areas of the dystrophin gene (altogether primers for 47 different exons were used). Most of the breakpoints were found in introns 44 (16% of breakpoints), 47 (14%) and 50 (8%). The comparison of distributions of breakpoints in the area of the main hot spot of the dystrophin gene (introns 43-52) was made (chi 2 test in a contingency table) in six different populations from the Czech Republic, Bulgaria, Hungary, Italy, Turkey and India. In compared populations, statistically significant differences were found by the pooled test. No significant difference between the Czech population and other studied populations was found by pair comparisons. On the other hand, pair comparisons revealed significant differences between populations from Bulgaria and Hungary, Bulgaria and Turkey, Hungary and Italy. The results of the presented study support the theory suggested by other authors that specific differences in intron sequences of the dystrophin gene can exist between different populations, possibly as a result of a genetic drift.