Gene Mutation Of FSHR And LHCGR In The Patients With Hypergonadotrophic Amenorrhea Or Poor Ovarian Reaction And Mutation Screening And Prenatal Diagnosis In 14 Families With Tuberous Sclerosis Complex

Objective to identify the mutations of follicle stimulating hormone receptor and luteinizing hormone receptor in patients who was diagnosised as hypergonadotrophic amenorrhea or poor ovarian reaction in COH and patients whose FSH are unexplained beyond the normal level. Then to apply the gene detection results to their reproduction.Methods We collected the peripheral blood from 16 persons and obtain their gDNA. PCR and DHPLC were applied to detect the mutations in most parts of FSHR and LHCGR gene. DNA sequencing was applied to affirm the mutation detected by DHPLC. And some exons such as FSHRe5, FSHRe10-3, FSHRe10-5 and LHCGRe1, LHCGRe9 were detected with PCR and direct sequencing because of hard curve with DHPLC.Results two FSHR mutations were found in 16 patients with the PCR-DHPLC-sequencing and PCR-sequencing. Patient one had a homozygous mutation (C1319A) which changes the Alanine to Glutamate at CDS440. And it is a novel mutations unrecorded by HGMD. The PCR-restriction enzyme digestion showed that a novel HpyAV enzyme digestion location appeared in patient 1. However there was no change in HpyAV enzyme digestion location of the DNA samples obtained from 100 unrelated normal Chinese women with fertility proved. Patient two had a deletion mutation (c.419delA) which generates a premature stop codon at CDS 155. No LHCGR mutation was detected and other 14 persons had not any mutation found.Conclusion Two FSHR mutations found in two women may be the reason why their follicular development was blocked and their FSH/LH levels were unexplained increased. C1319A and c.419delA both are novel mutations unrecorded by HGMD. And c.419delA is the first FSHR deletion mutation ever found. Our found are the first FSHR mutations found in Chinese people. And we also make clear that which patient should conduct FSHR and LHCGR gene mutation detection.Objective to screen for mutations in TSC patients and perform prenatal diagnosis to prevent the birth of children with TSCMethods PCR-DHPLC-sequencing was used to screen TSC1 and TSC2 mutations in 16 patients of 14 pedigrees visited our hospital in last five years. T/A cloning was performed when necessary. For novel mutations, one hundred unrelated normal individuals were screened to exclude the possibility of being polymorphism.Results Eleven mutations were found in 13 patients out of 16 patients from 14 pedigrees with nine being novel mutations unrecorded by HGMD. The mutations included 4 deletion mutations (TSC2 c.1143delG, TSC2 c.1957-1958delAG, TSC2 c.4541-4544 delCAAA and TSC1 c.2672delA),2 splicing mutations (TSC2 Intron27+1 G> A, and TSC2 Intron5+1 G>A),3 missense mutations (TSC2 R1743Q, TSC2 L847P and TSC1 N762S) and 2 insertion mutations (TSC2 c.4918insCGCC and TSC2 c.910insCT). Only 2 TSC1 gene mutations (N762S and c.2672delA) were detected, and the other 9 were all TSC2 gene mutations. Prenatal diagnosis for 6 pedigrees (1,2,3,4,8 and 9) indicated that the fetus of family 1,2,3,4 or 8 did not carry the mutation existed in his or her family, and the fetus of family 9 carried the same mutation as his or her mother.Conclusion The mutation rate of tuberous sclerosis was 81.25% (13/16) among patients in our hospital. The ratio of TSC2 mutations to TSC1 ones was about 1:1 in the familial cases and TSC2 mutations were more common in sporadic cases. Nine mutations identified in this study were novel, which indicates high frequency of de novo mutation associated with TSC. Our data demonstrated that birth of TSC children for those with familial history of TSC could be prevented via prenatal diagnosis.