Study On The Genetic Diagnosis Of Two Typical Teratozoospermia

Objective: Teratozoospermia refers to male infertility in which the percentage of normal sperm morphology is lower than the reference value(2010,the WHO specified value was 4%).Autosomal recessive inheritance is the main cause of teratozoospermia.Have been reported of sperm abnormalities including sperm flagella Multiple morphological variations,Large headed multiflagellar spermatozoa,Globozoospermia and Acephalic spermatozoa syndrome.In the first part,the classification and clinical manifestations,pathogenesis and genetic etiology,diagnosis and differential diagnosis,genetic testing and genetic counseling and assisted reproductive outcomes of teratospermia were systematically and comprehensively described,which laid a good foundation for the study of teratospermia.In the second part,the sperm morphology of 1532 patients was analyzed,so as to provide important reference basis for clinical diagnosis.In the third part,25 patients with MMAF were studied to determine the cause of their disease from the perspective of genetics,so as to lay a foundation for clinical treatment and genetic counseling of patients.In the fourth part,20 cases of globozoospermia were studied to clarify the pathogenesis from the aspect of genetics,so as to lay a foundation for the clinical treatment and genetic counseling of the patients.Methods:In the first part,literature on teratospermia was collected and reviewed from the NCBI database to summarize the genetic causes,diagnosis and treatment,and assisted reproductive outcomes of teratospermia.In the second part,1532 patients came to our hospital for at least three times of semen routine examination and sperm morphology analysis.The third part,in view of the 25 patients with MMAF,collection the peripheral blood of patients.(1)Based on multiple PCR targeted sequencing technology to MMAF four pathogenic gene(DNAH1 / CFAP43 / CFAP44 / CFAP69)all exons location point mutation detection.(2)Test the entire exon region of MMAF patients through whole-eome sequencing(WES).(3)By Polymerase Chain Reaction amplification,Sanger sequencing and bioinformatics analysis software validation mutation loci.In the fourth part,peripheral blood was collected from 20 patients with round head spermia.(1)PCR amplification was used to detect the variation site of DPY19L2 total exon region.(2)WES was used to detect the variation of the whole exon region in patients with round head spermia.(3)The mutation sites were verified by Sanger sequencing and bioinformatics analysis software.Results: Firstly,the sperm morphology of 1532 patients was studied,and 600 patients were finally diagnosed as male infertility patients,of which 60 were diagnosed as teratozoospermia.Unfortunately,15 of the 60 patients with teratozoospermia were unable to obtain peripheral blood,so only 45 patients with teratospermia(25 patients with MMAF and 20 patients with globozoospermia)who obtained peripheral blood were examined for genetic diagnosis in this study.Secondly,25 MMAF patients were studied for gene diagnosis.Among the four unrelated patients,one new homozygous shift code mutation(c.11023delT)of DNAH1 gene and three new complex heterozygous mutations were found.Finally,genetic diagnosis of 20 patients with globozoospermia was studied,and homozygous deletion of DPY19L2 gene and new homozygous missense mutation(c.1889G>A)were found in 5unrelated patients.Conclusion:In this study,it was found that sperm morphology is an essential and important index in the diagnosis of male infertility,which provides an important reference for doctors to make diagnosis.On the other hand,the genetic etiology of 4patients with MMAF and 5 patients with globozoospermia was identified.DNAH1 and DPY19L2 gene mutations not only enrich the gene mutation spectrum and facilitate the genotype-phenotype correspondence,but also provide an important basis for the subsequent clinical evaluation,assisted reproduction and genetic counseling of patients.