The Study On Application Of Detection Of Y Chromosomal Microdeletion In Patients With Idiopathic Azoospermia Or Oligospermia

Objective To found the molecular detection methods of the Y chromsome microdeletions in patients with idiopathic oligozoospermia and azoospermia. By using the methods to confirm the microdeletion rate of these patients. Studied the relations between azoospermia factor (AZF) with the patients and the distribution rule of microdeletions of AZF in these patients and to evaluate on the patients who can be applied ICSI and wether they must process prenatal diagnosis after ICSI.Methods Acording to EAA/EMQN best practice guidelines for molecular diagnosis of Y-chromosomal microdeletions(State of the art 2004) and refrence ,we select eight sequence-tag-sites in Y chromosome including AZFa,b,c and d regions,six of which to be detected with mutiplex PCR,two of which to be detected with single PCR.51 idiopathic oligozoospermia and azoospermia patients and 10 normal fertility men were detected.Results 1.In the peripheral blood samples of the 51 infertile and 10 fertile men,the primer of SRY gene was used as positive controls for PCR.The express of SRY gene was evaluated in order to have an internal control for the DNA isolation and efficiency of the PCR method.2. Four STS markers,including the internal control the SRY gene, can be distinguished and have 72-346bp ladder.3.Amplified products of SRY,AZFa,AZFb and AZFc were expressed under each photograph.The microdeletion of AZF region was not found in 10 normal subjects with offspings.4.Among 51 patients,there were AZFb microdeletions: sY134 microdeletion to be detected in patient 08, sY127microdeletion to be detected in patient 34.All of the ratio of Y chromosomal microdeletions was 3.92%(2/51),the ratio of microdeletion in azoospermia patients was 8%(2/25),the ratio of microdeletion in oligospermia patients was 0(0/26).5.To confirm our results,all of the 2 samples were amplified three times. The microdeletions of sY127,sY134 were approved if the PCR product of SRY gene was of the expected size and sY127 or sY134 were negative only after three times amplification.6.There were no AZFd microdeletions in the 51 idiopathic oligozoospermia and azoospermia patients.Conclusions 1.Y microdeletions have relation with idiopathic oligospermia and azoospermia patients.The ratio of Y microdeletions in patients with idiopathic oligozoospermia and azoospermia was 3.92%.2.The distributions of Y chromosome microdeletions have relations with idiopathic oligozoospermia and azoospermia patients.In our study,two of the AZFb microdeletions were idiopathic azoospermia patients.3. AZF deletions could be transmitted by vertical transmission,and would probably result in infertile sons. In cases of idiopathic oligozoospermia and azoospermia, detection of correlation gene may play an important role in the increase of prepotency ratio before starting artificial insemination with husband's semen and Intracytoplasmic sperm injection.4.We have to study further the type and the frequency of AZFd microdeletions, and the relations between AZFd microdeletions with the patients of idiopathic oligozoospermia and azoospermia .