Comparison between Real Time PCR and Gel based PCR Technique in Diagnosis Y Chromosome microdeletions in infertile azoospermia males in the Iraqi Kurdish Population (original) (raw)
Related papers
Study of Y-Chromosome Microdeletions in Azoospermic Infertile Males using Multiplex PCR Analysis
Biosciences, Biotechnology Research Asia, 2018
The infertility affects about 15% of couples and male factors being responsible about 40-50%. In male infertility, genetic abnormalities of Y chromosome play crucial role in spermatogenesis defect. Y chromosome q arm having Azoospermia factor region (AZFa, AZFb, and AZFc) are most important for spermatogenesis. Here, we investigated the frequencies of Y-chromosome microdeletions using three sets of multiplex PCR in idiopathic cases of azoospermia. We studied a total of 110 infertile male with non-obstructive azoospermia subjects & 50 fertile control subjects. All DNA samples were used for Y chromosome microdeletions analysis by using 11 STS markers in three different multiplex PCR of AZF regions. Out of 110 infertile azoospermic males, 14 (12.72%) infertile males showed partial deletion of AZF regions using three sets of multiplex PCR group. In the AZF microdeletions of infertile males, individually AZFc region was the most deletions sites (10%) followed by AZFb (6.36%) and AZFa (1....
Prevalence of Y chromosome microdeletion in azoospermic infertile males of Iraqi population
Journal of Genetics, 2020
In human gamete development, the important period is spermatogenesis, which is organized by specific genes on Y chromosome. In some cases, the infertile men have shown microdeletions on Y chromosome, which seemed as if the structural chromosome variance is linked to the reduction of sperm count. This study aimed to determine the frequency and patterns of Y chromosome microdeletions in azoospermia factor (AZF) of Iraqi infertile males. Here, 90 azoospermic infertile males as a study group and 95 normal fertile males as control group were investigated for the microdeletion of AZF loci using numerous sequence-tagged sites. Of these 90 infertile male patients, 43 (47.8%) demonstrated Y chromosome microdeletions, in which AZFb region was the most deleted section in azoospermia patients (33.3%) followed by deletions in the AZFc region (23%), while there were no microdeletion in the AZFa region. The largest microdeletion involved in both AZFb and AZFc was detected in six azoospermic patients (6.7%). The present study demonstrated a high frequency of Y chromosome microdeletions in the infertile Iraqi patients which is not reported previously. The high frequency of deletions may be due to the association of ethnic and genetic factors. PCR-based Y chromosome screening for microdeletions has a potential to be used in infertility clinics for genetic counselling and assisted reproduction.
AZF microdeletions on the Y chromosome of infertile men from Turkey
Annales de Génétique, 2004
Intervals V and VI of Yq11.23 regions contain responsible genes for spermatogenesis, and are named as "azoospermia factor locus" (AZF). Deletions in these genes are thought to be pathogenetically involved in some cases of male infertility associated with azoospermia or oligozoospermia. The aim of this study was to establish the prevalence of microdeletions on the Y chromosome in infertile Turkish males with azoospermia or oligozoospermia. We applied multiplex polymerase chain reaction (PCR) using several sequence-tagged site (STS) primer sets, in order to determine Y chromosome microdeletions. In this study, 61 infertile males were enrolled for the molecular AZF screening program. In this cohort, one infertile male had 46,XX karyotype and the remaining had 46,XY karyotypes. Forty-eight patients had a diagnosis of azoospermia and 13 had oligozoospermia. Microdeletions in AZFa, AZFb and AZFc (DAZ gene) regions were detected in two of the 60 (3.3%) idiopathic infertile males with normal karyotypes and a SRY translocation was determined on 46,XX male. Our findings suggest that genetic screening should be advised to infertile men before starting assisted reproductive treatments.
Y chromosome microdeletion analysis in nonobstructive azoospermia patients from North West of Iran
Jokull
Infertility or sterility is generally described as the inability of a couple to conceive after one year of unprotected coitus (1). Male infertility is a major frequent situation affecting up to half of infertility cases, which comprise approximately 10 to15% of couples (2). This form Aim:The Y-chromosome azoospermic factor (AZF) regions include genes whose specific roles and functions in spermatogenesis have not been completely clarified. Hence, recognition of the association of AZF microdeletions with male infertility has suggestions for the diagnosis, treatment and genetic counseling among infertile patients. Material and Methods:On the basis of cytogenetic evaluation a total of 94 infertile males with nonobstructive azoospermic and aged 24 to 53 years were selected for this study. Molecular AZF screening technique was performed on the genomic DNA from peripheral blood samples. We used Multiplex PCR and four different sets of sequence-tagged sites (STS) for detecting the microdeletions in Y-chromosomal AZF region and theY specific sequences. Results:Among the 94 infertile men, a total of 48 cases (48/94, 51.06%, P< 0.01) were found to have deletions in the regions of AZFb, AZFc and AZFd. Of the 48 azoospermic subjects harbouring Y chromosome microdeletions, twelve had deletions in AZFb, twenty in AZFc, six in AZFb+c, t w o i n AZ F b + c + S R Y, t wo i n AZ F c + d, t w o i n A Z F b + d , t w o i n AZ F b + S R Y a n d two in AZFb+c+d regions. Conclusion:From the results, Y chromosome microdeletions analysis recommend as an important molecular test among infertile males to obtain reliable genetic information before the administration of assisted reproductive techniques and will help decrease the cost and technical difficulty of the procedure.
Defined infertility as incapability of a married couple tᴏ have children for the one-year ᴏf unprotected intercourse. Y chromosomal microdeletions are the second greatest common genetic reason of men sterility. This research aims to find the prevalence of AZF Y chromosome microdeletions in azoospermic and severe oligospermia patients. Further, to evaluate the prevalence types of AZF/c sub-region microdeletions in patients with AZF/c deletion in Iraq. A total of 75 infertile Iraqi males and 25 control were included in this study. The DNA samples were extracted, and they were analysed for AZF microdeletions by utilizing eight sequence-tagged sites through a q/real-time PCR system. After that, partial AZF/c deletion sub-region was investigated using four specific primers. These markers were chosen according to the EAA/ EMGQ recommendations. Out of 75 infertile patients, 46 patients (61.33%) revealed AZF microdeletions in the Y chromosomal with at least one STS deletion for one or more AZF regions. (32.6%) of patients with microdeletions observed in three regions AZFabc. Out of 24 patients who have AZF/c microdeletions (37.5%) were exhibited b2/b4 deletion (complete AZF/c deletions), (58.3%) were showed gr/gr microdeletion (partial AZF/c deletions. and (4.1%) with b2/b4 deletion continues the terminal heterochromatin region. The incidence of classical AZF microdeletions in our study subjects is high. In our study population, gr/gr partial AZF/c microdeletions were higher than b2/b4 complete AZF/c deletion. The mean levels of sex hormones in azoospermic sterile patients with AZF microdeletion were higher than the mean of azoospermic sterile men without deletion of AZF.
JOURNAL OF CLINICAL AND DIAGNOSTIC RESEARCH
Introduction: Azoospermia Factor (AZF) microdeletions in Yq chromosome is one of the most frequent genetic cause associated with failure of spermatogenesis in males with infertility. Aim: To figure out the Yq chromosome microdeletions frequency in infertile men from Gujarat region of India. Materials and Methods: In this study, 141 infertile men with azoospermia (n=41) and oligozoospermia (n=100) were exam ined along with 159 normozoospermic men. Eleven different markers spanning the azoospermia factor region of human Yq chromosome, amplified by sequencetagged site Polymerase Chain Reaction (PCR) to detect the microdeletions. Sperm morphol ogical analysis was done using papanicolau staining method. Results: Thirty four infertile men out of 141 presented Yq chromo some microdeletions. The frequency of AZF microdeletions was 31.71% in azoospermia and 21% in oligozoospermia patients. Only two oligozoospermia patients showed morphological defects. Conclusion: Due to the presence of high frequency of Yq chrom osome microdeletions in Gujarati infertile men, it is imperative to implement the AZF microdeletion screening in such patients as it results in male spermatogenesis dysfunctioning.
Volume 11, Number 4, Jan-Mar 2018
Approximately 15% of couples are infertile with the male factor explaining approximately 50% of the cases. One of the main genetic factors playing a role in male infertility is Y chromosomal microdeletions within the proximal long arm of the Y chromosome (Yq11), named the azoospermia factor (AZF) region. Recent studies have shown there is a potential connection between deletions of the AZF region and recurrent pregnancy loss (RPL). The aim of this study is to examine this association by characterizing AZF microdeletions in two infertile groups: in men with non-obstructive infertility and in men with wives displaying RPL. non-obstructive infertile men, 20 males from couples with RPL and 20 fertile males as controls. Multiplex polymerase chain reaction was used to amplify 19 sequence tagged sites (STS) to detect AZF microdeletions. Differences between the case Only one subject was detected to have Y chromosome microdeletions in SY254, SY157 and SY255 among the 40 men with non-obstructive infertility. No microdeletion was detected in the males with wives displaying RPL and Performing Testing for Y chromosome microdeletions in men with non-obstructive infertility and couples with RPL remains inconclusive in this study.
Male infertility: screening of azoospermia factor (azf) microdeletion in idiopathic infertile men
Journal of Experimental Biology and Agricultural Sciences, 2017
Genetic factors cause about 15% of male infertility and microdeletions of Y chromosome is one of the genetic causes in idiopathic infertile men. Azoospermia factors (AZFa, AZFb, and AZFc) on Yq long arm are most important for spermatogenesis. For analysis of microdeletions in the AZF regions by sequence-tagged-site (STS) PCR is important screening method for infertility. An attempt has been made to evaluate the frequencies of microdeletions of AZFa, AZFb, AZFc in idiopathic cases of azoospermic and oligozoospermic subjects. Total 160 subjects (90 oligozoospermia and 70 azoospermia) and 50 control subjects were analyzed in this study. DNA samples were analyzed for microdeletions of Y chromosome by PCR-screening of 18 STS markers from different locus of the AZFa, AZFb, AZFc on Yq and SRY on Yp. The semen analysis was done and infertile men showing normal karyotype only were included in the study. Plasma follicle stimulating hormone (FSH) and leutinizating hormone concentrations were determined to rule out hormonal abnormality. Out of 160 analyzed cases, 17 (10.6%) subjects showed partial deletion of AZF regions, of which deletion in AZFc region was the most common (58.8%) and it was followed by AZFb and AZFa. The four subjects were shown two or more STS primer deleted sites and overall frequency of Y chromosome microdeletion in our subjects is 10.6%. The sites and sizes of deletions varied among patients. No deletions observed in control subjects. The varied frequencies of Y microdeletions are reported in infertile men in Indian population. From the results of this study it can be suggest that the frequency of deletions may be affected by study sample size, selection criteria of subjects and different geographical region. So, the screening of Y microdeletions is necessary along with the chromosomal analysis in case of infertile men.
Scrreening for microdeletions in human Y chromosome-AZF candidate genes and male infertility
Journal of Cellular and Molecular Medicine, 2003
About 30% of couple infertilities are of male origin, some of them caused by genetic abnormalities of the Y chromosome. Deletions in AZF region can cause severe spermatogenic defects ranging from non-obstructive azoospermia to oligospermia. The intracytoplasmatic sperm injection technique (ICSI) is rapidly becoming a versatile procedure for human assisted reproduction in case of male infertility. The use of ICSI allows Y chromosome defects to be passed from father. The goal of our study is to evaluate the frequency of microdeletions in the long arm of Y chromosome, within the AZF regions, in these cases of infertilities, using molecular genetics techniques. Thirty infertile men with azoospermia or oligozoospermia, determined by spermogram, were studied after exclusion of patients with endocrine or obstructive causes of infertility. Peripheral blood DNA was extracted from each patient, then amplified by multiplex PCR with STS genomic markers from the Y chromosome AZF zones. Each case was checked by multiplex PCR through coamplification with the SRY marker. Three men with microdeletions of the long arm of the Y chromosome were diagnosed among the 30 patients, corresponding to a proportion of 10%. The relatively high proportion of microdeletions found in our population suggest the need for strict patient selection to avoid unnecessary screening for long arm Y chromosome microdeletions. The molecular diagnostics was performed according to the current European Academy of Andrology laboratory guidelines for molecular diagnosis of Y chromosomal microdeletions.
Background: Y chromosomal microdeletion is an important genetic disorder, which may arise due to intrachromosomal recombination between homologous sequences in the male specific region of the human Y chromosome. It is frequently associated with the quantitative reduction of sperm. The screening for Y chromosomal microdeletions has a great clinical value. Objective: To develop a sequence tagged site (STS) based multiplex PCR protocol, which could be specific for the rapid detection of AZF deletions and thereby estimating the frequency of AZF sub deletions in infertile South Indian men. Materials and Methods: In the current study, PCR based Y chromosomal microdeletion screening analysis was performed in 75 men including 30 nonobstructive azoospermic men, 20 severe oligozoospermic, and 25 normozoospermic fertile men (controls) using 15 known STS primer pairs mapped within the AZF locus. Deletion frequency was estimated after successful PCR amplification. Results: We designed and optimized a STS based multiplex PCR protocol, which could be helpful for the clinicians to detect the Y chromosomal deletions rapidly and specifically. In our study, we estimated an overall deletion frequency of 36%. Among these 12 (40%) were azoospermic and 6 (30%) were oligozoospermic. No microdeletions were observed in normozoospermic fertile men. Conclusion: Our Study emphasizes the fact that Y chromosomal microdeletion screening tests are unavoidable in the workup of idiopathic male infertility. Mandatory screening for Y deletions should be done in all azoospermic and severe oligozoospermic patients before undergoing assisted reproductive technology.