Varicocele Clinical Trial
Official title:
Seminal TEX101 as a Predictor of Recovery of Spermatogenesis in Azoospermic Men With Palpable Varicocele
The aim of the study is to assess if seminal TEX 101 can predict the restoration of spermatogenesis and appearance of sperms in the ejaculate after micro-surgical varicocelectomy among azoospermic patients.
Male patients (40) cases attending the outpatient clinic of Andrology at KASR EL-AINI
university hospitals, all patients presented with azoospermia, age:18-43 years, complaining
mainly of infertility or accidently discovered azoospermia during premarital check-up or
chronic testicular pain caused by longstanding varicocele. Infertile Non Obstructive
Azoospermia (NOA) (n=40) subjected to varicocelectomy and patients evaluated 3 months after
varicocelectomy by semen analysis, then according to results population divided into 2
groups: Group A (Responders): Patients with positive semen analysis for
sperms 3 months after varicocelectomy.
Group B (Non Responders): Patients with no sperms in semen analysis
in 3 and/or 6 months after varicocelectomy.
Method of Estimation of human TEX101 in semen:
A 1 ml collected semen samples were centrifuged for 20 minutes at 1000xg at 4 o C, then
supernatant of each sample was collected for estimation of TEX101. The human TEX101 (Testis
expressed 101) was assayed by commercially available ELISA kit supplied by Wuhan Fine Biotech
Co., Ltd. China.
Principle of the procedure:
This kit was based on sandwich enzyme-linked immune-sorbent assay technique. Anti- TEX101
antibody was pre-coated onto 96-well plate. The biotin conjugated antibody was used as
detection antibodies. The standards, test samples and biotin conjugated detection antibody
were added to the wells subsequently, and washed with wash buffer.
HRP-Streptavidin was added and unbound conjugates were washed away with wash buffer. TMB
substrates were used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce
a blue color product that changed into yellow after adding acidic stop solution. The
intensity of yellow is proportional to the target amount of sample captured in plate.
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