title: Molecular analysis of testis biopsy and semen pellet as complementary methods with histopathological analysis of testis in non-obstructive azoospermia.
abstract: OBJECTIVE
  Non-obstructive azoospermia (NOA) is one the many causes of male infertility (10 %) resulting from testicular failure. Multiple testicular biopsies fail to find mature sperm in at least 50 % of cases Therefore; hunting for sensitive and specific biomarkers of spermatogenesis that could better determine the fertility status in NOA can lead to improved management of male infertility. Therefore, we evaluated sperm production through analyses of germ cell-specific transcripts (DAZ, TSPY1, SPTRX3 and SPTRX1) in semen and testicular biopsies of men with azoospermia.
  
  
  METHODS
  We collected semen (N=83) and testis biopsies (N=31) from men with non-obstructive azoospermia. We later extracted RNA and synthesized cDNA using washed semen precipitate and testicular tissues. We also performed semi-nested PCR with designed specific primers. Using H&E method, an expert pathologist performed the histopathological evaluation. Having categorized the patients into three groups based on histopathological results, we calculated the agreement between molecular results of semen and tissues with histopathological findings for each patient using Kappa statistical test.
  
  
  RESULTS
  Molecular findings of precipitated semen and testicular tissues were in disagreement with histopathological results in most cases. Molecular analysis of testis biopsies showed significant difference (Kappa coefficient=0.009, P value=0.894) with histopathological results; TSPY1, DAZ, SPTRX3 and SPTRX1 were respectively detected in 94 %, 94 %, 17.6 % and 52.9 % of men diagnosed with germ cell aplasia.
  
  
  CONCLUSIONS
  Molecular analysis of semen does not provide sufficient sensitivity and specificity to be used as a screening test at the present time, but it is a useful adjunct to histopathological methods in men with NOA. Spermatid/sperm specific transcripts indicated the possibility to find mature sperm following repeated multiple testicular sperm extraction (TESE) or microdisection TESE (mTESE).
authors.affiliation: Department of Medical Genetics, School of Medicine, Tehran University of Medical Sciences, Poursina Ave, 16 Azar St. Keshavarz BLVD, Tehran, Iran, 1417613151.
authors.forename: Maryam
authors.initials: M
authors.lastname: Eghbali
authors.forename: Mohammad Reza
authors.initials: MR
authors.lastname: Sadeghi
authors.forename: Niknam
authors.initials: N
authors.lastname: Lakpour
authors.forename: Hale
authors.initials: H
authors.lastname: Edalatkhah
authors.forename: Hojjat
authors.initials: H
authors.lastname: Zeraati
authors.forename: Haleh
authors.initials: H
authors.lastname: Soltanghoraee
authors.forename: Mohammad Mehdi
authors.initials: MM
authors.lastname: Akhondi
authors.forename: S Behnam
authors.initials: SB
authors.lastname: Hashemi
authors.forename: Mohammad Hossein
authors.initials: MH
authors.lastname: Modarressi
mesh_terms: D000328:Adult; D053713:Azoospermia / Q000235:genetics* / Q000473:pathology; D001706:Biopsy; D018797:Cell Cycle Proteins / Q000096:biosynthesis; D000075364:Deleted in Azoospermia 1 Protein; D018507:Gene Expression Regulation, Developmental; D006801:Humans; D007248:Infertility, Male / Q000235:genetics / Q000473:pathology*; D008297:Male; D008565:Membrane Proteins / Q000096:biosynthesis; D016601:RNA-Binding Proteins / Q000096:biosynthesis; D012661:Semen / Q000166:cytology; D013091:Spermatogenesis / Q000235:genetics*; D013094:Spermatozoa / Q000473:pathology; D013737:Testis / Q000378:metabolism / Q000473:pathology*; D013879:Thioredoxins / Q000096:biosynthesis
chemical_list: D018797:Cell Cycle Proteins; C095668:DAZ1 protein, human; D000075364:Deleted in Azoospermia 1 Protein; D008565:Membrane Proteins; D016601:RNA-Binding Proteins; C497749:TSPY1 protein, human; C434954:TXNDC2 protein, human; C490377:TXNDC8 protein, human; D013879:Thioredoxins
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