title: A platform for evaluating sperm RNA biomarkers: dysplasia of the fibrous sheath--testing the concept.
abstract: OBJECTIVE
  To investigate the potential use of correlative microarray-based transcript pairs as candidate markers for male fertility using dysplasia of the fibrous sheath (DFS) as an affected model. It is widely recognized that microarray technology may be limited by cost and that the quality of the transcript remains relatively unknown. To address these issues, we analyzed the stable transcript pairs by qPCR with a systematic primer design process.
  
  
  METHODS
  Experimental study.
  
  
  METHODS
  University.
  
  
  METHODS
  Men with proven fertility and men with a diagnosis of DFS.
  
  
  METHODS
  None.
  
  
  METHODS
  Primer sequences for six genes of interest were designed using Oligo7 and Primer3Plus. Primer specificity was initially assessed in silico by searching the ENSEMBL, University of California Santa Cruz, and National Center for Biotechnology Information databases for nontarget complementary sequences throughout the genome. The ability of transcript pairs to classify samples from males of proven fertility away from DFS was assessed.
  
  
  RESULTS
  In conjunction with identifying four new stable transcript pairs, comparison of the DFS qPCR C(t) correlation coefficients revealed the disruption of four stable fertile sample transcript pairs. This suite of transcript pairs resolves DFS.
  
  
  CONCLUSIONS
  The results show that with effectively designed primers, qPCR may provide an affordable molecular assay to assess male fertility status.
authors.affiliation: Department of Obstetrics and Gynecology, Center for Molecular Medicine and Genetics, C. S. Mott Center, Wayne State University School of Medicine, Detroit, Michigan 48201, USA.
authors.forename: Aletheia
authors.initials: A
authors.lastname: Lima-Souza
authors.forename: Ester
authors.initials: E
authors.lastname: Anton
authors.forename: Shihong
authors.initials: S
authors.lastname: Mao
authors.forename: Won Jin
authors.initials: WJ
authors.lastname: Ho
authors.forename: Stephen A
authors.initials: SA
authors.lastname: Krawetz
mesh_terms: D016022:Case-Control Studies; D017931:DNA Primers; D005298:Fertility / Q000235:genetics*; D020869:Gene Expression Profiling / Q000379:methods; D005819:Genetic Markers; D020022:Genetic Predisposition to Disease; D005820:Genetic Testing; D006801:Humans; D007248:Infertility, Male / Q000175:diagnosis* / Q000235:genetics / Q000473:pathology; D008297:Male; D008824:Michigan; D020411:Oligonucleotide Array Sequence Analysis; D010641:Phenotype; D011237:Predictive Value of Tests; D012313:RNA / Q000032:analysis*; D060888:Real-Time Polymerase Chain Reaction; D015203:Reproducibility of Results; D013082:Sperm Tail / Q000737:chemistry / Q000473:pathology; D013094:Spermatozoa / Q000737:chemistry* / Q000473:pathology*
chemical_list: D017931:DNA Primers; D005819:Genetic Markers; D012313:RNA