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. DESIGN: Experimental study. SETTING: University. PATIENT(S): Men with proven fertility and men with a diagnosis of DFS. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): 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. RESULT(S): 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. CONCLUSION(S): The results show that with effectively designed primers, qPCR may provide an affordable molecular assay to assess male fertility status.
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. DESIGN: Experimental study. SETTING: University. PATIENT(S): Men with proven fertility and men with a diagnosis of DFS. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): 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. RESULT(S): 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. CONCLUSION(S): The results show that with effectively designed primers, qPCR may provide an affordable molecular assay to assess male fertility status.
Authors: Meritxell Jodar; Sellappan Selvaraju; Edward Sendler; Michael P Diamond; Stephen A Krawetz Journal: Hum Reprod Update Date: 2013-07-14 Impact factor: 15.610
Authors: Shihong Mao; Robert J Goodrich; Russ Hauser; Steven M Schrader; Zhen Chen; Stephen A Krawetz Journal: Syst Biol Reprod Med Date: 2013-07-19 Impact factor: 3.061
Authors: Edward Sendler; Graham D Johnson; Shihong Mao; Robert J Goodrich; Michael P Diamond; Russ Hauser; Stephen A Krawetz Journal: Nucleic Acids Res Date: 2013-03-06 Impact factor: 16.971