title: Detection of HIV and HCV RNA in semen from Brazilian coinfected men using multiplex PCR before and after semen washing.
abstract: BACKGROUND
  Prolonged survival of patients under HAART has resulted in new demands for assisted reproductive technologies. HIV serodiscordant couples wish to make use of assisted reproduction techniques in order to avoid viral transmission to the partner or to the newborn. It is therefore essential to test the effectiveness of techniques aimed at reducing HIV and HCV loads in infected semen using molecular biology tests.
  
  
  METHODS
  After seminal analysis, semen samples from 20 coinfected patients were submitted to cell fractioning and isolation of motile spermatozoa by density gradient centrifugation and swim-up. HIV and HCV RNA detection tests were performed with RNA obtained from sperm, seminal plasma and total semen.
  
  
  RESULTS
  In pre-washing semen, HIV RNA was detected in 100% of total semen samples, whereas HCV RNA was concomitantly amplified in only one specimen. Neither HIV nor HCV were detected either in the swim-up or in the post-washing semen fractions.
  
  
  CONCLUSIONS
  Reduction of HIV and/or HCV shedding in semen by density gradient centrifugation followed by swim-up is an efficient method. These findings lead us to believe that, although semen is rarely found to contain HCV, semen processing is highly beneficial for HIV/HCV coinfected individuals.
authors.affiliation: Laboratório de Virologia, Instituto de Medicina Tropical, Faculdade de Medicina, Universidade de São Paulo, São Paulo, SP, Brazil. ccanto@usp.br
authors.forename: Cynthia Liliane Motta do
authors.initials: CL
authors.lastname: Canto
authors.forename: Aluisio C
authors.initials: AC
authors.lastname: Segurado
authors.forename: Cláudio
authors.initials: C
authors.lastname: Pannuti
authors.forename: Agnaldo
authors.initials: A
authors.lastname: Cedenho
authors.forename: Miguel
authors.initials: M
authors.lastname: Srougi
authors.forename: Deborah
authors.initials: D
authors.lastname: Spaine
authors.forename: Silvana
authors.initials: S
authors.lastname: Fernandes
authors.forename: Nadily
authors.initials: N
authors.lastname: Carretiero
authors.forename: Maria Carolina
authors.initials: MC
authors.lastname: Bernal
authors.forename: José Eduardo
authors.initials: JE
authors.lastname: Levi
mesh_terms: D000328:Adult; D002469:Cell Separation; D002499:Centrifugation, Density Gradient; D016679:Genome, Viral / Q000235:genetics; D006678:HIV / Q000235:genetics / Q000302:isolation & purification*; D015658:HIV Infections / Q000517:prevention & control / Q000821:virology; D016174:Hepacivirus / Q000235:genetics / Q000302:isolation & purification*; D006526:Hepatitis C / Q000517:prevention & control / Q000821:virology; D006801:Humans; D008297:Male; D008875:Middle Aged; D012367:RNA, Viral / Q000032:analysis*; D027724:Reproductive Techniques, Assisted; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012661:Semen / Q000821:virology*; D013094:Spermatozoa / Q000821:virology*; D019562:Viral Load
chemical_list: D012367:RNA, Viral