title: Comparison of polymerase chain reaction-dependent methods for determining the presence of human immunodeficiency virus and hepatitis C virus in washed sperm.
abstract: OBJECTIVE
  To evaluate the effectiveness of sperm washes by double capacitation in patients with human immunodeficiency virus (HIV) and hepatitis C virus (HCV) and to compare between polymerase chain reaction (PCR)-dependent methods to determine viral presence after the procedure.
  
  
  METHODS
  Controlled prospective clinical trial.
  
  
  METHODS
  Private and university-affiliated setting.
  
  
  METHODS
  Thirty-four HIV-positive men attending our center for assisted reproduction with their partners.
  
  
  METHODS
  Forty-one semen samples from seropositive males were obtained and washed through density gradients and swim-up and analyzed for the presence of HIV and HCV with either nested or one-round PCR. Qualitative and quantitative detection of DNA and RNA of HIV and RNA of HCV.
  
  
  RESULTS
  Of all the semen samples that were analyzed after washing, seven out of 41 samples tested positive for one or both viruses by nested PCR. The confirmation of these results with the currently used commercial methods of quantitative PCR (one-round PCR) resulted in all negatives.
  
  
  CONCLUSIONS
  Sperm wash with nested PCR is the appropriate method to use in the assisted reproduction techniques that are offered to serodiscordant couples. The detection limits exhibited by one-round PCR do not offer a sufficient guarantee that transmission of all viral particles will be avoided since both viruses can be present in amounts lower than the detection threshold of this technique.
authors.affiliation: Instituto Valenciano de Infertilidad Valencia (IVI), Valencia, Spain. andrologiavalencia@ivi.es
authors.forename: Marcos
authors.initials: M
authors.lastname: Meseguer
authors.forename: Nicolás
authors.initials: N
authors.lastname: Garrido
authors.forename: Concha
authors.initials: C
authors.lastname: Gimeno
authors.forename: José
authors.initials: J
authors.lastname: Remohí
authors.forename: Carlos
authors.initials: C
authors.lastname: Simón
authors.forename: Antonio
authors.initials: A
authors.lastname: Pellicer
mesh_terms: D000328:Adult; D006678:HIV / Q000302:isolation & purification*; D006679:HIV Seropositivity / Q000821:virology; D016174:Hepacivirus / Q000302:isolation & purification*; D006801:Humans; D008297:Male; D016133:Polymerase Chain Reaction; D011446:Prospective Studies; D013094:Spermatozoa / Q000821:virology*; D007507:Therapeutic Irrigation / Q000592:standards*; D019562:Viral Load