Comparison of polymerase chain reaction-dependent methods for determining the presence of human immunodeficiency virus and hepatitis C virus in washed sperm

Objectives: 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. Design: Controlled prospective clinical trial. Setting: Private and university-affiliated setting. Patient(s): Thirty-four HIV-positive men attending our center for assisted reproduction with their partners. Intervention(s): 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. Main Outcome Measure(s): Qualitative and quantitative detection of DNA and RNA of HIV and RNA of HCV. Result(s): 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. Conclusion(s): 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.