Buoyant density and sedimentation dynamics of HIV-1 in two density-gradient media for semen processing

Fertil Steril. 2008 Nov;90(5):1983-7. doi: 10.1016/j.fertnstert.2007.09.025. Epub 2007 Dec 31.

Abstract

Objective: To compare buoyant density and sedimentation kinetics of human immunodeficiency virus 1 (HIV-1) in two sperm-washing media, Percoll and Pureception.

Design: Laboratory study.

Setting: University hospital.

Patient(s): None.

Intervention(s): Buoyant density and sedimentation kinetics of HIV-1 particles (MOLT-4/LAI strain) were measured in Percoll and Pureception using isopyknic ultracentrifugation and continuous-density-gradient centrifugation.

Main outcome measure(s): The HIV-1 particles were detected and semiquantified using a reverse transcription polymerase chain reaction (RT-PCR) for HIV-1 RNA.

Result(s): Calculated buoyant density of HIV-1 was approximately 1.042 in both media in isopyknic centrifugation. However, most HIV-1 particles were found in fractions with specific gravity less than 1.04 in both media, even after 40 minutes of density-gradient centrifugation at 1,600 g. Small viral accumulations were observed at the bottom of the tube in Pureception density gradients.

Conclusion(s): Although we found very high efficiency of HIV-1 removal using density-gradient centrifugation, a minute quantity of virus was found at the bottom of the gradient tube when Pureception was used as the medium.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Centrifugation, Density Gradient*
  • Centrifugation, Isopycnic*
  • HIV-1 / genetics
  • HIV-1 / isolation & purification*
  • Humans
  • Kinetics
  • Male
  • Povidone
  • RNA, Viral / isolation & purification
  • Reproducibility of Results
  • Reverse Transcriptase Polymerase Chain Reaction
  • Semen / virology*
  • Silicon Dioxide
  • Specimen Handling / methods*
  • Viral Load

Substances

  • RNA, Viral
  • Percoll
  • Silicon Dioxide
  • Povidone