title: Complete removal of HIV-1 RNA and proviral DNA from semen by the swim-up method: assisted reproduction technique using spermatozoa free from HIV-1.
abstract: BACKGROUND
  Use of antiretroviral drugs has reduced the mortality rate for HIV infection and many HIV-discordant couples wish to have children. It is possible for an HIV-infected man to father children without risk of HIV transmission if HIV-free spermatozoa can be obtained from his semen.
  
  
  METHODS
  An improved swim-up method was used to collect HIV-free spermatozoa from the semen of HIV-positive males. Diluted semen was layered over a Percoll solution with a continuous density gradient of 30-98%, and then centrifuged. The bottom layer was collected by cutting the end from the tube and the sperm suspension was collected using the swim-up method. Spermatozoa were tested by nested polymerase chain reaction (PCR) for HIV-1 RNA and DNA, with a detection limit of one copy. Spermatozoa were used for assisted reproduction in 43 couples.
  
  
  RESULTS
  HIV-1 RNA and proviral DNA were not detected by nested-PCR assay in all 73 of the collected spermatozoa samples from 52 patients. The HIV-1-negative sperm was used for in vitro fertilization in 12 couples and for intracytoplasmic sperm injection in 31 couples. No detection of HIV-1 RNA or proviral DNA in the culture medium of the fertilized eggs was confirmed again before embryo transfer. Of the 43 female partners, 20 conceived and 27 babies were born. HIV antibodies, HIV RNA and proviral DNA were negative in all of the females and babies.
  
  
  CONCLUSIONS
  HIV-negative spermatozoa could be obtained from semen of HIV-positive men. The method involves no risk of HIV transmission to female partners and their children.
authors.affiliation: Department of Microbiology, Keio University School of Medicine, Tokyo, Japan.
authors.forename: Shingo
authors.initials: S
authors.lastname: Kato
authors.forename: Hideji
authors.initials: H
authors.lastname: Hanabusa
authors.forename: Satoru
authors.initials: S
authors.lastname: Kaneko
authors.forename: Koichi
authors.initials: K
authors.lastname: Takakuwa
authors.forename: Mina
authors.initials: M
authors.lastname: Suzuki
authors.forename: Naoaki
authors.initials: N
authors.lastname: Kuji
authors.forename: Masao
authors.initials: M
authors.lastname: Jinno
authors.forename: Rie
authors.initials: R
authors.lastname: Tanaka
authors.forename: Kenichi
authors.initials: K
authors.lastname: Kojima
authors.forename: Mitsutoshi
authors.initials: M
authors.lastname: Iwashita
authors.forename: Yasunori
authors.initials: Y
authors.lastname: Yoshimura
authors.forename: Kenichi
authors.initials: K
authors.lastname: Tanaka
mesh_terms: D000328:Adult; D023241:Antiretroviral Therapy, Highly Active / Q000379:methods; D004279:DNA, Viral / Q000032:analysis*; D005260:Female; D005307:Fertilization in Vitro / Q000379:methods; D015658:HIV Infections / Q000188:drug therapy / Q000635:transmission / Q000821:virology; D006679:HIV Seropositivity / Q000188:drug therapy / Q000635:transmission / Q000821:virology; D015497:HIV-1; D006801:Humans; D018445:Infectious Disease Transmission, Vertical / Q000517:prevention & control; D008297:Male; D012367:RNA, Viral / Q000032:analysis*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D012661:Semen / Q000821:virology*; D020554:Sperm Injections, Intracytoplasmic / Q000379:methods; D013081:Sperm Motility / Q000502:physiology; D013094:Spermatozoa / Q000821:virology*; D019562:Viral Load; D014771:Virion
chemical_list: D004279:DNA, Viral; D012367:RNA, Viral