title: HIV-1 detection by nested PCR and viral culture in fresh or cryopreserved postmortem skin: potential implications for skin handling and allografting.
abstract: OBJECTIVE
  To date, the risk relating to the handling or allografting of human immunodeficiency virus type 1 (HIV-1) infected postmortem skin remains hypothetical. While blood screening for HIV antibodies is still the key safety procedure to detect HIV infected cadavers, false negative results are a concern. Conversely, false positive results may hamper the collection of skin allografts. Accordingly, viral culture was used to clarify skin infectivity and the nested polymerase chain reaction (PCR) was used to assess the reliability of skin PCR testing.
  
  
  METHODS
  Viral culture and nested PCR performed with gag and pol specific primers were investigated in cadaveric skin and blood from 12 HIV-1 infected patients. Samples were collected repeatedly between one and five days in seven patients. In most cases, analyses were performed on triplicate skin samples: fresh (n = 26); cryopreserved in 5% dimethylsulphoxide (n = 21), or cryopreserved in 30% glycerol (n = 26).
  
  
  RESULTS
  HIV was isolated in two of 26 cultures of fresh skin specimens (8%), seven of 47 cryopreserved skin specimens (15%), and eight of 26 blood specimens (31%). The nested PCR detected HIV-1 in all skin samples (n = 73), regardless of the postmortem interval or cryopreservation. In blood, a positive signal was found in eight of 12 patients but two of them had discordant results on successive samples.
  
  
  CONCLUSIONS
  These data suggest that nested PCR on postmortem skin samples can detect HIV more reliably than on blood. They also demonstrate the potential viral infectivity of fresh or stored skin postmortem samples in HIV infected patients. They underscore the need for caution during the handling of skin tissue from HIV infected cadavers and confirm the potential risk related to accidental allografting of HIV contaminated skin.
authors.affiliation: Department of Clinical Molecular Biology, Saint Luc Hospital, Catholic University of Louvain, Brussels, Belgium.
authors.forename: J L
authors.initials: JL
authors.lastname: Gala
authors.forename: A T
authors.initials: AT
authors.lastname: Vandenbroucke
authors.forename: B
authors.initials: B
authors.lastname: Vandercam
authors.forename: J P
authors.initials: JP
authors.lastname: Pirnay
authors.forename: N
authors.initials: N
authors.lastname: Delferrière
authors.forename: G
authors.initials: G
authors.lastname: Burtonboy
mesh_terms: D000163:Acquired Immunodeficiency Syndrome / Q000821:virology*; D002102:Cadaver; D016022:Case-Control Studies; D015925:Cryopreservation; D005188:False Negative Reactions; D015497:HIV-1 / Q000302:isolation & purification*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D012867:Skin / Q000821:virology*; D016038:Skin Transplantation; D014184:Transplantation, Homologous