PMID- 16756977
OWN - NLM
STAT- MEDLINE
DCOM- 20060801
LR  - 20191210
IS  - 1556-5653 (Electronic)
IS  - 0015-0282 (Linking)
VI  - 86
IP  - 1
DP  - 2006 Jul
TI  - Development and evaluation of a qualitative reverse-transcriptase nested 
      polymerase chain reaction protocol for same-day viral validation of human 
      immunodeficiency virus type 1 ribonucleic acid in processed semen.
PG  - 121-8
AB  - OBJECTIVE: To develop a method for same-day validation of processed semen in the 
      setting of assisted reproductive techniques (ART) with patients who are 
      seropositive for human immunodeficiency virus, type 1 (HIV-1). DESIGN: Laboratory 
      experiments. SETTING: University hospital. PATIENT(S): Volunteers who are HIV-1 
      seronegative and seropositive. INTERVENTION(S): Evaluation of the sensitivity of 
      a reverse-transcriptase (RT)-nested polymerase chain reaction (PCR) in HIV-1 
      RNA-positive blood plasma, in artificially infected blood plasma and semen, and 
      in 85 semen samples of 29 HIV-1-seropositive volunteers. Semen was submitted to 
      gradient separation, followed by swim-up. MAIN OUTCOME MEASURE(S): Qualitative 
      detection of HIV-1 RNA in blood plasma and in different parts of semen 
      preparation by using RT-nested PCR, PCR inhibition control by dilution of 
      samples, and an internal control. RESULT(S): The detection limit of our PCR was 
      20 HIV-1 RNA copies per milliliter. Among seropositive patients, RNA was detected 
      in 25% of fresh semen, 36.5% of seminal plasma, 27.5% of gradient supernatants, 
      and 7.1% of final preparations before the migration-sedimentation stage. Positive 
      final preparations were observed in patients who had blood viral loads of 
      >/=20,000 HIV-1 RNA copies per milliliter. Inhibition was present in 17.6% of 
      seminal plasma and in 20% gradient supernatants and in 2 final preparations among 
      69 tested. Among 25 preparations tested after the migration-sedimentation stage, 
      2 were positive (1 patient; 70,000 HIV-1 RNA copies per milliliter). 
      CONCLUSION(S): The RT-nested PCR detects low viral load and allows the validation 
      of semen preparations of HIV-1-seropositive patients for ART on the day of 
      sampling. For this purpose, the validation is performed on spermatozoa that are 
      obtained after gradient separation before swim-up. Inhibition of the PCR must be 
      controlled by using an internal control that is well-designed to explore the 
      detection limit of the method.
FAU - Lesage, Benoit
AU  - Lesage B
AD  - Fertility Clinic, Department of Obstetrics and Gynaecology and Laboratory for 
      Research on Human Reproduction, Campus Erasme, Universite Libre de Bruxelles 
      (ULB), Brussels, Belgium.
FAU - Vannin, Anne-Sophie
AU  - Vannin AS
FAU - Emiliani, Serena
AU  - Emiliani S
FAU - Debaisieux, Laurent
AU  - Debaisieux L
FAU - Englert, Yvon
AU  - Englert Y
FAU - Liesnard, Corinne
AU  - Liesnard C
LA  - eng
PT  - Evaluation Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Validation Study
DEP - 20060606
PL  - United States
TA  - Fertil Steril
JT  - Fertility and sterility
JID - 0372772
RN  - 0 (RNA, Viral)
SB  - IM
MH  - Cells, Cultured
MH  - HIV/*genetics/*isolation & purification
MH  - Humans
MH  - Male
MH  - RNA, Viral/*analysis/genetics
MH  - Reproducibility of Results
MH  - *Reproductive Techniques, Assisted
MH  - Reverse Transcriptase Polymerase Chain Reaction/*methods
MH  - Semen/*virology
MH  - Sensitivity and Specificity
MH  - Viral Load/*methods
EDAT- 2006/06/08 09:00
MHDA- 2006/08/02 09:00
CRDT- 2006/06/08 09:00
PHST- 2005/02/08 00:00 [received]
PHST- 2005/12/12 00:00 [revised]
PHST- 2005/12/12 00:00 [accepted]
PHST- 2006/06/08 09:00 [pubmed]
PHST- 2006/08/02 09:00 [medline]
PHST- 2006/06/08 09:00 [entrez]
AID - S0015-0282(06)00535-8 [pii]
AID - 10.1016/j.fertnstert.2005.12.021 [doi]
PST - ppublish
SO  - Fertil Steril. 2006 Jul;86(1):121-8. doi: 10.1016/j.fertnstert.2005.12.021. Epub 
      2006 Jun 6.