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Resume Objectifs. – La possibilite de prendre en charge en assistance medicale a la procreation (AMP) des couples seropositifs, vis-a-vis du VIH-1 en particulier, a reactive la reflexion sur l'aspect securitaire de la conservation de tels gametes et des embryons issus de ceux-ci dans des cuves d'azote liquide. Ainsi nous avons voulu tester celui-ci vis-a-vis de la cryoconservation. Patients et methodes. – Nous avons evalue la securite de trois types de paillettes, polyvinyl chloride (PVC), polyethylene terephtalate glycol (PETG), et resine ionomere (RI) dites de « haute securite », contenant du VIH-1 dans les conditions de cryoconservation. La recherche de la contamination par le VIH a ete realisee par RT-PCR puis nested PCR. Resultats. – Cette etude, menee dans les conditions de cryoconservation, montre que l'extremite libre obturee des paillettes PVC et PETG n'est pas securitaire. Le systeme de fermeture par ultrasons semblerait etre le maillon faible pour l'etancheite parfaite des paillettes. A l'inverse, les paillettes RI pour leurs deux extremites sont securitaires vis-a-vis du VIH dans le cadre de leur utilisation en AMP. Conclusion. – Le systeme de fermeture par ultrasons pourrait constituer un risque concernant l'impermeabilite des paillettes. Les paillettes RI, thermosoudees apparaissent securitaires vis-a-vis du VIH dans les conditions de cryoconservation.

The aim of this preliminary experimental study was to test the stability of cryopreservation straws to human immunodeficiency virus-1 (HIV-1). Three kinds of straws were tested: four polyvinyl chloride (PVC), four polyethylene terephthalate glycol (PETG) and 20 high-security ionomeric resin (IR). The PVC and PETG straws were sealed ultrasonically, and the IR straw by thermosoldering. Each sealed straw was cut in half to produce two demi-straws and then filled with 100 microl of HIV-1-containing supernatant (reverse transcriptase activity: 15 000 c.p.m./50 microl). The unsealed cotton end of PVC and PETG straws and the two halves of the IR straws (cotton and plastic plug ends) were tested. Each demi-straw was two- thirds submerged in RPMI medium at 37 degrees C, and RPMI samples were withdrawn on days 3, 7 and 11. Viral RNA was extracted from the medium and then amplified by reverse transcriptase-polymerase chain reaction (RT-PCR) followed by nested PCR using primers specific to HIV-1 protease. On day 7, no HIV-1 RNA was detected in any of the different samples of medium that had surrounded the unsealed PVC and PETG straws with cotton ends, but three IR specimens were positive. On day 11, PVC and PETG remained negative but HIV-1 RNA was detected in RPMI samples for two more IR demi-straws (n = 5). In conclusion, under these experimental conditions (at 37 degrees C), the unsealed cotton end PVC, PETG and thermosoldered cotton end IR demi-straws appeared to be safe for HIV-1, while IR straws, sealed or unsealed with a plastic plug and with unsealed cotton ends, leaked.

Background The possibility of assisted reproductive technology (ART) for couples carrying viruses, especially HIV-1, necessitates consideration of the safety of cryopreserving human gametes or embryos in liquid nitrogen tanks. Following our evaluation of the safety of three kinds of straws containing HIV-1 at 37 degrees C, we have now examined the HIV-1 imperviousness of the same straws under cryopreservation conditions. Methods Polyvinyl chloride (PVC), polyethylene terephthalate glycol (PETG) and high-security ionomeric resin (IR) straws (24 each) were tested. Each straw was filled with 100 microl of HIV-1-containing supernatant [reverse transcriptase (RT) activity: 15 000 c.p.m./50 microl]. Then PVC and PETG straws were sealed ultrasonically only at their free-end, and IR straws were thermosoldered at both ends. Each straw was put in a 15 ml Falcon tube which was capped and submerged in a liquid-nitrogen tank for 7 days. After bleach decontamination or not, the outside of each end of the straw was rinsed with RPMI medium (1 ml) before cryopreservation and after thawing. Viral RNA was extracted from the medium and then amplified by RT-polymerase chain reaction (PCR) followed by nested-PCR using HIV-1 protease-specific primers. Results HIV-1 RNA was detected in some PVC and PETG rinse media, probably resulting from splashing during ultrasonic sealing, but not in the rinse media of thermosoldered IR straws. Conclusion Under cryopreservation conditions, IR straws would appear to be safe for HIV-1 storage in ART. For PVC and PETG straws, as highlighted in this study, the ultrasonic sealing could be the weak safety link.