Validation of safety procedures for the cryopreservation of semen contaminated with hepatitis C virus in assisted reproductive technology.
BACKGROUND
In France, assisted reproductive technologies involving a hepatitis C virus (HCV)-infected man requires the cryopreservation of potentially infected semen (in order to establish the presence of HCV), hence the need for a safe and secure storage system. We evaluated the safety of high-security straws for the conservation of semen containing HCV RNA under routine conditions.


METHODS
Ionomeric resin (IR) straws were filled with seminal plasma spiked with different concentrations of HCV RNA and sealed using a thermo-solder. After a 4% sodium hypochlorite treatment and/or cryopreservation for 7 days in liquid nitrogen, the outside ends of each straw were rinsed with RNAse-free water.


RESULTS
No HCV RNA could be detected in any of the water samples. Additional samples included the rinsing water from straws sealed by thermo-solder and from the heating wire used to cut the end of straws containing HCV-positive semen. The latter samples were found positive for both HCV RNA and the protamine-2 gene expressed by spermatozoa.


CONCLUSIONS
These results demonstrate the safety of IR straws, the filling system and the thermo-solder for cryopreservation of semen containing HCV in liquid nitrogen. Decontamination of the straw after sealing and the use of disposable scissors to open the straws are strongly recommended.
Maertens
A
A
Laboratoire de Bactériologie-Virologie, GIMAP, University Hospital of Saint-Etienne, 42055, France.
Bourlet
T
T
Plotton
N
N
Pozzetto
B
B
Levy
R
R
D015925:Cryopreservation / Q000295:instrumentation; D016174:Hepacivirus / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D008297:Male; D012367:RNA, Viral / Q000032:analysis; D027724:Reproductive Techniques, Assisted; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012661:Semen / Q000821:virology*; D012662:Semen Preservation; D007507:Therapeutic Irrigation; D014871:Water Microbiology
D012367:RNA, Viral