Hepatitis C virus detection in follicular fluid and culture media from HCV+ women, and viral risk during IVF procedures.
BACKGROUND
Hepatitis C virus (HCV) has been detected in sperm, but no data are available on follicular fluid (FF) collected on IVF procedures. The aim of this study was to: (i) search for HCV RNA in FF and in culture media at each step of IVF undergone by HCV(+) women; (ii) investigate the impact of blood contamination of FF induced by vascular injury associated with transvaginal ovarian puncture; (iii) assess risk for the embryo and the impact on the contamination rate of the newborn; and (iv) determine the viral risk presented by these fluids in order to define guidelines for the laboratory.


METHODS
FF from 22 IVF procedures performed in 17 HCV(+) women were classified as either clear, lightly bloody or bloody FF. Oocytes from each FF were washed and incubated in separated fertilization media. At 20 h after puncture (day 1), the fertilized oocytes were washed and transferred to fresh media until embryo transfer. HCV RNA was detected and quantified in FF and media using Cobas Amplicor and Cobas Monitor HCV RNA kits.


RESULTS
HCV RNA was positive in 39 of 44 FF samples, and viral loads increased with blood contamination. At day 1, after rinsing of oocyte-cumulus complexes, only 8 of 44 media were still positive. Viral loads were quantified in 5 of 8 positive media, were below the test sensitivity threshold in 4 of 5 HCV RNA-positive media, and just above it in the fifth medium. The day of transfer HCV RNA was undetectable in all media.


CONCLUSIONS
HCV RNA was detected in 89% of FF irrespective of the degree of blood contamination, and in 25% of the media at day 1. FF must be considered as potentially infected. Blood contamination increases HCV load in the FF. Rinsing oocytes seems significantly to discard the HCV RNA. It is too early to assess the impact of IVF on the contamination rate of HCV mothers' offspring. After counselling, attempting IVF in HCV(+) women is justified. Universal guidelines prevent nosocomial infection, and IVF does not specifically increase the professional risk.
Devaux
A
A
Service d'Histologie-Embryologie-Cytogénétique, Hôpital Bichat-Claude Bernard, AP-HP, Paris, France. aviva.devaux@bch.ap-hop-paris.fr
Soula
V
V
Sifer
C
C
Branger
M
M
Naouri
M
M
Porcher
R
R
Poncelet
C
C
Neuraz
A
A
Alvarez
S
S
Benifla
J L
JL
Madelenat
P
P
Brun-Vezinet
F
F
Feldmann
G
G
D000328:Adult; D001769:Blood / Q000378:metabolism; D003470:Culture Media; D004624:Embryo Transfer; D005260:Female; D005307:Fertilization in Vitro; D015571:Follicular Fluid / Q000378:metabolism / Q000821:virology*; D016174:Hepacivirus / Q000235:genetics / Q000302:isolation & purification*; D006526:Hepatitis C / Q000453:epidemiology / Q000635:transmission / Q000821:virology*; D006801:Humans; D015994:Incidence; D007223:Infant; D018445:Infectious Disease Transmission, Vertical; D009865:Oocytes; D010053:Ovary / Q000601:surgery; D011247:Pregnancy; D011256:Pregnancy Outcome; D018873:Pregnancy Rate; D011677:Punctures / Q000009:adverse effects; D012367:RNA, Viral / Q000032:analysis; D012307:Risk Factors; D007507:Therapeutic Irrigation; D019562:Viral Load
D003470:Culture Media; D012367:RNA, Viral