214: Identification of hepatitis C virus in cervico-vaginal secretions of infected women

INFECTED WOMEN. KIRAN TAM TAM, ANTONIO SISON, OMAR BAGASRA, HIMA TAM TAM, DAVID JASPAN, Clinica Campesina Family Health Services, Obstetrics & Gynecology, Denver, Colorado, Robert Wood Johnson University Hospital Hamilton, Obstetrics & Gynecology, Hamilton, New Jersey, South Carolina Center for Biotechnology, Department of Biology, Orangeburg, South Carolina, North Shore University Hospital, Maternal Fetal Medicine, Manhasset, New York, Albert Einstein Medical Center, Obstetrics & Gynecology, Philadelphia, Pennsylvania OBJECTIVE: To provide evidence for heterosexual and perinatal (non-transplacental) transmission by demonstrating the presence of HCV in cervico-vaginal secretions of infected women. STUDY DESIGN: Six HCV infected women in the Albert Einstein Health Care Network were recruited during the period 9/00-5/01. Exclusion criteria included the following 1) patient is menstruating, 2) patient is medically unstable to obtain the necessary specimens, 3) patient is in active labor, 4) patient with premature rupture of the membranes, and 5) patient is obstetrically unstable to obtain necessary specimens. Patients who were co-infected with human immunodeficiency virus (HIV) or Hepatitis B virus were not excluded from the study. 20 cc of cervicovaginal lavage (CVL) was obtained from each patient. CVL specimens were divided into cellular and acellular components. RNA was isolated by a modified guanidinium thiocyanate phenol-chloroform technique with a commercially available kit (Ultraspec 2, Biotecx Laboratories, Houston, TX). RNA was subjected to reverse-transcriptase polymerase chain reaction (RT-PCR). Analysis of HCV quasispecies was conducted on the stable 5’ untranslated region. HCV sequences were compared by the single-stranded conformation polymorphism (SSCP) assay. The SSCP products were sequenced and the secondary RNA structures identified by the University of Rochester=s RNA structure program. Dual RNA/DNA in situ PCR at the single cell level was also used in identifying HCV. RESULTS: All six women were HIV positive and were not pregnant. In all six cases HCV was isolated in both the cellular and acellular components of the cervicovaginal lavage specimen.