title: HCV-RNA detection using different PCR methods in sera, cryoglobulins and peripheral blood mononuclear cells of patients with mixed cryoglobulinemia.
abstract: OBJECTIVE
  The hepatitis C virus (HCV) is frequently associated with mixed cryoglobulinemia (MC), and a number of authors have reported the presence of anti-HCV antibodies and HCV-RNA in the blood of MC patients. The presence of the HCV genome in the blood cells of individuals infected by HCV may correlate with the etiopathology of MC. We investigated the presence of HCV-related sequences in the sera, cryoglobulins and peripheral blood mononuclear cells (PBMC) of patients with MC and of individuals with type C chronic active hepatitis (CAH).
  
  
  METHODS
  39 patients with MC, 11 non-cryoglobulinemic HCV-positive individuals with CAH, and 2 anti-HCV negative controls were included in the study. The presence of HCV-RNA was detected by nested RT-PCR and by a commercial kit. The PCR was performed by amplifying the 5'-non coding region (5'-NCR) of HCV.
  
  
  RESULTS
  HCV-RNA was detected in the sera and cryoglobulins of about 90% of the patients; the commercial kit showed a higher sensitivity than nested PCR. One MC patient showed HCV-RNA only in the cryoglobulins. HCV-RNA was present in the PBMC of 14 of the 20 (70%) MC patients analyzed. No differences in serum and PBMC HCV-RNA positivity were found between MC patients and controls.
  
  
  CONCLUSIONS
  Our results confirm the spread of HCV infection among patients with MC. HCV-RNA is present in the serum, cryoglobulins and PBMC of a large proportion of MC patients. The prevalence of HCV-RNA in the PBMC of MC patients and controls did not differ significantly; this may suggest a tropism of HCV for PBMC regardless of the presence of cryoglobulinemia.
authors.affiliation: Clinica delle Malattie Infettive, Università di Milano, Italy.
authors.forename: C
authors.initials: C
authors.lastname: de Maddalena
authors.forename: G
authors.initials: G
authors.lastname: Zehender
authors.forename: A
authors.initials: A
authors.lastname: Bianchi Bosisio
authors.forename: G
authors.initials: G
authors.lastname: Monti
authors.forename: A
authors.initials: A
authors.lastname: Monteverde
authors.forename: F
authors.initials: F
authors.lastname: Invernizzi
authors.forename: M
authors.initials: M
authors.lastname: Galli
mesh_terms: D001483:Base Sequence; D003449:Cryoglobulinemia / Q000097:blood / Q000821:virology*; D003450:Cryoglobulins / Q000032:analysis*; D017931:DNA Primers; D005260:Female; D016174:Hepacivirus / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D009000:Monocytes / Q000821:virology*; D016133:Polymerase Chain Reaction / Q000379:methods; D012367:RNA, Viral / Q000032:analysis*
chemical_list: D003450:Cryoglobulins; D017931:DNA Primers; D012367:RNA, Viral