title: Novel approach for quantification of hepatitis C virus in liver cirrhosis using real-time reverse transcriptase PCR.
abstract: BACKGROUND
  Hepatitis C virus (HCV) infects nearly 3% of the population worldwide and is a major cause of acute and chronic infections leading to fibrosis, cirrhosis, and hepatocellular carcinoma. Current laboratory diagnosis of HCV is based on specific antibody detection (anti-hepatitis C virus (anti-HCV)) in serum. As HCV replicates in the liver cells, detection and localization of HCV RNA in liver tissue are vital for diagnosis.
  
  
  METHODS
  Ten biopsy samples diagnosed for cryptogenic liver cirrhosis, negative for the presence of anti-HCV and serum HCV RNA, were studied for analyzing presence of viral nucleic acid in liver tissues. Qualitative screening for HCV was done through ELISA while the nucleic acid analysis was performed through COBAS Amplicor. Detection of HCV RNA in liver tissue biopsies was performed following standard protocol of HCV detection kit (Shenzhen PG Biotech) with modifications using Light Cycler 2.0 (minimum detection limit 10 copies/ml).
  
  
  RESULTS
  Quantitative detection in liver biopsies following the modified method showed the presence of HCV RNA in three samples out of the ten studied.
  
  
  CONCLUSIONS
  The results indicate that using Light Cycler 2.0, following the modified technique described, constitutes a reliable method of quantitative detection and localization of HCV in tissue in "serosilent" HCV infection.
authors: Department of Surgical Gastroenterology, Bhopal Memorial Hospital & Research Center, Karond, Raisen Bypass Road, Bhopal, 462 038, Madhya Pradesh, India. maudarji@yahoo.com
authors: Kewal Krishan
authors: KK
authors: Maudar
authors: Puneet
authors: P
authors: Gandhi
authors: Pradyumna Kumar
authors: PK
authors: Mishra
authors: Subodh
authors: S
authors: Varshney
authors: Ramprakash
authors: R
authors: Punde
authors: Arpit
authors: A
authors: Bhargav
mesh_terms: D000328:Adult; D001706:Biopsy; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005838:Genotype; D016174:Hepacivirus / Q000235:genetics / Q000302:isolation & purification*; D018937:Hepatitis C Antibodies / Q000097:blood; D018936:Hepatitis C Antigens / Q000276:immunology; D019698:Hepatitis C, Chronic / Q000175:diagnosis / Q000821:virology; D006801:Humans; D008099:Liver / Q000737:chemistry / Q000473:pathology* / Q000821:virology*; D008103:Liver Cirrhosis / Q000473:pathology* / Q000821:virology*; D008297:Male; D008875:Middle Aged; D012313:RNA / Q000032:analysis*; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012698:Serologic Tests
chemical_list: D018937:Hepatitis C Antibodies; D018936:Hepatitis C Antigens; D012313:RNA