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Detection of the <jats:styled-content style=\"fixed-case\">HCV<\/jats:styled-content> genomes in clinical specimens is useful for differential diagnosis, particularly between recurrent <jats:styled-content style=\"fixed-case\">HCV<\/jats:styled-content> infection and acute cellular rejection in transplant specimens. We optimized an <jats:styled-content style=\"fixed-case\">ISH<\/jats:styled-content> protocol that demonstrated sensitivity and specificity for detecting genomic and replicative form of <jats:styled-content style=\"fixed-case\">HCV RNA<\/jats:styled-content> in tissue biopsies. Digoxigenin (<jats:styled-content style=\"fixed-case\">D<\/jats:styled-content>ig)\u2010labelled sense and anti\u2010sense riboprobes were synthesized using a plasmid containing a fragment of the highly conserved <jats:styled-content style=\"fixed-case\">HCV<\/jats:styled-content> noncoding region as a template. The efficiency of the <jats:styled-content style=\"fixed-case\">D<\/jats:styled-content>ig\u2010labelled riboprobes in detecting genomic and replicative\u2010intermediate <jats:styled-content style=\"fixed-case\">HCV RNA<\/jats:styled-content> was analysed in 30 liver biopsies from patients infected or uninfected with <jats:styled-content style=\"fixed-case\">HCV<\/jats:styled-content> in a blinded study. A Huh7 cell line that stably replicates genome\u2010length <jats:styled-content style=\"fixed-case\">HCV RNA<\/jats:styled-content> was developed to be used as a positive control. Negative control riboprobes were used in parallel to evaluate and control for background staining. The anti\u2010sense probe detected <jats:styled-content style=\"fixed-case\">HCV RNA<\/jats:styled-content> in 20\/21 specimens from <jats:styled-content style=\"fixed-case\">HCV<\/jats:styled-content>\u2010infected liver tissues obtained from patients and in 0\/9 samples from patients with non\u2010<jats:styled-content style=\"fixed-case\">HCV<\/jats:styled-content>\u2010related liver diseases, resulting in a sensitivity and specificity of 95% and 100%, respectively. <jats:styled-content style=\"fixed-case\">HCV<\/jats:styled-content> genomic <jats:styled-content style=\"fixed-case\">RNA<\/jats:styled-content> was variably distributed in tissue sections and was located primarily in the perinuclear regions in hepatocytes. Detection of <jats:styled-content style=\"fixed-case\">HCV RNA<\/jats:styled-content> by our optimized <jats:styled-content style=\"fixed-case\">ISH<\/jats:styled-content> protocol appears to be a sensitive and specific method when processing clinical specimens. It may also be revealing when exploring the pathophysiology of <jats:styled-content style=\"fixed-case\">HCV<\/jats:styled-content> infection by verifying the presence of viral genetic material within heptocytes and other cellular elements of diseased liver tissue. This methodology might also evaluate the response to antiviral therapies by demonstrating the absence or alteration of genetic material in clinical specimens from successfully treated patients.<\/jats:p>","DOI":"10.1111\/j.1365-2893.2012.01642.x","type":"journal-article","created":{"date-parts":[[2012,7,30]],"date-time":"2012-07-30T05:36:56Z","timestamp":1343626616000},"page":"183-192","source":"Crossref","is-referenced-by-count":7,"title":["<i>In situ<\/i> hybridization for the detection of hepatitis C virus <scp>RNA<\/scp> in human liver tissue"],"prefix":"10.1111","volume":"20","author":[{"given":"G.","family":"Li","sequence":"first","affiliation":[{"name":"Division of Infectious Diseases Department of Internal Medicine University of Texas Medical Branch Galveston TX USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"K.","family":"Li","sequence":"additional","affiliation":[{"name":"Department of Microbiology, Immunology and Biochemistry University of Tennessee Health Science Center Memphis TN USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"A. 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