Letters to the EditorFalse detection of negative-strand hepatitis C virus RNA
References (9)
- et al.
Detection of plus and minus HCV RNA in normal liver of anti-HCV-positive patients
Lancet
(1993) - et al.
Identification and characterization of type 2 dengue virus replicative intermediate and replicative form RNAs
Virology
(1981) - et al.
PCR and detection of negative HCV RNA strands
Hepatology
(1993) - et al.
Failure to detect hepatitis C virus genome in human secretions with the polymerase chain reaction
Hepatology
(1991)
Cited by (68)
Viral genome imaging of hepatitis C virus to probe heterogeneous viral infection and responses to antiviral therapies
2016, VirologyCitation Excerpt :Infected cells also cluster bimodally into subpopulations bearing low or high strand counts (Fig. S2). In order to cross-validate our vRNA measurements with an independent assay, we developed a new PCR assay that improves our capacity to discriminate between positive and negative strand vRNAs, since the application of previously published methods yield NPSR values for HCV that range widely from 1:10 to 1:1000, and these readings have been confounded by both interference from the opposite strand, and detection of incomplete vRNAs (McGuinness et al., 1994; Revie and Salahuddin, 2011; Agnello et al., 1998; Komurian-Pradel et al., 2004; Lanford et al., 1995; Lohmann et al., 1999). In this updated assay, total RNA is poly-A tailed, after which a tag oligo-dT anchored primer is used to reverse transcribe from the nascent tail (Fig. S3A); the resulting complementary DNA (cDNA) is then used for qPCR, employing a primer for the tag sequence and an HCV-specific primer for the 3′ end of the target vRNA.
Association of chronic hepatitis C with recurrent brief depression
2012, Journal of Affective DisordersCitation Excerpt :Although hepatocytes are the primary site of HCV replication in the body, several studies have identified viral sequences in other anatomic sites, such as blood cells (Lerat et al., 1996), lymphoid tissue (Laskus et al., 2000), and the central nervous system (Fishman et al., 2008; Forton et al., 2004; Laskus et al., 2002; Radkowski et al., 2002), leading to the postulate that HCV may replicate in extrahepatic sites. However, this issue has remained controversial because of the questionable specificity of the tests used to determine the presence of active HCV replication (Farci, 2011), in particular assays for the detection of negative-strand HCV RNA (Lanford et al., 1995; Laskus et al., 1997; McGuinness et al., 1994). Other difficulties are related to the correct preservation of HCV RNA, especially in post-mortem tissues, such as brain tissue.
Hepatitis C virus lymphotropism: lessons from a decade of studies
2007, Digestive and Liver DiseaseIntracellular hepatitis C virus RNA-dependent RNA polymerase activity
2003, Journal of Virological MethodsDifferential distribution and internal translation efficiency of hepatitis C virus quasispecies present in dendritic and liver cells
2003, BloodCitation Excerpt :In addition, tissue compartmentalization of the HCV genome has been described during HCV infection7,8and coinfection with HIV.8 Finally, the detection of HCV RNA-negative strand as the theoretical intermediate of replication, although somewhat controversial,9,10has been well documented in PBMCs. Indeed, negative- strand RNA has been reported in mononuclear cells such as B and T lymphocytes and monocytes,11-15in polynuclear lymphocytes,16,17and even in hematopoietic progenitor cells.18