Detection of negative-sense RNA in packaged hepatitis E virions by use of an improved strand-specific reverse transcription-PCR method.
Current hepatitis E virus (HEV) negative-sense RNA detection assays have the drawback of false positivity. cDNA synthesis using tag-based primer and Superscript RT-III followed by exonuclease I treatment increased the specificity. Assays could detect as few as 10 copies of negative-sense RNA and could be used in detecting low levels of HEV replication in cells. Virus particles in stool samples of hepatitis E patients showed encapsidation of negative-sense RNA along with HEV genomic RNA.
Hepatitis Division, National Institute of Virology, Pashan, Pune, India.
Subhashis N
SN
Chatterjee
Pradip B
PB
Devhare
Kavita S
KS
Lole
D000208:Acute Disease; D005189:False Positive Reactions; D005243:Feces / Q000821:virology; D016679:Genome, Viral; D016751:Hepatitis E / Q000175:diagnosis* / Q000821:virology; D016752:Hepatitis E virus / Q000235:genetics* / Q000502:physiology; D006801:Humans; D016372:RNA, Antisense / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D014771:Virion / Q000235:genetics*; D014779:Virus Replication
D016372:RNA, Antisense