title: One-step real-time PCR assay for detection and quantitation of hepatitis D virus RNA.
abstract: Hepatitis D virus (HDV) is a defective virus which requires hepatitis B virus (HBV) surface antigen (HBsAg) for its assembly. Hepatitis B infected individuals co-infected or superinfected with HDV often present with more severe hepatitis, progress faster to liver disease, and have a higher mortality rate than individuals infected with HBV alone. Currently, there are no commercially available clinical tests for the detection and quantitation of HDV RNA in the United States. A one-step TaqMan quantitative reverse transcription-polymerase chain reaction (qRT-PCR) assay was developed for detection of HDV RNA, designing primers located in the region just downstream from the HDV antigen gene. The assay has the potential to detect all eight HDV genotypes. A quantifiable synthetic RNA control was also developed for use in the determination of HDV RNA titers in clinical samples. The limit of detection of this assay is 7.5×10(2) HDV RNA copies/ml with a dynamic range of six logs. Most clinical specimens tested (40/41) fell within the linear range of the assay. The median HDV RNA titer of the tested specimens was 6.24×10(6) copies/ml, with a range of 8.52×10(3)-1.79×10(9) copies/ml. Out of 132 anti-HDV-positive specimens 41 (31.1%) were positive for HDV RNA.
authors.affiliation: Division of Viral Hepatitis, National Center for HIV, Hepatitis, STD and Tuberculosis Prevention, Centers for Disease Control and Prevention, Atlanta, GA 30333, United States.
authors.forename: Maja
authors.initials: M
authors.lastname: Kodani
authors.forename: Alyssa
authors.initials: A
authors.lastname: Martin
authors.forename: Tonya
authors.initials: T
authors.lastname: Mixson-Hayden
authors.forename: Jan
authors.initials: J
authors.lastname: Drobeniuc
authors.forename: Robert R
authors.initials: RR
authors.lastname: Gish
authors.forename: Saleem
authors.initials: S
authors.lastname: Kamili
mesh_terms: D003699:Hepatitis D / Q000175:diagnosis* / Q000821:virology; D003698:Hepatitis Delta Virus / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D012367:RNA, Viral / Q000235:genetics / Q000302:isolation & purification*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D014481:United States; D019562:Viral Load / Q000379:methods*
chemical_list: D012367:RNA, Viral
keywords: Detection; Hepatitis D virus; One-step; Quantitation; qRT-PCR