title: Rapid and sensitive approach to simultaneous detection of genomes of hepatitis A, B, C, D and E viruses.
abstract: BACKGROUND
  Five viruses have been etiologically associated with viral hepatitis. Nucleic acid testing (NAT) remains the gold standard for diagnosis of viremic stages of infection. NAT methodologies have been developed for all hepatitis viruses; however, a NAT-based assay that can simultaneously detect all five viruses is not available.
  
  
  OBJECTIVE
  We designed TaqMan card-based assays for detection of HAV RNA, HBV DNA, HCV RNA, HDV RNA and HEV RNA.
  
  
  METHODS
  The performances of individual assays were evaluated on TaqMan Array Cards (TAC) for detecting five viral genomes simultaneously. Sensitivity and specificity were determined by testing 329 NAT-tested clinical specimens.
  
  
  RESULTS
  All NAT-positive samples for HCV (n = 32), HDV (n = 28) and HEV (n = 14) were also found positive in TAC (sensitivity, 100%). Forty-three of 46 HAV-NAT positive samples were also positive in TAC (sensitivity, 94%), while 36 of 39 HBV-NAT positive samples were positive (sensitivity, 92%). No false-positives were detected for HBV (n = 32), HCV (n = 36), HDV (n = 30), and HEV (n = 31) NAT-negative samples (specificity 100%), while 38 of 41 HAV-NAT negative samples were negative by TAC (specificity 93%).
  
  
  CONCLUSIONS
  TAC assay was concordant with corresponding individual NATs for hepatitis A-E viral genomes and can be used for their detection simultaneously. The TAC assay has potential for use in hepatitis surveillance, for screening of donor specimens and in outbreak situations. Wider availability of TAC-ready assays may allow for customized assays, for improving acute jaundice surveillance and for other purposes for which there is need to identify multiple pathogens rapidly.
authors.affiliation: Division of Viral Hepatitis, National Center for HIV, Hepatitis, STD and TB Prevention, Centers for Disease Control and Prevention, Atlanta, GA 30293, United States. Electronic address: mkodani@cdc.gov.
authors.forename: Maja
authors.initials: M
authors.lastname: Kodani
authors.affiliation: Division of Viral Hepatitis, National Center for HIV, Hepatitis, STD and TB Prevention, Centers for Disease Control and Prevention, Atlanta, GA 30293, United States.
authors.forename: Tonya
authors.initials: T
authors.lastname: Mixson-Hayden
authors.affiliation: Division of Viral Hepatitis, National Center for HIV, Hepatitis, STD and TB Prevention, Centers for Disease Control and Prevention, Atlanta, GA 30293, United States.
authors.forename: Jan
authors.initials: J
authors.lastname: Drobeniuc
authors.affiliation: Division of Viral Hepatitis, National Center for HIV, Hepatitis, STD and TB Prevention, Centers for Disease Control and Prevention, Atlanta, GA 30293, United States.
authors.forename: Saleem
authors.initials: S
authors.lastname: Kamili
mesh_terms: D006518:Hepatitis Viruses / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D006525:Hepatitis, Viral, Human / Q000175:diagnosis* / Q000821:virology*; D006801:Humans; D044085:Microfluidics / Q000379:methods*; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods; D012680:Sensitivity and Specificity; D013997:Time Factors
keywords: Hepatitis; Microfluidics; Real-time PCR; Simultaneous pathogen detection