Detection and differentiation of norovirus genogroups I and II from clinical stool specimens using real-time PCR.
A real-time RT-PCR assay was designed to detect and differentiate norovirus genogroups I (GI) and II (GII), with primers and probes targeting the nonstructural polyprotein gene. Stool samples (n = 100) submitted for routine testing by the BioFire FilmArray® GI panel were also tested by the norovirus GI/GII real-time PCR assays. When compared to the FilmArray GI panel, the norovirus real-time PCR assay demonstrated a sensitivity of 77.5% (62/80) and specificity of 95% (19/20). Specimens yielding discordant results (n = 19) were tested at two outside laboratories for adjudication. Following discordant resolution, the adjusted sensitivity and specificity of the norovirus real-time PCR assays were 96.9% (63/65) and 100% (35/35), respectively. These results suggest that the real-time PCR assays are able to accurately detect and differentiate norovirus GI/GII from clinical stool specimens. Furthermore, our report highlights a potential issue with the specificity of the BioFire FilmArray® norovirus assay, which warrants additional investigation.
Division of Clinical Microbiology, Mayo Clinic, Rochester, MN, USA 55905.
Poornima
P
Ramanan
Division of Clinical Microbiology, Mayo Clinic, Rochester, MN, USA 55905.
M J
MJ
Espy
Division of Infectious Disease Diagnostics, Northwell Health, Lake Success, New York, NY, USA 11042.
Reeti
R
Khare
Division of Clinical Microbiology, Mayo Clinic, Rochester, MN, USA 55905. Electronic address: binnicker.matthew@mayo.edu.
M J
MJ
Binnicker
D001681:Biological Assay / Q000379:methods; D017250:Caliciviridae Infections; D002454:Cell Differentiation / Q000235:genetics*; D017931:DNA Primers / Q000235:genetics; D005243:Feces / Q000821:virology*; D005759:Gastroenteritis / Q000821:virology; D005838:Genotype; D006801:Humans; D007753:Laboratories; D029322:Norovirus / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
D017931:DNA Primers
Norovirus; PCR