A nucleic acid sequence-based amplification assay for real-time detection of norovirus genogroup II.
OBJECTIVE
To use molecular beacon based nucleic acid sequence-based amplification (NASBA) to develop a rapid, sensitive, specific detection method for norovirus (NV) genogroupII (GII).


RESULTS
A method to detect NV GII from environmental samples using real-time NASBA was developed. This method was routinely sensitive to 100 copies of target RNA and intermittent amplification occurred with as few as 10 copies. Quantitative estimates of viral load were possible over at least four orders of magnitude.


CONCLUSIONS
The NASBA method described here is a reliable and sensitive assay for the detection of NV. This method has the potential to be linked to a handheld NASBA device that would make this real-time assay a portable and inexpensive alternative to bench-top, lab-based assays.


CONCLUSIONS
The development of the real-time NASBA assay described here has resulted in a simple, rapid (<1 h), convenient testing format for NV. To our knowledge, this is the first example of a molecular beacon based NASBA assay for NV.
Patterson
S S
SS
College of Marine Science, University of South Florida, St Petersburg, FL 33701, USA.
Smith
M W
MW
Casper
E T
ET
Huffman
D
D
Stark
L
L
Fries
D
D
Paul
J H
JH
D001483:Base Sequence; D017250:Caliciviridae Infections / Q000175:diagnosis*; D015342:DNA Probes / Q000235:genetics; D004279:DNA, Viral / Q000032:analysis*; D004783:Environmental Microbiology; D005243:Feces / Q000821:virology; D050456:Fomites / Q000821:virology; D005759:Gastroenteritis / Q000821:virology*; D005838:Genotype; D006801:Humans; D008969:Molecular Sequence Data; D029322:Norovirus / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D021161:Self-Sustained Sequence Replication / Q000379:methods*; D019562:Viral Load
D015342:DNA Probes; D004279:DNA, Viral