1. Intended Use
Norovirus real time RT-PCR kit is used for the detection of norovirus in stool or diarrhea samples
by using real time PCR systems.
2. Product Description
Norovirus (NV), one genus of the family of Caliciviridae, is one of the most commonly reported
etiological agents of non-bacterial gastroenteritis in human world-wide. Hospitals, residential
facilities/nursing homes and schools/nurseries were most frequently affected. To prevent further
spreading of the causative agent during a mass outbreak especially in semi closed communities,
such as hospitals and nursing homes, an immediate application of hygiene measures as well as
rapid and sensitive diagnostics are needed. The increasing knowledge of the molecular properties
of caliciviruses led to the development of different assays for the detection of norovirus RNA and
viral antigen. Since enzyme immunoassays were found to be insufficient sensitive and/or
insufficient specific so far RT-PCR assays have become the methods of choice. Due to the high
sequence diversity of human NV which is classified into 3 genogroups (GI, GII, and GIV)
containing at least 7 different genotypes in GI (ie,.Norwalk virus) and 12 genotypes in GII. The
genogroup specific oligonucleotide probes are used in this real-time PCR Kit.
The norovirus real time RT-PCR kit contains a specific ready-to-use system for the detection of the
norovirus Group Ⅰand Group Ⅰusing RT-PCR (Reverse Transcription Polymerase Chain
Reaction) in the real-time PCR system. The master contains a Super Mix for the specific
amplification of the norovirus RNA. The reaction is done in one step real time RT-PCR. The first
step is a reverse transcription (RT), during which the norovirus RNA is transcribed into cDNA.
Afterwards, a thermostable DNA polymerase is used to amplify the specific gene fragments by
means of PCR (polymerase chain reaction). Fluorescence is emitted and measured by the real time
systems´ optical unit during the PCR. The detection of amplified norovirus GⅠand GⅠRNA
fragment is performed in fluorimeter channel 530nm with the fluorescent quencher BHQ1. In
addition, the kit contains a system to identify possible PCR inhibition by measuring the 560nm
fluorescence of the internal control (IC). An external positive control defined as 1×107 copies/ml is
supplied which allow the determination of the gene load. For further information, please refer to
section 9.3 Quantitation.