Norovirus – real-time PCR detection
The SureFast ® Norovirus PLUS Kit is a real-time PCR based detection kit for Noroviruses of genogroup I and II. Viral RNA is transcribed to cDNA by using a reverse transcriptase and is subsequently amplified by PCR. The detection is performed in a “One Step RT-qPCR” system: Reverse transcription and subsequent PCR are performed in only one reaction tube. For detection in food matrices and contaminated surfaces classical methods like Enzyme Linked Immunosorbant Assay (ELISA) are not suitable. This is due to low infection dose that requires detection of < 1000 virus particles. Hence, a main advantage of real-time RT-qPCR is the rapid and sensitive detection of small amounts of virus-RNA.
The test contains an internal inhibition control (PLUS) and can be detected by all cyclers.
Using Roche LightCycler 1.5 oder 2.0:
These instruments require to perform (only once) a Color Compensation before using the new PLUS kits.
For this a SureCC Color Compensation Kit II is available.
In 1972, Norovirus was firstly detected by immuno-electron microscopic examinations of an outbreak of gastroenteritis in Norwalk (Ohio/USA) in 1968. Since 2002, this and all other genetic variations are concluded in the term Norovirus (former Norwalklike- virus). Norovirus particles have a diameter of ca. 35 nm and
consist of a capside and a single strand RNA, 7300 – 7700 nucleotides in size. Noroviruses are cause of more than 90 % of non bacterial gastroenteritis cases. One reason therefore is their stability in environment. Particularly in community facilities, e.g. hospitals, rest homes or nursery homes, Norovirus causes outbreaks of gastroenteritis that can develop to considerable scale.
Vomiting, diarrhoea and nausea are typical symptoms of a Norovirus infection. Infections can appear all year long. Nevertheless, an increased number of cases are registered during winter months (“winter vomiting disease”). Virus particles are either transferred directly from person to person, or indirectly via contaminated surfaces, devices, food (e.g. mussels, fruits or vegetables) or water. The minimal infection dose is 10-100 virus particles.
|Test Format||100 reactions|
|Sample Preparation||The use of the SureFast® PREP DNA / RNA Virus kit (F1051) is recommended.|
|Detection Limit||≤ 50 RNA copies|
|Cross Reactivity||no known interferences|
|Shelf life||24 month|