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Thermo Scentific

Remel - Diagnostic Tests

Sector: Clinical


Code: R240396

The ProSpecT Rotavirus test is a qualitative enzyme immunoassay for the detection of rotavirus (Group A) in human faecal samples as an aid in the diagnosis of acute gastroenteritis. The test utilises a polyclonal antibody to detect group specific proteins, including the major inner capsid protein (VP6), present in Group A rotaviruses.

Human serotypes of Group A rotavirus are a major cause of gastroenteritis in young children throughout the world. Rotavirus also causes gastroenteritis in older children and elderly populations. The virus is commonly associated with nosocomial infections in paediatric wards and neonatal nurseries.

The laboratory diagnosis of rotavirus infections plays an important role in patient management and enables effective management and control of outbreaks. Laboratory diagnosis relies on direct detection of the virus or viral antigens in faecal specimens.

Enzyme immunoassays using specific monoclonal or polyclonal antibodies offer a rapid, sensitive and specific method for the detection of rotavirus in faecal specimens.

Principle of the Test
The ProSpecT Rotavirus test utilises a polyclonal antibody in a solid-phase sandwich enzyme immunoassay to detect group specific antigen present in Group A rotaviruses. Breakapart microwells are coated with a rotavirus specific polyclonal antibody. Faecal suspension or control sample is added to the microwell and incubated simultaneously with a rotavirus specific polyclonal antibody conjugated to horseradish peroxidase. Rotavirus antigen present in the sample is captured between antibody on the solid phase and the enzyme conjugated antibody. After 60 minutes incubation at room temperature, the microwells are washed with working strength wash buffer to remove excess specimen and any unbound enzyme labelled antibody. A chromogen is added to the microwells and incubated for 10 minutes at room temperature. The presence of specifically bound enzyme labelled antibody in the microwells results in a colour change which is stopped by the addition of acid. Colour intensity significantly above background levels is indicative of the presence of rotavirus antigen in the specimen or control.

Fact Sheet

Tests per kit 96
Shelf life  18 months (from date of manufacture) 
Incubations2 (60 minutes/10 minutes both at room temperature)
Time to result <2 hours
ControlsPositive and negative controls included in the kit 
ReadingSpectophotometric (450nm) or visual determination 

Kit Components

Microtitration plate: a 96-well plate of twelve, 8-microwell break-apart strips coated with a rotavirus specific rabbit polyclonal antibody
Sample Diluent (120ml): tris buffered saline containing antimicrobial agent and red dye
Positive Control (4ml): inactivated bovine rotavirus in buffer containing antimicrobial agent
Negative Control (4ml): tris buffered saline containing antimicrobial agent and red dye
Conjugate (12ml): Rotavirus specific rabbit polyclonal antibody conjugated to horseradish peroxidase in a buffered protein solution containing antimicrobial agent and blue dye
Substrate TMB (12ml): 3,3’-5,5’-tetramethylbenzidine in a mildly acidic buffer
Stop Solution (12ml): 0.46mol/l sulphuric acid
Wash Buffer (120ml): concentrated x 10. Phosphate buffered solution containing antimicrobial agent and detergent.

Materials Required but not Provided

Faecal specimen collection containers
Clean, screw capped disposable containers for preparation of faecal specimen
Clean, absorbent paper (onto which microwells can be tapped dry)
Precision micropipettes and disposable tips to deliver 50µl, 100µl and 1,000µl
Waste discard container with suitable fresh disinfectant
Wash bottle for wash buffer
Distilled or deionised water

The test is intended for in vitro diagnostic use. Anyone performing an assay with this product must be trained in its use and must be experienced in laboratory procedures.
The wash buffer contains potential skin sensitiser (<1%v/v). Avoid skin contact. Wear disposable vinyl or nitrile gloves.

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