Features and Benefits

  • Discrimination between Bordetella pertussis, Bordetella                           parapertussis and Bordetella holmesii.
  • Internal Control included
  • Positive Control included
  • Diagnosis within 75 minutes
  • Generic RealAccurate® Quadruplex amplification protocol

Overview

Bordetella pertussis continues to be a major global pathogen to affect infants, children and adults. Despite major disease reduction due to the introduction of pertussis vaccines, epidemics of pertussis persist. Although Bordetella pertussis has been classically identified as the sole agent responsible for whooping cough, other species i.e. Bordetella parapertussis and Bordetella holmesii can also cause symptoms that resemble whooping cough.

The RealAccurate® Quadruplex Bordetella PCR Kit is a multiplex PCR-based assay for the detection and differentiation of three Bordetella species that are either responsible for whooping cough or resemble whooping cough in humans: Bordetella pertussis, Bordetella parapertussis and Bordetella holmesii.

Targets

  • Bordetella pertussis
  • Bordetella parapertussis
  • Bordetella holmesii.

Detection systems

Detection is performed on real-time PCR instruments capable to detect FAM, MAX, Texas Red and TYE665.

Analysis and Interpretation

Real-time PCR using Taqman Probe detection is used for interpretation.

The Internal Control reaction controls for the quality of the used sample and controls for inhibiting factors present in the sample. A negative test result is validated by a positive Internal Control result.

A Positive Control is included in the RealAccurate® Quadruplex Bordetella PCR kit as a control for the assay procedure.

Procedure

The RealAccurate® Quadruplex Bordetella PCR Kit is a sensitive real-time PCR test for detection and differentiation of: Bordetella pertussis, Bordetella parapertussis and Bordetella holmesii. The kit contains a ready to use mix of primers and probes and a master mix comprising a hot start Taq DNA polymerase.

An Internal Control is included to distinguish false negatives due to inhibition from true negatives. Kit components have been extensively validated to ensure the detection of low copy numbers of Bordetella nucleic acid.

Validation

The RealAccurate® Quadruplex Bordetella PCR Kit is validated on LightCycler® 480 (Roche), Rotor-Gene® Q (QIAGEN), CFX96™ (Bio-Rad) and Mic qPCR Cycler (Bio Molecular Systems) for amplification after nucleic acid extraction using NucliSENS® easyMAG® (bioMérieux), MagNA Pure 96 (Roche) or QIAamp® cador® Pathogen Mini Kit (QIAGEN) of nasopharyngeal swab samples.

Internal Amplification Control

The Internal Control supplied in the RealAccurate® Bordetella PCR Kit is a M13 bacteriophage solution and is supplied as a control for lysis, DNA extraction, real-time PCR performance and to check for possible PCR inhibition

Quality

  • Designed and manufactured under EN ISO 13485:2012
  • CE-IVD marked
  • Validated on QCMD panels
  • Validated on clinical samples

References