Features and Benefits

  • Detects all members of the Mycobacterium tuberculosis
              Complex
  • Detects all clinically significant nontuberculous mycobacteria
  • Diagnosis within 75 minutes
  • Internal Control included
  • Positive Control included
  • Generic RealAccurate® Quadruplex amplification protocol

Overview

Tuberculosis (TB), which is caused by members of the Mycobacterium tuberculosis complex (MTBC: M. tuberculosis, M. bovis, M. microti, M. africanum and M. canetti), remains a global health problem. Nontuberculous mycobacteria (NTM) are a diverse group of bacteria that include more then 200 different species, some of which can infect humans and cause pulmonary disease that resembles TB. A rapid diagnosis of MTBC and NTM is particulary important since conventional culturing takes 2-8 weeks depending on the Mycobacterium species. The differentiation of MTBC types from NTM types is of primary importance for infection control and choice of antimicrobial therapy.

The RealAccurate® Quadruplex Mycobacteria PCR Kit can detect and differentiate MTBC and NTM DNA. The Quadruplex Mycobacteria assay is a PCR assay which is designed for fast and sensitivive detection on real-time PCR instruments using 3 different detection channels (FAM, Texas Red and TYE665).

Targets

  • Members of the Mycobacterium tuberculosis complex
            (M. tuberculosis, M. bovis, M. microti, M. africanum and M. canetti)
  • Nontuberculous mycobacteria

Detection systems

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

Improvements

The RealAccurate® Quadruplex Mycobacteria PCR Kit has been improved regarding the previous RealAccurate® Mycobacterium tuberculosis PCR Kit:

  • Detection of nontuberculous mycobacteria
  • Addition of master mix
  • Addition of Internal Control which controls the complete
             sample process

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 Mycobacteria PCR kit as a control for the assay procedure.

Procedure

The RealAccurate® Quadruplex Mycobacteria PCR Kit contains a ready to use optimized mix of primers and probes for the detection and differentiation of MTBC and NTM. The master mix contains Taq DNA polymerase and has been extensively validated in combination with the Mycobacteria primer/probe mix to allow for detection of low copy numbers of Mycobacterium species.

An Internal Control is included in the assay to discriminate between true negative samples and false negative samples due to nucleic acid degradation, PCR inhibition or test failure. The RealAccurate® Quadruplex Mycobacteria PCR Kit also contains a Positive Control consisting of the targets detected in the kit.

Validation

The RealAccurate® Quadruplex Mycobacteria PCR Kit is validated on LightCycler® 480 (Roche) and Rotor-Gene® Q (QIAGEN) for amplification after nucleic acid extraction using NucliSENS® easyMAG® (bioMérieux) or QIAamp® MinElute Virus Spin columns (QIAGEN) of sputum, BAL, decontaminated sediment (of BAL or sputum) and MGIT™ samples.

Internal Amplification Control

The Internal Control supplied in the  RealAccurate® Mycobacteria 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