Features and Benefits

  • 5  viral and 1 atypical bacteria multiplex respiratory pathogen
             PCR Kits
  • Positive control containing 4 targets, one for each detection
              channel per kit
  • All kits contain Internal Control
  • Master mix included
  • Generic amplification protocol for all kits.

Overview

Acute respiratory tract infection is the most widespread type of acute infection in adults and children. The number of pathogens involved is numerous. Changes in the population as it ages and the increased number of patients with immunocompromised conditions have increased the number of individuals at risk.

The RealAccurate® Quadruplex Respiratory PCR Kits are six separate multiplex PCR Kits which can detect and differentiate 22 respiratory pathogens in total. The Quadruplex assays are one-step reverse transcriptase PCR assays which are designed for fast detection on real-time PCR instruments using 4 different detection channels (FAM, MAX, Texas Red and TYE665).

Targets

PF0970-R RealAccurate® Quadruplex Influenza PCR Kit

  • Influenza A virus
  • Influenza B virus
  • Influenza A(H1N1) pdm09 virus
  • Internal Control

PF0971-R RealAccurate® Quadruplex Corona PCR Kit

  • Coronavirus 229E
  • Coronavirus OC43
  • Coronavirus NL63/HKU1
  • Internal Control

PF0972-R RealAccurate® Quadruplex Parainfluenza PCR Kit 

  • Parainfluenza virus 1
  • Parainfluenza virus 2/4
  • Parainfluenza virus 3
  • Internal Control

PF0973-R RealAccurate® Quadruplex RSV/hMPV PCR Kit

  • Respiratory Syncytial Virus A
  • Respiratory Syncytial Virus B
  • Human metapneumovirus A/B
  • Internal Control

PF0974-R RealAccurate® Quadruplex Adeno/Boca/Rhino/Entero PCR Kit

  • Adenovirus
  • Bocavirus
  • Rhinovirus/Enterovirus
  • Internal Control

PF0966-R RealAccurate® Quadruplex Atypical Bacteria PCR Kit 

  • Mycoplasma pneumoniae
  • Chlamydophila pneumononiae
  • Legionella pneumophila
  • Bordetella pertussis

Detection systems

Detection is performed on a real-time PCR instrument, capable of detection of FAM, MAX, Texas Red and TYE665.

Analysis and Interpretation

Real-time PCR using Taqman Probe detection is used to interpret the results.

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 Respiratory PCR Kits as a control for the assay procedure.

Procedure

The RealAccurate® Quadruplex Respiratory PCR Kits are composed of ready to use optimized mixtures of primers and probes for the detection of 18 viruses and 4 bacteria which can cause respiratory tract infections. The master mix¹ is composed of a mixture of reverse transcriptase and Taq DNA polymerase for quantative PCR using target specific probes and enables for detection of low copy numbers of the respiratory pathogens. As most of  the respiratory pathogens of the Quadruplex panels consist of RNA, the real-time PCR is preceded by a reverse transcription step in which the viral RNA is converted into cDNA using highly specific primers. Subsequently, the cDNA is subjected to PCR using target-specific primer/probe combinations. Both steps take place in the same tube (one-step reverse transcriptase PCR).

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

¹ Master mix of Atypical Bacteria PCR Kit only contains Taq DNA polymerase, no reverse transcriptase


 

Validation

The RealAccurate® Quadruplex Respiratory PCR Kits are validated on LightCycler® 480 (Roche) and Rotor-Gene® Q (QIAGEN) for amplification after nucleic acid extraction of nasopharyngeal swabs using NucliSENS® easyMAG® (bioMérieux).

Internal Amplification Control

The Internal Control supplied in the viral PCR Kits is a MS2 bacteriophage solution and is supplied as a control for lysis, RNA/DNA extraction, the RealAccurate® Quadruplex Respiratory PCR assay performance, and to check for possible PCR inhibition.

Quality

  • Designed and manufactured under EN ISO 13485:2012
  • CE-IVD marked (viral Quadruplex Respiratory PCR Kits)
  • Validated on QCMD panels
  • Validated on clinical samples