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RespiFinder RG Panel RUO

For detection and differentiation of 16 RNA viruses, 2 DNA viruses, and 4 bacteria within 4.5 hours
  • Analysis of multiple targets in one reaction using SmartFinder technology 
  • Detection and differentiation of 22 pathogen nucleic acids in 4.5 hours 
  • High reliability using the internal control
The RespiFinder RG Panel is a qualitative multiplex PCR test to detect and differentiate 16 RNA viruses, 2 DNA viruses, and 4 bacteria within 4.5 hours. (The RespiFinder RG Panel detects 16 RNA viruses but does not differentiate rhinovirus from enterovirus.)
For Research Use Only. Not for use in diagnostics procedures. No claim or representation is intended to provide information for the diagnosis, prevention, or treatment of a disease.
Principle
The RespiFinder RG Panel is based on the SmartFinder technology, a technology that allows a highly complex analysis of multiple targets in real-time PCR. The RespiFinder RG Panel contains 23 different MultiFinder probe sets combined with 13 fluorescent labeled SMART (single-tube multiplex amplification in real time) probes that enable the detection of 22 different pathogens, plus an internal control (see table "Pathogens and corresponding target genes of the RespiFinder RG Panel").
Pathogens and corresponding target genes of the RespiFinder RG Panel
Target Gene
Control
Internal control (IC) MS2 phage lysis protein gene
 Viruses  
 Adenovirus  Hexon gene (H)
 Bocavirus  Noncapsid gene (NP1)
 Corona NL63  Nucleocapsid protein gene (NP)
 Corona OC43  Nucleocapsid protein gene (NP)
 Corona 229E  Nucleocapsid protein gene (NP)
 Corona HKU1  Nucleocapsid phosphoprotein gene (N)
 hMPV  Nucleocapsid protein gene (NP)
 Influenza A  Matrix protein gene (M1)
 Influenza B  Matrix protein gene (M1)
 Influenza A H1N1v  Neuraminidase gene
 Parainfluenza 1  Hemagglutinin-neuraminidase gene (HN)
 Parainfluenza 2  Hemagglutinin-neuraminidase gene (HN)
 Parainfluenza 3  Hemagglutinin-neuraminidase gene (HN)
 Parainfluenza 4  Major nucleocapsid protein gene (N)
 Rhinovirus/Enterovirus  5' untranslated region polyprotein gene (PP)
 RSV-A  Major nucleocapsid protein gene (N)
 RSV-B  Major nucleocapsid protein gene (N)
 Bacteria  
 Bordetella pertussis  Insertion sequence 481 (IS481)
 Chlamydophila pneumoniae  Major outer membrane gene (OmpA)
 Legionella pneumophila  Macrophage inhibitor potentiator gene (Mip)
 Mycoplasma pneumoniae  Cytadhesin protein gene (P1)
The reaction starts with a pre-amplification that combines a reverse transcription step with a PCR step to amplify the target DNA/cDNA. Subsequently, a part of the pre-amplification reaction is transferred to 2 PCR tubes. Two separate SmartFinder reactions are performed, comprising a probe hybridization step and a probe ligation and amplification step. The detection is performed using melting curve analysis.
The RespiFinder RG Panel uses the Rotor-Gene Q instrument for the detection of the pathogen(s) nucleic acids.
An internal control (IC) 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.
Procedure
The input sample is total nucleic acids extracted and purified from nasopharyngeal swabs using the QIAamp MinElute Virus Spin Kit.
The panel contains primers, probes, enzymes, and reagents for detection and differentiation of pathogen nucleic acids. Pre-amplification and hybridization are carried out on a standard block-based thermal cycler, followed by ligation and PCR on the Rotor-Gene Q instrument. Melting curve analysis on the Rotor-Gene Q enables differentiation of the pathogen nucleic acid targets.
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