Features and Benefits

  • 18 viral + 4 bacterial pathogens in a single assay
  • As sensitive as monoplex Real Time PCR
  • Internal Amplification Control included

Overview

Acute respiratory tract infection (RTI) is the most widespread type of acute infection in adults and children and is a significant cause of disease in immunocompromised patients. Both viruses and bacteria can cause acute RTI, and the number of causative pathogens is large and diverse.

RespiFinder® 22 is a ready to use set of primers, probes and enzymes for the simultaneous detection and differentiation of 22 respiratory pathogens, with the same sensitivity and specificity as monoplex Real Time PCR. Detection is based on fragment size analysis.

Sensitive and comprehensive diagnosis of the causative pathogen(s) can be achieved within a single working day. These fast resuts will improve therapy and will assist in the prevention of unnecessary use of antiobiotics or hospital admissions.

Targets

Influenza A

Influenza A(H1N1)pdm09

Influenza B

Parainfluenza-1

RSV-A

Parainfluenza-2

RSV-B

Parainfluenza-3

Human Metapneumovirus

Parainfluenza-4

Rhinovirus/Enterovirus

Bocavirus (type 1)

Adenovirus

Coronavirus NL63

Mycoplasma pneumoniae

Coronavirus HKU1

Chlamydophila pneumoniae

Coronavirus 229E

Legionella pneumophila

Coronavirus OC43

Bordetella pertussis

 

Detection systems

RespiFinder® 22  is validated using the 3500 Genetic Analyzer capillary elctrophoresis system of Applied Biosystems

Analysis and Interpretation

Analysis of RespiFinder® 22 products is performed using the fragment size analysis mode on a capillary electrophoresis system. Fragment sizes of the amplified products vary in length from approximately 150 to 500 basepairs. Since actual fragment sizes might slightly vary from predicted sizes, a Reference Marker, containing all fragments that can be obtained with the RespiFinder® 22 kit, is included in the kit. With the Reference Marker the actual fragment sizes on a partucular capillary electhropheresis system can be determined.

Procedure

After a gene-specific multiplex reverse transcription step, two unique MultiFinder® probes are hybridized to the (c)DNA of  the pathogen(s) present in the specimen. Hybridized probes are then joined by a ligase enzyme. Subsequently, the ligated probes are amplified using a single PCR primer pair for all targets, of which one primer is labeled with a fluorescent dye.

The detection of the amplified labelled amplicons is by size fragment analysis.

The Internal  Control (IC), which is added at the beginning of the procedure is included in the assay to validate a negative sample result.

Validation

RespiFinder® 22 is validated on QCMD panels and clinical Nasopharyngeal swabs samples.

Internal Amplification Control

The RespiFinder®  22 contains an Internal Amplification Control which is added to the sample in the nucleic acid extraction procedure. The Internal Amplification Control is a RNA transcript from the encephalomyocarditis (EMC) virus. The Internal Amplification Control is supplied as a control for the RespiFinder® 22 procedure and to check for possible PCR inhibitors present in the sample.

A negative test result is validated by the presence of the Internal Amplification Control result.

Quality

  • Designed and manufactured under EN ISO13485:2012
  • CE-IVD marked 
  • Validated on QCMD panels
  • Validated on clinical samples

References