RespiFinder® RG Panel
Performance Characteristics
REF
RespiFinder RG Panel, Version 1,
Check
4692163
availability
of
new
electronic
labeling
revisions
at
www.qiagen.com/p/RespiFinder-RG-Panel-CE before test execution. The
current revision status is indicated by the issue date (format: month/year).
Limit of detection (LOD)
The limit of detection (LOD) in consideration of the purification (sensitivity limit) was assessed for the
RespiFinder RG Panel using whole virus and bacterial specimens in Universal Transport Medium
(UTM) in combination with the QIAamp® MinElute® Virus Spin Kit extraction.
The limit of detection in consideration of the purification of the RespiFinder RG Panel was
determined using a dilution series of viral and bacterial strains (ATCC®, Zeptometrix®, Viral Culture)
in UTM. These were subjected to RNA/DNA extraction using the QIAamp MinElute Virus Spin Kit
(extraction volume: 200 μl, elution volume: 60 μl). Each of the 5 dilutions was analyzed with the
RespiFinder RG Panel with 11 replicates each. The results were determined by hit rate (≥95% hit
rate was defined as the analytical detection limit for the respective pathogen). An overview of the
limit of detection for all pathogens tested is shown in Table 1.
Table 1. Limit of detection in consideration of the purification (QIAamp MinElute Virus Spin
Kit) of the RespiFinder RG Panel
Subtype/
Pathogen
strain
Hit rate
Source
LOD titer
(%)
Viruses
Adenovirus
B3
Bocavirus
n.a.†
Coronavirus
229E
ATCC*
Recombinant
plasmid
ATCC
* American Type Culture Collection, USA.
†
10(0.40) TCID50/0.2 ml
95
4 x 10–3 fg/μl
100
10(–3.95) TCID50/0.2 ml
100
Table continued on next page
n.a.: not applicable.
November 2013
Sample & Assay Technologies
Table 1. Continued
Subtype/
Pathogen
strain
Hit rate
Source
LOD titer
(%)
Viruses (continued)
Coronavirus
OC43
ATCC*
10(–2.25) TCID50/0.2 ml
100
Coronavirus
NL63
Zeptometrix
n.a.†
100
Coxsackievirus
A9
ATCC
10(–0.60) TCID50/0.2 ml
95
hMPV
B3
Zeptometrix
n.a.
100
ATCC
10(–1.50) TCID50/0.2 ml
100
ATCC
10(0.10) TCID50/0.2 ml
100
Influenza A
Influenza A
H3N2
Victoria
H1N1
Virginia
Influenza B
Maryland
ATCC
10(–1.17) TCID50/0.2 ml
100
Parainfluenza
1
ATCC
10(–3.25) TCID50/0.2 ml
95
Parainfluenza
2
ATCC
10(–1.25) TCID50/0.2 ml
100
Parainfluenza
3
ATCC
10(–0.25) TCID50/0.2 ml
100
Parainfluenza
4a
ATCC
10(–0.75) TCID50/0.2 ml
100
Rhinovirus
16
ATCC
10(1.50) TCID50/0.2 ml
100
RSV-A
2
ATCC
10(–2.25) TCID50/0.2 ml
95
RSV-B
18537
ATCC
10(–1.75) TCID50/0.2 ml
100
B. pertussis
n.a.
ATCC
8 x 10–4
100
L. pneumophila
n.a.
ATCC
10(–1.70) bact/0.2 ml
100
M. pneumoniae
n.a.
ATCC
10–2
100
Bacteria
* American Type Culture Collection, USA.
†
n.a.: not applicable.
Performance Characteristics: RespiFinder RG Panel
page 2 of 8
Specificity
The specificity of the RespiFinder RG Panel is first and foremost ensured by the selection of the
primers and probes, as well as the selection of stringent reaction conditions. The primers and
probes were checked for possible homologies to all sequences published in gene banks by
sequence comparison analysis. The detectability of all relevant genotypes has thus been ensured by
RespiFinder RG Panel runs on a Rotor-Gene® Q MDx instrument with the following genotypes (see
Table 2).
Table 2. Testing of the specificity of relevant strains
Internal control
Pathogen
Source
FAM™/ROX™
FAM/Cy 5
(FAM/BHQ®))
Adenovirus B7
ATCC*
+
–
+
Adenovirus B11
ATCC
+
–
+
Adenovirus B14
ATCC
+
–
+
Adenovirus C1
ATCC
+
–
+
Adenovirus C5
ATCC
+
–
+
Adenovirus C6
ATCC
+
–
+
Adenovirus E4
ATCC
+
–
+
Coxsackievirus A10
ATCC
–
+
+
Coxsackievirus A24
ATCC
–
+
+
Coxsackievirus B1
ATCC
–
+
+
Coxsackievirus B2
ATCC
–
+
+
Coxsackievirus B3
ATCC
–
+
+
Coxsackievirus B5
ATCC
–
+
+
Echovirus 4
ATCC
–
+
+
Echovirus 11
ATCC
–
+
+
Echovirus 30
ATCC
–
+
+
ATCC
+
–
+
Influenza A H1N1 New
Jersey
* American Type Culture Collection, USA.
Performance Characteristics: RespiFinder RG Panel
®
Table continued on next page
page 3 of 8
Table 2. Continued
Internal control
Pathogen
Source
FAM/ROX
FAM/Cy5
(FAM/BHQ))
Parainfluenza 4b
ATCC*
+
–
+
Rhinovirus 1A
ATCC
–
+
+
Rhinovirus 1B
ATCC
–
+
+
Rhinovirus 2
ATCC
–
+
+
Rhinovirus 30
ATCC
–
+
+
Rhinovirus 39
ATCC
–
+
+
Rhinovirus 60
ATCC
–
+
+
RSV-A long
ATCC
+
–
+
RSV-B WV
ATCC
+
–
+
* American Type Culture Collection, USA.
A potential cross-reactivity of the RespiFinder RG Panel was tested using the control group listed in
Table 3. None of the tested pathogens were reactive.
Table 3. Testing the specificity of the kit with potentially cross-reactive pathogens
Internal control
Pathogen
FAM/ROX
FAM/Cy5
(FAM/BHQ)
Cytomegalovirus
–
–
+
Epstein-Barr virus
–
–
+
Herpes simplex virus
–
–
+
Varicella-zoster virus
–
–
+
A. baumannii
–
–
+
A. fumigatus
–
–
+
Viruses
Bacteria
Table continued on next page
Performance Characteristics: RespiFinder RG Panel
page 4 of 8
Table 3. Continued
Internal control
Pathogen
FAM/ROX
FAM/Cy5
(FAM/BHQ)
C. albicans
–
–
+
C. krusei
–
–
+
C. lusitaniae
–
–
+
E. cloacae
–
–
+
E. coli
–
–
+
H. influenzea
–
–
+
K. pneumoniae
–
–
+
K. oxytoca
–
–
+
M. catarrhalis
–
–
+
M. tuberculosis
–
–
+
P. aeruginosa
–
–
+
S. aureus
–
–
+
S. marcescens
–
–
+
S. pneumoniae
–
–
+
Bacteria (continued)
Co-infections
The RespiFinder RG Panel was validated for multiple infections to confirm detection of relevant coinfections. As shown in Table 4, co-infections of influenza A, influenza B, RSV-A, or RSV-B with
adenovirus or rhinovirus were tested at different concentrations of both pathogens. All co-infections
were detected.
Performance Characteristics: RespiFinder RG Panel
page 5 of 8
Table 4. Testing the specificity of the kit with multiplex infections
Internal control
Pathogen 1
Pathogen 2
FAM/ROX
FAM/Cy5
(FAM/BHQ))
+
–
+
+
–
+
+
+
+
+
+
+
+
–
+
+
–
+
+
+
+
+
+
+
+
–
+
+
–
+
+
+
+
+
+
+
Adenovirus B3
— 10x LOD
Adenovirus B3
Influenza A H3N2
— 100x LOD
Victoria — 10x LOD
Rhinovirus 16
— 10x LOD
Rhinovirus 16
— 100x LOD
Adenovirus B3
— 10x LOD
Adenovirus B3
Influenza B Maryland
— 100x LOD
— 10x LOD
Rhinovirus 16
— 10x LOD
Rhinovirus 16
— 100x LOD
Adenovirus B3
— 10x LOD
Adenovirus B3
— 100x LOD
RSV-A 2 — 10x LOD
Rhinovirus 16
— 10x LOD
Rhinovirus 16
— 100x LOD
Table continued on next page
Performance Characteristics: RespiFinder RG Panel
page 6 of 8
Table 4. Continued
Internal control
Pathogen 1
Pathogen 2
FAM/ROX
FAM/Cy5
(FAM/BHQ))
+
–
+
+
–
+
+
+
+
+
+
+
Adenovirus B3
— 10x LOD
Adenovirus B3
RSV-B 18537 —
— 100x LOD
10x LOD
Rhinovirus 16
— 10x LOD
Rhinovirus 16
— 100x LOD
Robustness
The verification of the robustness allows the determination of the total failure rate of the RespiFinder
RG Panel internal control. To verify the robustness, 30 negative nasopharyngeal swab samples in
Universal Transport Medium (UTM) were tested with the RespiFinder RG Panel. After extraction
using the QIAamp MinElute Virus Spin Kit (extraction volume: 200 μl, elution volume: 60 μl), these
samples were analyzed with the RespiFinder RG Panel. In all samples the internal control was
detected and valid. Thus, the robustness of the RespiFinder RG Panel is ≥99%.
Interfering substances
The RespiFinder RG Panel was tested for the interference by nasal spray (Otrivin®, Novartis®) and
human genomic DNA. All pathogens listed in Table 1 (pages 1–2) were tested at a concentration of
3x LOD in the presence of 50 ng human DNA. Additional negative control samples were tested in
the presence of human DNA. The correct pathogens were detected in all samples containing
additional human DNA, and the internal control was detected in the negative control samples,
showing no interference of human DNA on the performance of RespiFinder RG Panel.
Ten representative pathogens from Table 1 (adenovirus B3, coronavirus OC43, hMPV B3, influenza
A H1N1 Virginia, influenza B Maryland, parainfluenza 1, rhinovirus 16, RSV-A 2, L. pneumophila,
and M. pneumoniae) were tested at a concentration of 3x LOD in the presence of 10% (v/v) nasal
spray. Additional negative control samples were tested in the presence of 10% nasal spray. The
correct pathogens were detected in all samples containing nasal spray, and the internal control was
detected in the negative control samples, showing no interference of nasal spray (10% of sample:
swab in Universal Transport Medium) on the performance of RespiFinder RG Panel.
Performance Characteristics: RespiFinder RG Panel
page 7 of 8
For up-to-date licensing information and product-specific disclaimers, see the respective QIAGEN
kit handbook or user manual. QIAGEN kit handbooks and user manuals are available at
www.qiagen.com or can be requested from QIAGEN Technical Services or your local distributor.
Trademarks: QIAGEN®, QIAamp®, MinElute®, Rotor-Gene® (QIAGEN Group); ATCC® (American Type Culture Collection);
BHQ® (Biosearch Technologies, Inc.); Cy® (GE Healthcare); FAM™, ROX™ (Life Technologies); Novartis®, Otrivin® (Novartis);
RespiFinder® (PathoFinder BV); Zeptometrix® (Zeptometrix Corp.).
Nov-13 © 2013 QIAGEN, all rights reserved.
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