Background
Methods
Clinical specimen
Virus controls
Virus | Dynamic range | Efficiency | Interassay- variabilty | R2 |
---|---|---|---|---|
INF-A H1 | 1.00E + 05 - 1.00E + 01 | 1.89 | 22.7 ± 0.49 | 0.972 |
INF-A H3 | 1.00E + 05 - 1.00E + 01 | 1.89 | 22.7 ± 0.49 | 0.972 |
RSV-A | 1.00E + 06 - 1.00E + 01 | 1.98 | 22.4 ± 0.36 | 0.998 |
PIV-1 | 1.00E + 06 - 1.00E + 01 | 1.91 | 29.9 ± 0.84 | 0.993 |
PIV-2 | 1.00E + 05 - 1.00E + 01 | 1.95 | 29.3 ± 0.33 | 0.997 |
PIV-3 | 1.00E + 06 - 1.00E + 01 | 1.88 | 36.4 ± 0.30 | 0.989 |
HRV | 1.00E + 06 - 1.00E + 01 | 1.90 | 21.3 ± 0.34 | 0.991 |
AdV | 1.00E + 06 - 1.00E + 01 | 1.88 | 19.9 ± 0.17 | 0.989 |
hMPV | 1.00E + 06 - 1.00E + 01 | 1.87 | 26.7 ± 0.66 | 0.986 |
Nucleic acid extraction
xTAG Respiratory Virus Panel Fast Assay (RVP)
RespiFinder-19 and RespiFinder-SMART-22 assay
Monoplex real-time PCR
Results
Comparison of xTAG Respiratory Virus Panel Fast assay (RVP) with RespiFinder-19 assay and RespiFinder-SMART-22 assay using virus control material
Virus | Concentration [copies/ml] | RVP | RespiFinder-19 | RespiFinder- SMART-22 | Monoplex real-time PCR | ||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
und. | 1:10 | 1:100 | 1:1000 | und. | 1:10 | 1:100 | 1:1000 | und. | 1:10 | 1:100 | 1:1000 | und. | 1:10 | 1:100 | 1:1000 | ||
INF-A H1 | 1.78E+04 |
+
|
+
|
+
|
-
|
-
|
-
|
-
|
-
|
-
|
-
|
-
|
-
| 32.9 | 33.8 | 35.8 | 37.9 |
INF-A H3 | 1.42E+04 |
+
|
+
|
+
|
-
|
-
|
-
|
-
|
-
|
-
|
-
|
-
|
-
| ||||
INF-B | n. q. |
-
|
-
|
-
|
-
|
-
|
-
|
-
|
-
|
-
|
-
|
-
|
-
| x | x | x | x |
RSV-A | 3.94E+04 |
+
|
+
|
+
|
-
|
-
|
-
|
-
|
-
|
-
|
-
|
-
|
-
| 28.7 | 30.4 | 32.5 | 34.6 |
RSV-B | n. q. |
+
|
+
|
+
|
-
|
-
|
-
|
-
|
-
|
-
|
-
|
-
|
-
| x | x | x | x |
PIV-1 | 1.61E+04 |
-
|
-
|
-
|
-
|
-
|
-
|
-
|
-
|
-
|
-
|
-
|
-
| 32.8 | 33.1 | 34.9 | 36.6 |
PIV-2 | 9.19E+03 |
-
|
-
|
-
|
-
|
+
|
-
|
-
|
-
|
+
|
+
|
+
|
-
| 29.5 | 31.4 | 33.7 | 36.4 |
PIV-3 | 3.84E+04 |
-
|
-
|
-
|
-
|
-
|
-
|
-
|
-
|
-
|
-
|
-
|
-
| 34.9 | 36.8 | 38.3 | 39.8 |
CoV OC43 | n. q. |
+
|
+
|
+
|
+
|
+
|
+
|
+
|
+
|
+
|
+
|
+
|
+
| 27.6 | 28.9 | 30.1 | 31.9 |
CoV 229E | n. q. |
-
|
-
|
-
|
-
|
+
|
+
|
+
|
+
|
+
|
+
|
+
|
+
| 26.8 | 27.9 | 29.1 | 30.5 |
HRV | 2.83E+04 |
+
|
+
|
+
|
+
|
+
|
+
|
-
|
-
|
+
|
+
|
+
|
-
| 31.2 | 33.8 | 36.0 | 38.4 |
AdV | 3.56E+04 |
-
|
-
|
-
|
-
|
+
|
+
|
+
|
+
|
+
|
+
|
+
|
-
| 35.9 | 37.6 | 38.8 | 39.9 |
hMPV | 1.78E+04 |
+
|
+
|
+
|
+
|
-
|
-
|
-
|
-
|
-
|
-
|
-
|
-
| 30.2 | 32.0 | 34.8 | 38.4 |
RVP | RespiFinder-19 | RespiFinder-SMART-22 | Monoplex real-time PCR (CT) | |||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|
und. | 1:10 | 1:100 | und. | 1:10 | 1:100 | und. | 1:10 | 1:100 | und. | 1:10 | 1:100 | |
Panel 1 | 55 μl elution | 55 μl elution | 55 μl elution | 55 μl elution | ||||||||
(INF-A H1/ RSV-A/ HRV/ AdV) |
+ / + / + / +
|
+ / + / + / -
|
+ / - / + / -
|
- / + / + / +
|
- / - / + / -
|
- / - / - / -
|
- / + / + / +
|
- / - / + / -
|
- / - / + / -
| 21.8/22.3/ 20.4/28.7 | 25.8/26.9/ 26.8/30.5 | 29.6/28.3/ 30.8/31.7 |
100 μl elution | 100 μl elution | 100 μl elution | ||||||||||
+ / + / + / +
|
+ / + / + / -
|
+ / - / + / -
|
+ / - / + / +
|
+ / - / + / -
|
- / - / - / -
|
- / - / + / +
|
- / - / + / +
|
- / - / + / +
| ||||
Panel 2* | 55 μl elution | 55 μl elution | 55 μl elution | 55 μl elution | ||||||||
(INF-A H1/ INF-A H3/ RSV-A/ PIV-2) |
+ / + / + / +
|
+ / + / + / -
|
+ / + / - / -
|
- / - / + / +
|
- / - / + / +
|
- / - / + / +
|
- / - / + / +
|
- / - / + / +
|
- / - / + / +
| 24.3/24.3/ 22.8/20.4 | 28.7/28.7/ 25.5/26.4 | 31.1/31.1/ 28.7/30.4 |
100 μl elution | 100 μl elution | 100 μl elution | ||||||||||
+ / + / + / +
|
+ / + / + / -
|
+ / + / - / -
|
+ / + / - / +
|
+ / + / - / +
|
+ / + / - / -
|
+ / + / + / +
|
+ / + / + / +
|
+ / + / - / +
| ||||
Panel 3* | 55 μl elution | 55 μl elution | 55 μl elution | 55 μl elution | ||||||||
(CoV OC43/ CoV 229E/ AdV/ HRV) |
+ / - / + / +
|
+ / - / + / +
|
- / - / - / +
|
+ / + / + / +
|
+ / + / + / +
|
+ / + / + / -
|
+ / + / + / +
|
+ / + / + / +
|
+ / + / + / +
| 25.0/22.3/ 29.7/22.4 | 28.7/25.3/ 31.8/25.4 | 32.0/26.5/ 32.6/28.5 |
100 μl elution | 100 μl elution | 100 μl elution | ||||||||||
+ / - / + / +
|
+ / - / + / +
|
- / - / - / +
|
+ / + / + / +
|
+ / + / + / +
|
+ / + / + / -
|
+ / + / + / +
|
+ / + / + / +
|
+ / + / + / +
|
Detection of respiratory viruses in clinical samples
Patient | Sex | Age (yr) | RVP | RespiFinder-19 | real-time PCR |
---|---|---|---|---|---|
winter
| |||||
triple infections
| |||||
1 | m | 71 | - | INF-B, PIV-2 | INF-B, PIV-2, CMV |
double infections
| |||||
2 | m | 52 | RSV | RSV-A, CoV 229E | RSV-A, CoV 229E |
3 | m | 39 | CoV 229E | CoV 229E | CoV 229E, CMV |
4 | f | 84 | CoV HKU1 | CoV 229E, AdV | CoV HKU1, AdV |
5 | m | 59 | - | PIV-2 | PIV-2, CMV |
6 | f | 84 | x | AdV | AdV, CMV |
7 | m | 56 | - | PIV-2 | PIV-2, CMV |
8 | m | 61 | - | PIV-2, AdV | PIV-2, AdV |
9 | f | 23 | - | PIV-2, AdV | PIV-2, AdV |
10 | f | 84 | - | AdV | AdV, CMV |
11 | f | 83 | - | PIV-2, RSV-A | PIV-2, RSV-A |
12 | m | 85 | - | PIV-2, AdV | PIV-2, AdV |
13 | m | 79 | - | AdV, PIV-3 | AdV, PIV-3 |
14 | f | 76 | - | PIV-2, AdV | PIV-2, AdV |
15 | m | 56 | - | AdV, PIV-3 | AdV, PIV-3 |
16 | m | 71 | x | AdV | AdV, CMV |
17 | m | 74 | - | INF-A, PIV-2 | INF-A, PIV-2 |
mono infections
| |||||
18 | f | 84 | AdV | AdV | AdV |
19 | f | 80 | hMPV | hMPV | hMPV |
20 | m | 65 | PIV-3 | PIV-3 | PIV-3 |
21 | m | 73 | - | AdV | AdV |
22 | m | 70 | - | AdV | AdV |
23 | m | 79 | - | PIV-2 | PIV-2 |
24 | f | 62 | - | AdV | AdV |
25 | f | 72 | HRV | HRV | HRV |
26 | f | 82 | HRV | HRV | HRV |
27 | m | 80 | - | AdV | AdV |
28 | m | 71 | CoV 229E | CoV 229E | n. d. |
29 | m | 64 | - | AdV | AdV |
30 | m | 58 | - | AdV | AdV |
31 | f | 46 | HRV | - | HRV |
32 | f | 75 | HRV | HRV | HRV |
33 | m | 52 | - | INF-B | INF-B |
summer
| |||||
double infections
| |||||
34 | f | 74 | HRV | HRV | HRV, CMV |
35 | m | 70 | x | AdV | AdV, CMV |
mono infections
| |||||
36 | m | 59 | HRV | - | HRV |
37 | f | 77 | - | - | CMV |
38 | f | 78 | - | - | CMV |
39 | m | 81 | - | - | CMV |
40 | f | 43 | - | HRV | HRV |
41 | m | 73 | - | AdV | AdV |
42 | f | 91 | HRV | HRV | HRV |
43 | m | 66 | - | PIV-2 | PIV-2 |
44 | m | 59 | - | AdV | AdV |
45 | f | 65 | - | AdV | AdV |
46 | f | 48 | - |
L. pneumophila
|
L. pneumophila
|
47 | m | 66 | - | AdV | AdV |
Discussion
RVP | RespiFinder-19 | RespiFinder-SMART-22 | |
---|---|---|---|
Manufacturer
| Abbott Molecular | PathoFinder | PathoFinder |
CE IVD/FDA
| yes/yes | yes/no | yes/no |
TEST SPECIFICATIONS
| |||
Pathogens
| influenza A + B | influenza A + B | influenza A + B |
RSV-A + B | RSV-A + B | RSV-A + B | |
adenovirus | adenovirus | adenovirus | |
hMPV | hMPV | hMPV | |
PIV-1, -2, -3, -4 | PIV-1, -2, -3, -4 | PIV-1, -2, -3, -4 | |
CoV 229E, NL63, OC43, HKU1 | CoV 229E, NL63, OC43 | CoV 229E, NL63, OC43, HKU1 | |
HRV | HRV | HRV | |
enterovirus | - | enterovirus | |
bocavirus | - | bocavirus | |
- |
B. pertussis
|
B. pertussis
| |
- |
C. pneumoniae
|
C. pneumoniae
| |
-- |
L. pneumophilaM. pneumoniae
|
L. pneumophilaM. pneumoniae
| |
Sample volume
| 500 μL | 200 μL | 200 μL |
Elution volume
| 55 μL | 100 μL | 100 μL |
PCR volume
| 10 μL | 10 μL | 10 μL |
Principle of detection
| fluorescence bead array | capillary electrophoresis | melt curve analysis |
PERFORMANCE
| |||
Hands-on-time
| 0.80 h | 1.75 h | 1.25 h |
Time-to-result
| 3.5 h | 7.5 h | 6.0 h |
COSTS
| |||
Costs reagents *
,1
| 90-100 € | 50-60 € | 50-60 € |
Costs equipment *
| 57-69,000 € | 100-120,000 € | 33-40,000 € |
ACCOMPLISHMENT
| |||
Handling
| + | + | ++ |
Personal training duration/qualification
| 2 d/very low | 2 d/very low2
| 2 d/low |
Technical equipment
| extraction system, thermocycler, Luminex 200 | extraction system, thermocycler, DNA-Sequencer ABI3500 | extraction system, thermocycler, Rotorgene Q |