BÜHLMANN Quantum Blue sCAL Manual Del Usuario página 4

1. Dilution of Serum Samples
• Prior to the measurement, dilute serum samples 1:10
with chase buffer (B-LFMRP-CB) (e.g. mix 20 µL
sample with 180 µL chase buffer) in a test tube.
• Mix well (e.g. using a vortex mixer).
2. Lateral Flow Assay Procedure and Readout
• Load the lot specific parameters from the RFID chip
card.
• Add 60 µL of diluted sample onto the sample loading
port of the Test Cassette.
• Incubate for 12 minutes (set a timer manually).
• Load the Test Cassette onto the Test Cassette holder of
the reader.
• Scan the Test Cassette with the Quantum Blue
by pressing the start ("ENTER") button immediately.
• For Low / High Controls: Repeat step 2 using 60 µL of
Control instead of diluted serum.
Remark: Please refer to the Quantum Blue
manual to learn about the basic functions and how to
initialize and operate the reader, especially how to select
test methods, and how to load lot-specific parameters from
the RFID chip card in order to get the samples measured.
• If the precision of the assay does not correlate with the
established limits and repetition, exclude errors in
technique, and check the following issues: i) pipets,
thermometers and timers, ii) expiration date of reagents,
iii) storage and incubation conditions.
• Result of the self-test of the Quantum Blue
performed at startup of the instrument has to be valid.
• For a valid test result, the Control Line (C) must be
visible (see appendix I, Figures 1A and 1B). It is used
as functional test control only and cannot be used for
the interpretation of the Test Line (T). If the Test Line
(T) is not detectable after the incubation (Figure 1A), the
concentration of MRP8/14 present in the sample is
below the detection limit. If a Test Line (T) is detectable
after the incubation
concentration in the sample is calculated by the
®
Quantum Blue Reader.
• If only the Test Line (T) is detectable after the
incubation (Figure 1C), the test result is invalid and the
MRP8/14 assay has to be repeated using another Test
Cassette.
• If neither the Control Line (C) nor the Test Line (T) are
detectable after the incubation (Figure 1D), the test
result is invalid and the assay has to be repeated using
another Test Cassette.
• As the Quantum Blue
evaluation of the test (T) and control (C) lines, an
additional validity check of the Control Line (C) is
undertaken. If the signal intensity of the Control Line (C)
is below a lot-specific threshold after the incubation
Revision date: 2016-11-23
ASSAY PROCEDURE
®
QUALITY CONTROL
VALIDATION OF RESULTS
(Figure 1B), the
®
Reader allows for a quantitative
time, the test result is also invalid and the MRP8/14
assay has to be repeated using another Test Cassette.
STANDARDIZATION AND INTERPRETATION OF
• The lateral flow assay is calibrated against the
BÜHLMANN sCAL ELISA (order code: EK-MRP8/14).
• The
lot-specific standard curve to calculate the MRP8/14
concentration. The assay range is defined being
between 0.5 and 10 µg/mL.
• For quantitative measurements of samples reading
above 10 µg/mL dilute the serum samples additionally
1:10 (total 1:100) and test it again according to the
assay procedure. The measured concentration must
then be multiplied by the reciprocal volume factor to
®
Reader
obtain the final result.
• The reagents supplied with this kit are optimized to
Reader
measure human MRP8/14 in serum samples.
• MRP8/14 results can be used supplementary to other
information in order to establish the diagnosis.
The Quantum Blue® sCAL test helps quickly establish a
first estimate of the acute inflammatory status of the
patient.
The following reference values might help characterize the
inflammatory status of a patient. Patients with RA* (n = 20)
show mean levels of serum MRP8/14 of 6.25 µg/mL
(SD: 3.43 µg/mL) whereas patients responding to treat-
®
Reader
ment (n = 20)
(SD: 1.45 µg/mL) (ref 1,2).
MRP8/14 normal values were determined using serum
samples from apparently healthy and asymptomatic blood
donors (adult men and woman at the age 18-70 years). 122
serum samples were analysed by EK-MRP/8/14 due to the
good correlation (R
MRP25.
The MRP8/14 levels of patients with RA were significantly
higher compared to healthy controls (n = 122) with a
median serum MRP8/14 concentration of 1.16 µg/mL and
th
the 95 percentile at a concentration of 2.92 µg/mL.
MRP8/14
*RA (Rheumatoid arthritis)
Limit of Blank (LoB): 0.25 µg/mL MRP8/14
Limit of Detection (LoD): 0.42 µg/mL MRP8/14
Repeatability:
Quantum Blue
samples within 20 days. Each sample was tested in
duplicates twice a day according to the assay procedure.
The repeatability varied between 11.9 and 25.4 %.
4/24
®
Quantum Blue
RESULTS & INTERPRETATION
showed mean levels
2 = 0.94, slope = 0.93) with the LF-
PERFORMANCE CHARACTERISTICS
17.2 % CV. The
®
sCAL assay was calculated from 10
RESULTS
Reader uses a
LIMITATIONS
of 2.92 µg/mL
repeatability of the
LF-MRP25