Instruction for Use HISTO TYPE Rainbow QS6
Version: 02/2021
Page 14 of 16
8.1.3 Analytical specificity / cross reactive substances
Eight substances that might interfere with the assay were tested and the following concentrations
were shown to have no detrimetal effect on the results:
Substance
Maximal non-inhibitory concentration
Protein (BSA)
0.2 mg/ml
TE (Tris/EDTA, pH 8.0)
7 mM Tris, 0.7 mM EDTA
NaCl
20 mM
Ethanol
1%
Haemoglobin
0.01 mg/ml
Sodium Citrate
7 mM
DNA extraction buffer 1 (Qiagen QIAamp DNA Blood Kits)
1%
DNA extraction buffer 2 (Qiagen QIAamp DNA Blood Kits)
2%
9.
LIMITS OF THE METHOD
During DNA isolation, special attention must be paid to the fact that the RT-PCR method reacts very
sensitively to cross-contaminations. Special care should be taken to avoid contamination of kit
reagents and other laboratory materials with amplicons or DNA.
The performance of a negative control without DNA in well H12 is strongly recommended. No
fluorescence signal below 36 Cq should be detected in the NTC (H12) with molecular grade water.
In the case of signal development in the negative control, the PCR laboratory workplace may have
to be decontaminated from DNA and the reagents exchanged if necessary.
All devices (e.g. pipettes, real-
time cyclers) must be calibrated according to the manufacturer’s
specifications.
10.
INTERNAL QUALITY CONTROL
Internal quality controls for new lots can be performed with a combination of DNA specimens with
known HLA type. An internal control to verify successful amplification is included in the dried
oligomixes.
Performance of negative controls (well H12) to detect possible contaminations is recommended. For
this purpose, prepare a test without DNA (NTC), see section 6.4 Amplification.
