Instruction for Use HISTO TYPE Rainbow QS6
Version: 02/2021
Page 11 of 16
6.7
Import of the results file into the PlexTyper
®
software
Please follow the instructions for use for the PlexTyper
®
software for interpretation of the
data.
Open the PlexTyper
®
software. From the home screen choose
View plates with no associated
results
under
Plates
. A list opens with all tests that are not linked with raw data yet. There is a
global
Search
field at the top of the table to search the whole table.
Double click on the required test to be interpreted; this opens the results summary window . Select
Import File
and select the excel file with the correct PT RUN ID prefix (e.g. PT999.xls) exported
from the
QuantStudio™ 6 Flex System.
Initial loading of the kit file data takes approximately 45 seconds, unless it is already open, the
bottom left corner of the screen shows a progress bar. After that the results for user review are
presented.
A negative control (NTC) is used as contamination control. If DNA or contaminating amplicon is
inadvertently added to the NTC reaction a positive signal will occur. If the Cq is less than 36 it will be
detected as possible contamination by the PlexTyper
®
software and a warning message is
generated. Amplification signals above Cq 36 in the NTC are regarded as PCR artefacts and are
disregarded. If PCR contamination is suspected, it is advisable to follow local decontamination
guidelines and to exchange the reagents.
The raw data collected from the cycler-specific software will be imported into the PlexTyper
®
software. Based on the Cq values, RFUs (relative fluorescence units), quality scores and the curve
progression the PlexTyper
®
software determines the molecular genetic HLA pattern of the
specimens used (see instructions for use for PlexTyper
®
for details). Positive reactions are
determined based on the Cq ratio between the the Cq of the internal amplfication control (IAC) and
the Cq of the HLA allele specific reaction (exception see below).
Special consideration for analysis of HISTO TYPE Rainbow QS6 results:
In these kits a strong allele specific O560 signal may affect the appearance of the TAMRA IAC
amplification due to emission wavelength interference. This can result in sporadic IAC failure due
either to a false late IAC Cq value, or the reaction failing due to the fluorescence values being
negative in the last cycle. Figures below illustrate the possible appearances of the TAMRA IAC
signals when the O560 signal is strong.
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