Chroni
Biomedical
Page 4 of 10
GLT are, in part, derived from what is
referred to as the "repetitive" sequences
that flank the majority of all genes
including the prion gene, the source of
PrP
res
. Nucleic acids are extracted from
serum or plasma and amplified. The
signal strength of the test sample is
compared to healthy and BSE control
samples. Samples that are more than
five standard deviations (detection limit)
above the normal healthy controls and
non-template controls are considered
reactive. In accordance with GLP
procedures, reactive samples are rerun in
duplicate to confirm that they are
repeatedly reactive.
Technical Details
Serum Collection
1) Collect blood from the tail artery or
other suitable vein into a pre-labeled
serum collection tube or container.
2) Store the blood at 2-8°C until
centrifugation can be performed.
3) The blood must be processed by
centrifugation within 15 hours after
the blood was drawn.
4) Centrifuge the blood at 1000 x g for
15 minutes at 2-8°C.
5) After centrifugation, carefully collect
the serum (the straw-colored fluid)
by
pipetting
from
the
top
downwards.
6) Transfer the serum into two sterile,
labeled 1.5 ml microcentrifuge tubes,
aliquoting 0.5 ml into each tube.
7) Freeze the serum at 20°C or colder
until use for GLT testing.
Plasma Collection
1. Collect blood from the tail artery or
other suitable vein into a pre-labeled
plasma collection tube or container
(either EDTA or citrate). Do not use
heparin as an anti-coagulant.
2. Store the blood at 2-8°C until
centrifugation can be performed.
3. The blood must be processed by
centrifugation within 15 hours after
the blood was drawn.
4. Centrifuge the blood at 1000 x g for
15 minutes at 2-8°C.
5. After centrifugation, carefully collect
the plasma (the straw-colored fluid)
by
pipetting
from
the
top
downwards.
6. Transfer the plasma into two sterile,
labeled 1.5 ml microcentrifuge tubes,
aliquoting 0.5 ml into each tube.
7. Freeze the plasma at 20°C or colder
until use for GLT testing.
Test Procedure In Brief
1) Thaw the serum or plasma sample.
2) High speed centrifuge sample to
concentrate reactive fraction.
3) Extract CNA (after this step CNA
may be stored at 80°C until further
analysis).
4) Perform PCR on extracted CNA with
melting curve analysis.
5) Compare the melting curves of
controls to test samples.
6) Calculate the results.
Interpretation of Test Results
Non-Reactive
· A sample is considered non-reactive
when reaction does not exceed the
detection limit (defined as average N
