remel RapID STR System Bedienungsanleitung Seite 2

ENGLISH
PROCEDURE
Inoculum Preparation:
1. Test organisms must be grown in pure culture, examined by Gram
stain, and tested for hemolysis prior to use in this system.
Note: Hemolysis is enhanced by incubation anaerobically or in 5-
7% CO .
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2. Test organisms may be removed from nonselective agar growth
media. The following types of media are recommended:
Tryptic Soy Agar (TSA) with or without 5% Sheep Blood; Nutrient
Agar; Chocolate Agar.
Notes:
•
Some media containing or supplemented with mono- or
disaccharides are not recommended since they may suppress
glycolytic activity and reduce test selectivity.
•
Plates used for inoculum preparation should preferably be 18-
24 hours old. Slow growing isolates may be tested using 48-
hour plates.
•
The use of media other than those recommended may
compromise test performance.
3. Using a cotton swab or inoculating loop, suspend sufficient growth
from the agar plate in RapID Inoculation Fluid (1 ml) to achieve a
visual turbidity equal to a #1 McFarland turbidity standard or
equivalent.
Notes:
•
Suspensions significantly less turbid than a #1 McFarland
standard may result in aberrant reactions.
•
Bacterial suspensions that are slightly more turbid than a #1
McFarland standard will not affect test performance and are
recommended for stock cultures and quality control strains.
However, bacterial suspensions significantly more turbid than a
#1 McFarland standard may compromise test performance.
•
Suspensions should be mixed thoroughly and vortexed if
required.
•
Suspensions should be used within 15 minutes of preparation.
4. An agar plate may be inoculated for purity and any additional testing
that may be required using a loopful of the test suspension from the
inoculation fluid tube. Incubate the plate for 18-24 hours at 35-37°C.
Inoculation of RapID STR Panels:
1. Peel back the lid of the panel over the inoculation port by pulling the
tab marked "Peel to Inoculate" up and to the left.
2. Using a pipette, gently transfer the entire contents of the Inoculation
Fluid tube into the upper right-hand corner of the panel. Reseal the
inoculation port of the panel by pressing the peel-back tab back in
place.
3. After adding the test suspension and while keeping the panel
on a level surface, tilt the panel back away from the reaction cavities
at approximately a 45-degree angle (see below).
4. While tilted back, gently rock the panel from side to side to evenly
distribute the inoculum along the rear baffles as illustrated below.
5. While maintaining a level, horizontal position (best achieved by using
the bench top against the reaction cavity bottoms), slowly tilt the
panel forward toward the reaction cavities until the inoculum flows
along the baffles into the reaction cavities (see below). This should
evacuate all of the inoculum from the rear portion of the panel.
Note: If the panel is tilted too quickly, air may be trapped at the test cavity
junction, restricting fluid movement.
6.
Notes:
•
•
•
Incubation of RapID STR Panels:
Incubate inoculated panels at 35-37°C in a non-CO
For ease of handling, panels may be incubated in the chipboard
incubation trays provided with the kit.
Scoring of RapID STR Panels:
RapID STR panels contain 10 reaction cavities that, in addition to
hemolysis, provide 15 test scores. Test cavities 7 through 10 are
bifunctional, containing two separate tests in the same cavity. Bifunctional
tests are first scored before the addition of reagent providing the first test
result, and then the same cavity is scored after the addition of reagent to
provide the second test result. The bifunctional test cavities that require
RapID STR Reagent are indicated with the first test above the bar and the
second test below the bar.
Cavity #
Test Code
1.
2.
3.
4.
Biochemical Wells
Inoculating Trough
(Back of Tray)
5.
6.
RESULTS AND RANGE OF EXPECTED VALUES
The RapID STR Differential Chart illustrates the expected results for
RapID STR System. Differential Chart results are expressed as a series of
positive percentages for each system test. This information statistically
supports the use of each test and provides the basis, through numerical
coding of digital test results, for a probabilistic approach to the
identification of the test isolate.
Identifications are made using individual test scores from RapID STR
panels in conjunction with other laboratory information (i.e. Gram stain,
hemolysis, colonial morphology, growth on differential or selective media) to
produce a pattern that statistically resembles known reactivity for taxa
recorded in the RapID STR System database. These patterns are
compared through the use of the RapID STR Differential Chart, or by
derivation of a microcode and the use of ERIC.
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Reaction Cavities
(Front of Tray)
Return the panel to a level position. If necessary, gently tap the
panel on the bench top to remove any air trapped in the cavities.
Examine the test cavities, which should appear bubble-free and
uniformly filled. Slight irregularities in test cavity fills are acceptable
and will not affect test performance. If the panel is grossly misfilled,
a new panel should be inoculated and the misfilled panel discarded.
Complete the inoculation of each panel receiving inoculation fluid
before inoculating additional panels.
Do not allow the inoculum to rest in the back portion of the panel for
prolonged periods without completing the procedure.
RapID STR Panel Test Location
1 2 3 4 5
ARG ESC MNL SBL RAF
While firmly holding the
RapID STR panel on the benchtop, peel off
the label lid over the reaction cavities by pulling the lower right hand
tab up and to the left.
Without the addition of the reagent, read and score cavities 1 (ARG)
through 10 (PO ) from left to right using the interpretation guide
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presented in Table 2. Record the test scores in the appropriate
boxes on the report form using the test code above the bar for
bifunctional tests.
Add 2 drops of RapID STR Reagent to cavities 7 (TYR) through 10
(PYR).
Allow at least 30 seconds but no more than 3 minutes for color
development. Read and score cavities 7 through 10. Record the
scores in the appropriate boxes of the report form using the test
codes below the bar for bifunctional tests.
Record the hemolysis reaction for the test isolate in the box provided on
the report form. The hemolysis reaction serves as the 15
should be scored as positive for beta-hemolytic isolates only. Alpha
and gamma hemolysis should be scored as negative.
Reference the microcode obtained on the report form in ERIC
(Electronic RapID Compendium) for the identification.
incubator for 4 hours.
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6 7 8 9 10
INU GAL GLU NAG PO
4
TYR HPR LYS PYR
th
test and