remel RapID ANA II System Kurzanleitung

RapID™ ANA II System
R8311002 .............................................................
1. INTENDED USE
The RapID™ ANA II System is a qualitative micromethod using
enzyme reactions to identify isolates grown on agar of clinically
significant anaerobic microorganisms. Used in a diagnostic
workflow to aid clinicians in treatment options for patients
suspected of having bacterial infections.
The device is not automated, is for professional use only and is
not a companion diagnostic.
The use of RapID ANA II System for identification and differentiation
of anaerobic bacteria of veterinary origin has not been fully
established. A complete listing of the organisms addressed by
RapID ANA II System is provided in the RapID ANA II Differential
Charts.
2. SUMMARY AND EXPLANATION
RapID ANA II System is comprised of (1) RapID ANA II Panels and
(2) RapID ANA II Reagent. RapID ANA II Panels are disposable
plastic trays with 10 reaction cavities, which contain dehydrated
reactants. The panel allows simultaneous inoculation of each
cavity with a predetermined amount of inoculum. A suspension
of the test organism in RapID Inoculation Fluid is used as the
inoculum which rehydrates and initiates test reactions. After
incubation of the panel, each test cavity is examined for reactivity
by noting the development of a color. In some cases, reagents
must be added to the test cavities to provide a color change. The
resulting pattern of positive and negative test scores is used as
the basis for identification of the test isolate by comparison with
the probability values in the Differential Charts (Table 4, 5 and 6),
or by use of the RapID ERIC™ software.
3. PRINCIPLE
The tests used in RapID ANA II System are based on microbial
degradation of specific substrates detected by various
indicator systems. The reactions employed are a combination
of conventional tests and single-substrate chromogenic tests,
described in Table 1.
4. REAGENTS
RapID ANA II Reagent (provided with kit)
Reactive ingredient per liter:
p-Dimethylaminocinnamaldehyde .....................................0.06 g
RapID Inoculation Fluid (R8325102, sold separately)(1 ml/tube)
KCl ......................................................................................6.0 g
CaCl .....................................................................................0.5 g
2
Demineralized Water ................................................... 1000.0 ml
RapID Spot Indole Reagent (R8309002, sold separately)
p-Dimethylaminocinnamaldehyde .....................................10.0 g
Hydrochloric Acid .......................................................... 100.0 ml
Demineralized Water ..................................................... 900.0 ml
5. WARNINGS & PRECAUTIONS
This product is for in vitro diagnostic use and should be used
by properly trained individuals. Precautions should be taken
against the dangers of microbiological hazards by properly
sterilizing specimens, containers, media, and test panels after
use. Directions should be read and followed carefully.
Non-disposable apparatus should be sterilised by any
appropriate procedure after use, although the preferred method
is to autoclave for 15 minutes at 121°C; disposables should be
autoclaved or incinerated. Spillage of potentially infectious
materials should be removed immediately with absorbent paper
tissue and the contaminated area swabbed with a standard
bacterial disinfectant or 70% alcohol. Do NOT use sodium
hypochlorite. Materials used to clean spills, including gloves,
should be disposed of as biohazardous waste.
Do not use reagents beyond the printed expiration dates.
Do not use if there is any evidence of contamination or other
signs of deterioration.
Any serious incident that has occurred in relation to the device
shall be reported to the manufacturer and the competent
authority of the Member State in which the user and/or the
patient is established. In the event of malfunction do not use
device.
Caution!
1. RapID ANA II Reagent is toxic and may cause harm to the
environment. Harmful by inhalation, contact with skin or
eyes, or if swallowed. May impair fertility or cause harm to
unborn child.
2. RapID Spot Indole Reagent may cause irritation to skin, eyes,
and respiratory system.
DANGER
H315 Causes skin irritation
H319 Causes serious eye irritation
H335 May cause respiratory irritation
H336 May cause drowsiness or dizziness
H360 May damage fertility. May damage the
unborn child
US ONLY
H373 May cause damage to organs through
prolonged or repeated exposure
P201 Obtain special instructions before use
P202 Do not handle until all safety precautions
have been read and understood
US & EU P281 Use personal protective equipment as
required
P264 Wash face, hands and any exposed skin
thoroughly after handling
P280 Wear eye/face protection
P260 Do not breathe dust/fume/gas/mist/
vapors/spray
P271 Use only outdoors or in a well-ventilated
area
P308+313 IF exposed or concerned: Get medical
attention/advice
P304+P340 IF INHALED: Remove victim to fresh air
and keep at rest in a position comfortable
for breathing
P302+P352 IF ON SKIN: Wash with plenty of soap
and water
P332+P313 If skin irritation occurs: Get medical
advice/attention
P362 Take off contaminated clothing and wash
before reuse
P305+P351 IF IN EYES: Rinse cautiously with water
+P338 for several minutes. Remove contact
lenses, if present and easy to do.
Continue rinsing
P337+P313 If eye irritation persists: Get medical
advice/attention
P405 Store locked up
P403+P233 Store in a well-ventilated place. Keep
container tightly closed
P501 Dispose of contents/container to an
approved waste disposal plant
EN 3. Refer to Safety Data Sheet, available on company website,
and product labeling for information on potentially hazardous
components, for detailed information on reagent chemicals.
Composition / information on ingredients
2-Methoxyethanol 109-86-4
Acetic acid 64-19-7
20 Hydrochloric acid 7647-01-0
WARNING! This product contains a chemical known in the State
of California to cause birth defects or other reproductive harm.
Emergency Telephone Number
INFOTRAC - 24 Hour Number: 1-800-535-5053
Outside of the United States, call 24 Hour Number: 001-352-
323-3500 (Call Collect)
6. STORAGE
Store RapID ANA II System and RapID Spot Indole Reagent in
their original containers at 2-8°C until used. Allow products to
equilibrate to room temperature before use. DO NOT interchange
reagents among different RapID systems. Remove only the
number of panels necessary for testing. Reseal the plastic pouch
and promptly return to 2-8°C. Panels must be used the same day
they are removed from storage. RapID Inoculation Fluid should
be stored in its original container at room temperature (20-25°C).
7. PRODUCT DETERIORATION
This product should not be used if (1) the expiration date has
passed, (2) the plastic tray is broken or the lid is compromised,
or (3) there are other signs of deterioration.
8. SPECIMEN COLLECTION, STORAGE, AND TRANSPORT
Specimens should be collected and handled following
recommended guidelines.
19,20
9. MATERIALS SUPPLIED
• 20 RapID ANA II Panels
• 20 Report Forms
• 1 RapID ANA II Reagent (one plastic dropper bottle containing
reagent sufficient for 20 panels)
• 2 chipboard incubation trays
• Instruction for Use (IFU)
• 1 color guide
10. CONTENTS SYMBOLS
ANA II Panels ANA II Panels
(15 ml/Btl)
RapID Report Forms RapID Report Forms
ANA II Reagent ANA II Reagent
Incubation Trays Incubation Trays
11. MATERIALS REQUIRED BUT NOT SUPPLIED
• Loop sterilization device
• Inoculating loop, swabs, collection containers
(15 ml/Btl)
• Incubators, alternative environmental systems
• Supplemental media,
• Quality control organisms
• Gram stain reagents
• Microscope slide
• Cotton swabs
• RapID Inoculation Fluid, 1 ml (R8325102)
• McFarland #3 turbidity standard (R20413) or equivalent
• Pipettes
• RapID Spot Indole Reagent (R8309002)
• ERIC (Electronic RapID Compendium, R8323600) (Optional).
12. PROCEDURE
Inoculum Preparation:
1. Test organisms must be grown anaerobically in pure culture
and examined by Gram stain prior to use in the system.
2. Test organisms may be removed from a variety of selective
and nonselective agar media. The following types of media
are recommended:
Nonselective Media: Anaerobic Blood Agar, Blood Agar
prepared with Brucella, Columbia or Tryptic Soy Base.
Differential or Selective Media: Egg Yolk Agar (EYA),
Kanamycin/Vancomycin (KV) Agar.
Notes:
• "Reducible agars" containing palladium chloride or other
reducing agents should NOT be used, as the reducing agents
may interfere with certain enzymatic activities.
• Kanamycin Bile Esculin (KBE) Agar and Bacteroides Bile
Esculin (BBE) Agar are NOT recommended since the ferric-
esculetin complex that may be formed can interfere with
test interpretation.
• Some media containing or supplemented with mono- or
disaccharides are NOT recommended since they may
suppress glycolytic activity and reduce test reactivity. Most
formulations of Schaedler agar contain sufficient dextrose to
interfere with glycosidase activity.
• Cultures used for inoculum preparation should be less than
72 hours old (preferably 18-24 hours old).
• 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 culture in RapID Inoculation Fluid
(1 ml) to achieve a visual turbidity equal to a #3 McFarland
turbidity standard or equivalent.
Notes:
• Suspensions significantly less turbid than a #3 McFarland
standard may result in aberrant reactions.
• Bacterial suspensions that are slightly more turbid than a
#3 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 #3 McFarland standard may compromise test
performance.
• Thoroughly mix suspensions and vortex if required.
• Use suspensions 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 anaerobically for at least 18-24 hours at 35-37°C.
Inoculation of RapID ANA II 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
test cavities at approximately 45° (see below).
Table 1. Principles and Components of the RapID ANA II Plus System
Cavity # Test Code Reactive Ingredient
Before Reagent Addition:
1 URE Urea
2 BLTS p-Nitrophenyl-β,D-disaccharide
3 αARA p-Nitrophenyl-α,L-arabinoside
4 ONPG o-Nitrophenyl-β,D-galactoside
5 αGLU p-Nitrophenyl-α,D-glucoside
6 βGLU p-Nitrophenyl-β,D-glucoside
7 αGAL p-Nitrophenyl-α,D-galactoside
8 αFUC p-Nitrophenyl-α,L-fucoside
p-Nitrophenyl-n-acetyl-β,D
9 NAG
glucosaminide
10 PO p-Nitrophenylphosphate
4
After Reagent Addition:
3 LGY Leucyl-glycine-β-naphthylamide 0.08%
4 GLY Glycine-β-naphthylamide
5 PRO Proline-β-naphthylamide
6 PAL Phenylalanine-β-naphthylamide 0.05%
7 ARG Arginine-β-naphthylamide
8 SER Serine-β-naphthylamide
9 PYR Pyrrolidonyl-β-naphthylamide
10 IND Tryptophane
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. 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.
Notes:
• Examine the test cavities. Test cavities 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.
Incubation of RapID ANA II Panels:
Incubate inoculated panels at 35-37°C in a non-CO2 incubator for
at least 4 hours but not more than 6 hours. For ease of handling,
panels may be incubated in the chipboard incubation trays
provided with the kit.
Note: If desired, after an incubation period of 4-6 hours and
prior to the addition of any reagents, RapID ANA II Panels may
be placed in the refrigerator (2-8°C) overnight for reading the
next morning.
Scoring of RapID ANA II Panels:
RapID ANA II panels contain 10 reaction cavities that provide 18
test scores. Test cavities 3 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 again after the addition
of reagent to provide the second test result. Bifunctional test
cavities 3 through 9, which require RapID ANA II Reagent, are
indicated with the first test above the bar and the second test
RapID ANA II Panel Test Location
Cavity # 1 2 3
Test Code URE BLTS αARA
LGY
Table 2. Interpretation of RapID ANA II System Tests*
Cavity # Test Code Reagent
Positive
Before Reagent Addition:
1 URE None Red or purple
2 BLTS
3 αARA
4 ONPG
5 αGLU
Medium or bright
6 βGLU None
yellow
7 αGAL
8 αFUC
9 NAG
10 PO
4
After Reagent Addition:
3 LGY
4 GLY
5 PRO
Purple, violet, red, or
6 PAL RapID ANA II Reagent
dark pink
7 ARG
8 SER
9 PYR
RapID Spot Indole
10 IND Blue or blue-green
Reagent
*NOTE: Panels should be read by looking down through the reaction cavities against a white background.
Quantity Principle
Hydrolysis of urea produces basic products
0.4%
which raise the pH and change the indicator.
0.1%
0.1%
0.1%
0.1%
Enzymatic hydrolysis of the colorless
0.08%
aryl-substituted glycoside or phosphoester
0.08%
releases yellow o- or p-nitrophenol.
0.08%
0.1%
0.1%
0.08%
Enzymatic hydrolysis of the arylamide
0.08%
substrate releases free β-naphthylamine
which is detected with RapID ANA II
0.05%
Reagent.
0.08%
0.08%
Utilization of the substrate results in the
0.01% formation of indole which is detected with
RapID Spot Indole Reagent.
below the bar. Bifunctional test 10, which uses RapID Spot Indole
Reagent, is designated with a box drawn around the reagent-
requiring test.
1. While firmly holding the RapID ANA II 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.
2. Without the addition of any reagents, read and score
cavities 1 (URE) through 10 (PO4) from left to right using
the color guide and the interpretation guide presented in
Table 2. Panels should be read by looking down through
the reaction wells against a white background. Record test
scores in the appropriate boxes on the report form using the
test code above the bar for bifunctional tests.
3. Add 2 drops of RapID Spot Indole Reagent to cavity 10 (IND).
Note: Only RapID Spot Indole Reagent should be used.
Kovacs' or Ehrlich's Indole reagent will not provide
satisfactory results.
4. Add 2 drops of RapID ANA II Reagent to cavities 3 (LGY)
through 9 (PYR).
5. Allow at least 30 seconds but no more than 2 minutes for
color development. Read and score cavities 3 through 10.
Record the scores in the appropriate boxes of the report
form using the test codes below the bar for bifunctional
tests.
6. Reference the microcode obtained on the report form in
ERIC for the identification.
13. RESULTS AND RANGE OF EXPECTED VALUES
The RapID ANA II Differential Charts (Tables 4, 5 and 6) illustrate
the expected results for the RapID ANA II 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
ANA II panels in conjunction with other laboratory information
(e.g., Gram stain, aerotolerance, growth on differential or
selective media, etc.) to produce a pattern that statistically
resembles known reactivity for taxa recorded in the RapID
System database. These patterns are compared through the
use of the RapID ANA II Differential Charts, or by derivation of a
microcode and the use of ERIC.
14. QUALITY CONTROL
All lot numbers of RapID ANA II System have been tested using
the following quality control organisms and have been found to
be acceptable. Testing of control organisms should be performed
in accordance with established laboratory quality control
procedures. If aberrant quality control results are noted, patient
results should not be reported. Table 3 lists expected results for
the selected battery of test organisms.
Notes:
• The quality control of RapID Reagents is accomplished by
obtaining the expected reactions for tests requiring the
addition of the reagents (cavities 3-10).
• Organisms which have been repeatedly transferred on agar
media for prolonged periods may provide aberrant results.
• Quality control strains should be stored frozen or lyophilized.
Prior to use, quality control strains should be transferred 2-3
times from storage on an agar medium that is recommended
for use with the RapID ANA II System.
4 5 6 7 8
ONPG αGLU βGLU αGAL αFUC
GLY PRO PAL ARG SER
Reaction
Comments
Negative
Shades of orange or reddish-orange
Yellow to orange
should be scored as negative.
Clear, tan, or very pale Only the development of a distinct
yellow yellow color is a positive test.
Allow at least 30 seconds but no more
than 2 minutes for color development.
Yellow, orange or pale
Only significant color development
pink
should be scored as positive. Pale
shades of color should be scored as
negative.
Any shade of blue or blue-green should
Any other color be scored as positive without regard to
intensity.
Bibliography #
1-3
4-8
6, 8-14
15, 16
9 10
NAG PO
4
PYR IND