Transcript Slide 1

Presents
Sensi-Ring™
Antibiotic Sensitivity Detection System
Procedures and Techniques For Performing
In-house Antibiotic Sensitivities On Cultured
Kacey MultiChrome™ Bi-pates.
“Choosing the right drug for the right bug”
Why use Kacey Sensi-Ring™?

In house rapid identification (24 hours) of bacterial antibiotic
sensitivity using 8 of the most common veterinary antibiotic
choices per ring.

Cost Effective: Less than 1/3 the cost of a commercial laboratory
like Antech and Idexx.

Specific Sensi-Ring for both Gram Positive and Gram Negative

System specific Sensi-Ring:
- Urinary Tract Infection
- Ear Infection
- Skin and Wound Infection
Sensi-Ring™ Supplies For Each Test
For added convenience we also offer a Culture & Sensitivity Starter Kit
enough for four complete tests with all the supplies you need. Product #20241
Step 1: Obtaining live cultured bacteria
from a Kacey MultiChrome™ bi-plate.
•
Using the yellow loop, take 3 or 4 passes through
the bacterial growth on the MultiChrome™.
For bacterial colonies in both chambers see next slide.
Step 1 (con’t): Procedure For MultiChrome
bi-plates with bacteria on both the Gram
Positive and Negative side.
• In this case, you will prepare two Kacey Maxi Mueller
Hinton plates as indicated. One for the (+) and one for
the (-).
• To each MH plate you will add the appropriate Sensi-Ring
disk Example: place a Gram (+) in the MH plate with Gram (+) and
Gram (-) in the MH plate with Gram (-). Then incubate each.
Step 2: Mixing with Saline (WST) to dilute
•
Insert the yellow loop into the saline and twirl
to make a cloudy solution. This is an important
step to avoid overgrowth of a bacterial colony.
Step 3: Removing the diluted sample with
enclosed Rayon* Swab
•
Take a sample using the applicator tip of the swab.
* Do not use cotton swabs even if sterile. Cotton will release
a chemical that will alter the results.
Step 4: Inoculate diluted bacteria onto
Kacey Maxi* Mueller Hinton plate
•
Using the diluted bacteria solution on the soaked
swab, evenly distribute the sample onto a warmed
Kacey Maxi Muller Hinton plate (37 degrees 15-20 min).
• Streak the plate as shown
below. 11-5, 1-7, 9-3 o’clock
*Kacey Maxi Mueller Hinton plates are “true” 100mm plates. Most
commercially sold Muller Hinton plates are 88mm and will not work
with this system.
Step 5: Apply the Sensi-Ring™
• Pick the appropriate Sensi-Ring™ ie. Gram Positive,
Gram Negative, UTI or Skin/wound depending on
samples origin.
• Grasp the tab on the inside of the ring and lower
onto the agar upside down with lettering facing agar,
centering the ring with tweezers.
Step 6: Tamp down Sensi-Ring™
• Make sure that all of the disks touch the agar
plate to allow antibiotic diffusion.
Step 7: Incubate Upside Down
• Place Kacey Maxi Mueller Hinton plate now
inoculated upside down to prevent any contamination.
• Place into a preheated Kacey incubator at 37 degrees
for 24 hours.
24 hours
Step 8: Remove and read after 24 hours
• Carefully remove plate from incubator keeping it
upside down so that Sensi-Ring letters can be seen.
• Use gloves. Remember this is a live bacterial colony.
Step 9: Use the Kacey Zone Reader Card
• Place Kacey Maxi Mueller Hinton Plate upside down
with antibiotic letters facing up.
• Place Zone Reader over the plate and align the
notch of the Reader to the notch of the Sensi-Ring.
• Concentric circles should now encircle all of the disks.
• Read the Inhibition Zone in mm for each disk.
Notch
Step 10: Log Test Results
• Using the Patient Lab Report form supplied log
inhibition zones for each antibiotic disk.
Step 11: Treat with appropriate antibiotic
• Based on results, a course of action can now be
taken with the most efficacious antibiotic(s).
• The following is a list of commonly found Aerobic
Bacteria in small animal practice:
Gram Positive: Staphylococci, Streptococci, Enterococci
Gram Negative: E coli, Proteus, Klebsiella, Pseudomonas
Ear: Staphylococci, Pseudomonas, Proteus, E. coli, Enterococci, Streptococci
UTI: Staphylococci, Proteus, E. coli, Klebsiella, Enterococci, occasionally Pseudomonas
Skin & Wound: Staphylococci, Streptococci, Pseudomonas, Enterococci, Proteus
Step 12: Disposal of plate & clean up
• Once read, plate should be sprayed with a bactericidal
agent, covered with its lid taped shut, placed in a plastic
bag and disposed of in your bio-hazard bin.
• If you have a Kacey Sanitizer unit follow its instructions.
Place lid back on the plate, tape it shut, bag it and dispose
of it in your bio-hazard bin.
• Dispose of all used supplies in your bio-hazard bin.
• Spray your counters down and wash your hands in an
aseptic manor. Remember, these are live cultures.
Alternate Bacteriology Protocol Inoculating
Mueller Hinton Plate First Without Using the
MultiChrome Culture Plates
With Kacey Culture and Sensitivity system is it possible to
first directly inoculate a Maxi Mueller Hinton plate, incubate
for 24 hours and determine most efficacious antibiotic before
actually identifying the pathogenic bacteria.
A sample can then be drawn from the center area of the
Sensi-Ring™ on the Maxi Mueller Hinton plate and
transferred to a MultiChrome™ plate for incubation and
identification of the bacteria(s).
Alternate Bacteriology Protocol Inoculating
Mueller Hinton Plate First Without Using the
MultiChrome Culture Plates
•
Urine Samples: Spin down urine and do cytology on
the sample of the sediment.
- If bacteria is present, directly inoculate sediment
onto a Kacey Maxi Mueller Hinton plate with a
sterile Rayon swab enclosed smeared evenly
over the surface as described in step 4 .
• Ear Samples: Do a cytology on each ear swab.
- If bacteria is present, directly inoculate the Kacey
Maxi Mueller Hinton plate with sample smeared
evenly over the surface as described in step 4.
Sensi-Rings™ case samples
Case study: 2 year old canine which cultured on the MultiChrome™ both a
large colony of Gram Positive Staph in pink and a smaller Strep colony in blue.
The colonies when tested for sensitivity using the Sensi-Ring™ indicated
sensitivity to a variety of antibiotics not previously used.
Staph
Strep
UTI and Ear samples
Sensi-Rings™ Ability to identify
different strains of bacteria.
• Sensi-Rings™ used to identify antibiotic sensitivity
on different strains of cultured Staphylococci.
Sensi-Rings™ Quick Reference Card
Kacey Diagnostics Quick Reference For Culture & Sensitivity Study
1
INNOCULATION OF CULTURE BI-PLATE
1) Remove the plate from refrigerator & pre-heat
in the incubator @37 degrees for 15-20 minutes
prior to inoculating the Kacey MultiChrome biplate.
2) Using a sterile inoculating loop ( 20uL) for
liquids and a Kacey Sterile Swab for solids
,inoculate the sample onto both sides of the
Kacey Multichrome
Bi-plate, by utilizing a zigzag streaking motion.
3) Replace clear lid and place Kacey MultiChrome
Bi-plate in the incubator upside down (inverted
position) and incubate at 37°C +/- 2 degree C for
no less then 24 hours . The plate should be
examined after 24 hours, but no later than 48
hours after incubation.
GN
2
TRANSFER TO MULLER HINTON PLATE USING
KACEY “WST” TUBES
1) Taking a Kacey Sterile Swab carefully dab on to
three (3) different places containing the bacteria on
the MultiChrome Bi-plate.
2) Immediately place sterile Kacey Swab containing
the bacteria into a Kacey Working Solution tube
(WST containing 1.0 ml of 0.085%solution).
3) Mix the Kacey Swab with a gentle twirling motion
while in the WST tube for approximately 3-5
seconds. Use the same Kacey Swab for
inoculation of the Muller Hinton plate in step #3.
INOCULATION OF MULLER HINTON (MH) PLATE
1) Using the Kacey Swab containing the now diluted
WST dilution streak the 1st time the entire periphery
of the Mueller Hinton plate making a 360° circle
2) Streak a 2nd time again the Muller Hinton Plate
with a fresh sample of the WST dilution usingbroad
strokes starting at theto 3 o’clockposition.
2) Streak a 3rd time again the Muller Hinton plate
with a fresh sample of the WST dilution using wide
broad strokes starting at the 11 to 5 o’clock position
3) Remove “Sensi-Ring” from the foil pouch with
tweezers at the inner tab & place the “Sensi-Ring”
on to MH plate tapping down in non-disk areas, label
the specimen to be incubated.
4) Place the MH Inoculated plate into an incubator
upside down, set timer & incubate for 24 hours.
Remove & read inhibition zones with Kacey Digital
Reader.
GN
1
24 Hrs
2
GP
GP
3
WST
Dilution
3
New Kacey Sterile Swab
Quick reference only. Refer to detailed instruction manual for additional information.
Rotate Swab 3-5 sec
24 Hrs
Sensi-Rings™ Quick Reference Card (con’t)
Kacey Diagnostics
MICROBIAL ZONE INHIBTION
INTERPRETIVE CHART*
w/ Zone Diameter Interpretive Standards for
common veterinary Antibiotics
*These are the most widely accepted standard
procedures used by the US Food and Drug
administration (FDA) and the World Health
Organization (WHO). These standard procedure
values were adopted as a consensus standard by the
Clinical and Laboratory Standard Institute (CLSI)
formerly known as NCCL and are periodically
updated. The latest CLSI documentation should
always be consulted for current recommendations.
The list of abbreviations for the antibiotics has been
selected as standardization by KACEY® for
worldwide sales, distribution, nomenclature and
identification of the Kacey Sensi-Rings.
It is highly recommended that any bacterial agent
whose etiology and treatment is beyond the scope
of this system be sent to a qualified veterinary
reference lab for more detailed testing. It is also
recommended that any sample sent the lab be
obtained from the middle of the Sensi Ring on the
Kacey Maxi Muller Hinton Plate".
Antimicrobial Agent Name
Amikacin …………………………………………….
Amoxicillin …………………………………………..
Amoxi Clavulante …………………………………..
Augmentin-(Clavamox)…………………………….
Azithromycin-(Zithromax)………………………….
Cephalexin-(Keflex)………………………………..
Cefapodoxime ……………………………………..
Ceftazdime ………………………………………….
Ciprofloxacin –(Cipro)……………………………..
Clindamycin –(Clincaps)…………………………..
Cotrimoxazole (trimethoprim/sulfa)………………
Choloramphenicol-(Chloromycetin)………………
Cefalothin-(Keflin)………………………………….
Doxycycline-(Vibramycin)…………………………
Enrofloxacin-(Baytril)……………………………….
Gentamycin-(Gentocin)…………………………….
Neomycin …………………………………………...
Oxacillin ……………………………………………..
Penicillin……………………………………………..
Polymycin …………………………………………..
Ticarcillin-(Ticar)…………………………………….
Tobramycin-(Tobrex)……………………………….
Trimethoprim/Sulfa-(Bactrim Septra Primor)…….
Sensi Code
AK
A
AMC
AUG
ATH
CFX
CPD
CAZ
CIP
CD
TS
C
KF
DXT
ENF
GM
NE
OX
PG
PB
TC
TN
T/S
Disc Potency
30ug
30ug
30ug
30ug
13ug
30ug
10ug
30ug
5ug
30ug
25ug
30ug
30ug
30ug
5ug
10ug
30ug
1ug
10ug
300ug
75ug
10ug
25ug
Resistant
Intermediate
Susceptible
=14
=13
=13
=13
=13
=14
=17
=14
=15
=13
=10
=17
=14
=12
=17
=6
=12
=10
=28
=8
=14
=12
=10
15-16
14-17
14-17
14-17
14-17
15-17
18-20
15-17
16-20
14-17
11-15
18-20
15-17
13-15
18-20
7-9
13-16
11-12
——
9-11
15-19
13-14
11-15
=17
=18
=18
= 18
= 18
= 18
=21
=18
= 18
= 19
= 16
= 21
= 18
= 16
= 21
= 10
=17
=13
= 29
=12
= 20
= 15
= 16
Sensi-Rings™ & MultiChrome™ are trademarks Kacey® © 2013 Rev 10.3.2013 All Rights Reserved
1854 A Hendersonville Rd
Asheville, NC 28803
Ph: 1(828)685-3569
Fax: 1(828)685-3571
www.kaceyinc.org
“Kacey Products Providing Better Animal Care”
Kacey Diagnostics © 2014 All Rights Reserved
Rev 12.30.12
“MultiChrome”, “Sensi-Rings” are trademarks of Kacey® Inc.