SMILE
Media QC
Media QC SOP
Author:
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Review by
Heidi Hanes
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Effective (or Post) Date:
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SMILE Approved by:
Review date
PRO67-A-01
9 Feb 09
JHH
Peggy Coulter
8-Feb-12
SMILE Comments: This document is provided as an example only. It must be revised to accurately reflect your lab’s
specific processes and/or specific protocol requirements. Users are directed to countercheck facts when considering
their use in other applications. If you have any questions contact SMILE.
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Pro6.7-A-01
Jan-06
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SMILE
Media QC
I acknowledge that I have read, understand and agree to follow this SOP.
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533561381
Date
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SMILE
Media QC
PERSONAL PROTECTIVE EQUIPMENT
Performance of this procedure will expose testing personnel to biohazardous material and possible
chemical hazards.
All specimens must be handled as potentially infectious material as outlined in the [your safety
manual].
The reagent(s) and/or chemical(s), which are used in this assay, may be hazardous to your health if
handled incorrectly. A brief listing of precautions for each chemical hazard is included in the reagent
section of this procedure.
More extensive information concerning the safe handling of the reagents and/or chemicals used in this
assay, as well as other important safety information, may be obtained by consulting the Material Safety
Data Sheet (MSDS) and [your hazardous materials manual], and[your safety manual]. Before
performing any part of this assay, the technologist must take any and all precautions and adhere to all
prescribed policies.
This procedure may expose you to:
[ ] Bloodborne pathogens
[ ] Airborne pathogens
[ ] Hazardous reagents
To perform this procedure, you must use:
[ ] Gloves
[ ] Face shield, or safety glasses with mask
[ ] Laboratory coat
[ ] Gown
[ ] Biological safety cabinet
[ ] Fume hood
[ ] Ventilator
[ ] Disinfectant following procedure:
[ ] Diluted bleach (1:10v/v solution made fresh daily)
[ ] Amphyl
[ ] Caltech Dispatch with Bleach
[ ] Miscellaneous, after Hospital Infection Control Department approval
Reference for spill/decontamination:
[ ] MSDS
[ ] Chemical hygiene plan
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SMILE
Media QC
PRINCIPLE:
The initial and final identification of bacteria depends on biochemical reactions that are
valid only after careful isolation techniques.
The ability to isolate pathogenic bacteria from clinical specimens can only be as effective
as the media used for culture.
MATERIALS:
A. All media and ingredients are purchased from reputable manufacturers in dehydrated
form or as prepared media.
B. A log is kept for each bottle of dehydrated media and each lot of prepared media,
recording the following:
1. name of the manufacturer
2. lot number
3. date of receipt
4. quantity received
5. expiration date
Preparation:
1. All media are prepared according to the manufacturers’ directions.
2. Ingredients, lot numbers, pH, and date of preparation are recorded on a media quality
control log sheet.
Performance Parameters:
All media must perform expected results as per NCCLS M22-T12.
Storage Requirements:
1. All media are stored according to manufacturers’ directions.
2. Media which exceeds the manufacturers’ expiration date are discarded.
3. Dispensed and cooled media are stored at 4˚ – 8˚ C.
PROCEDURE – STEPWISE:
1. The sterility of all media is checked by overnight incubation at 35˚ C.
2. Media is also checked for its overall appearance which is recorded in the quality control
QC log sheet.
3. The quality of each lot of media prepared in-house is tested by inoculation of ATCC
strains of organisms to produce positive and negative results before being used for
clinical purposes.
4. Use the following Tables included in the Appendices A, B, and C for specific
media/organism combinations and expected results.
5. Inoculation methods include:
a. Differential tubed media: inoculation of separate tubes with a known positive and
negative control organism
b. Selective media: inoculation of separate plates with an organism, which the media
is selective for, and with organisms for which the media is inhibitory.
c. Isolation media: inoculation of fastidious microorganisms whose growth should
be supported on the medium.
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SMILE
Media QC
REPORTING RESULTS:
All results are recorded on media quality control log sheets.
Procedures for Abnormal Results:
1. When media does not pass quality control, the media is discarded and recorded in the
failure log, noting corrective action taken.
2. The problem is investigated by the Q.C. technologist or Lead technologist of the
laboratory section.
3. Whenever possible, corrective measures will be instituted to avoid reoccurrences.
LIMITATIONS OF THE PROCEDURE:
Guidelines for establishing expiration dating:
1. The performance and expiration dating of culture media are affected by numerous factors
that may vary from lot to lot.
2. Factors adversely affecting the expiration dating and performance of prepared media
include:
a. overheating of basal ingredients
b. Using improper storage temperature.
c. Underfilling medium container
d. Exposing medium to light
e. Alternating the storage temperature of sealed media from room temperature to 2˚ to
8˚ C
f. Using small fill volume (less than 1.0ml) of tubed medium
g. Using loosely capped tube media
h. Storing plates in open or improperly sealed bags.
REFERENCES:
1. National Committee for Clinical Laboratory Standards, Quality Assurance for
commercially prepared microbiological culture media Approved Guideline, NCCLS
document M22-T12, 1996.
2. Clinical Microbiology Procedure Handbook, American Society of Microbiology,
Washington, DC, 2004.
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SMILE
Media QC
Appendix A
Prepared Media QC
Testing Plate Media/Tube Media
A label is attached to all lots of prepared media. A copy of this label must be dated and
saved for each type of media, and for each lot used. Control ATCC strains specified by NCCLS
has been tested on each batch of media meeting the “Quality Assurance for Commercially
Prepared Microbiology Culture Media”. BBL, REMEL and ANAEROBE Systems supply the
following rated media for use in the laboratory:
Prepared Agar
Abbreviation
1.
2.
3.
4.
5.
BAP
BBE
BBA
BCSA
Choc
TSA with 5% Sheep Blood
Bacteroides Bile Esculin
Brucella Blood Agar
Burkholderia Cepacia Selective Agar
Chocolate II
6. Chocolate II with Bacitracin
Choc.Bac.
7. Columbia CAN with 5% Sheep Blood
8. GC-Lect
CNA
GC
9. HBT Bilayer
HBT
10. Hemo ID Quad
Hemo
11. Haemophilus Test Medium
12. MacConkey
13. Mannitol Salt Agar
14. Mueller Hinton Agar
15. Mueller Hinton Agar +5% Sheep Blood
16. Regan Lowe Charcoal
HTM
Mac
Mann
MHA
MHA+SB
REG
17. Group A Selective Strep
SSA
18. XLD Agar
19. BCYE Differential
XLD
BCYE
20. BCYE PAC
21. BCYE PAV
PAC
PAV
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Tested for Growth
Neisseria meningitides
ATCC # 13090
Hemophilus influenza
ATCC # 49766
Neisseria gonorrhoeae
ATCC # 49226
Gardnerella vaginalis
ATCC # 14018
Hemophilus influenzae
ATCC # 49766
Hemophilus parahemlyticus
ATCC # 10014
Bordetella pertussis
ATCC # 8467
Streptococcus pyogenes
ATCC # 8135
Legionella pneumophilia
ATCC # 33155
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Media QC
Appendix B
In-House Prepared Plate Media Q.C.
Media
Positive- Growth
Negative- No Growth
Campy Agar
Campylobacter jejuni
ATCC # 29428
E. coli/S. aureus
25922/25923
CINN Agar
Yersinia enterocolitica
ATCC # 23715
E.coli
25922
Eugon Agar
E.coli/S.aureus/Ps.aeruginosa
ATCC # 25922/25923/10145
GV Agar
B.fragilis/P.melanogenicus
ATCC # 25285/25845
E.coli/S.aureus
25922/25923
BHI-4 Agar
Corynebacterium jeiikeium
ATCC # 43734
E.coli
25922
BHI-5 Agar
Enterobacter cloacae
ATCC # 13047
P.aeruginosa
10145
Potassium tellurite
Enterococcus faecalis
ATCC # 12984
E. solitarius
49428
Pseudomonas Agar
Pseudomonas aeruginosa
ATCC # 10145
E.coli
25922
Sheep Blood Amp 20 Aeromonas hydrophilia
ATCC # 7965
E. faecalis
12984
TCBS
M.morganii
8019
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Vibrio parahemolyticus
ATCC # 17802
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SMILE
Media QC
Media
Acetamide Slant
Appendix C
In-House Prepared Tube Media
Positive Control
Negative Control
Pseudomonas Aeruginosa
F. odoratum
ATCC #10145
ATCC #4651
Adonitol (Phenol Red)
C.diversus
ATCC#27156
C.freundii
ATCC#8090
Alkaline Peptone Water
A.hydrophila #7965
A.sobria #9071
P.shigelloides #14029
V.parahemolyticus #17802
uninoculated
Allantoin Slant- Discontinued C.testosteroni
ATCC #11996
P.aeruginosa
ATCC #10145
Arabinose (Phenol Red)
E.coli
ATCC #25922
S.marcescens
ATCC #8101
Arginine (Fay Berry)
P.aeruginosa
ATCC #10145
A.anitratum
ATCC #55489
Bile Esculin Agar
E.faecalis- #12984
S.bovis- #49147
S.viridans
ATCC #10556
CTA Lactose
N. lactamica
ATCC #23970
N.gonorrhoeae
ATCC #49226
CTA Maltose
N.meningitidis
ATCC #13090
P.multocida
ATCC #6529
CTA Mannitol
B.subtilis
ATCC #6051
N.gonorrhoeae
ATCC #49226
CTA Plain
N.gonorrhoeae- #49226
N.meningitidis- #13090
E.coli- #25922
S.aureus- #25923
CTA Raffinose
K.pneumoniae
ATCC #33495
P.mirabilis
ATCC #14153
CTA Salicin
A.hydrophilia
ATCC #7965
A.sorbia
ATCC #9071
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SMILE
Media QC
Media
CTA Sorbitol
Positive Control
E.faecalis
ATCC #12984
Negative Control
S.bovis
49147
CTA Sucrose
N.mucosa
ATCC #19695
M.catarrhalis
ATCC #8176
Dextrose (Phenol Red)
E.coli
ATCC #25922
P.aeruginosa
ATCC #10145
Dnase w/Methyl Green
S.marcescens- #8101
S.aureus- #25923
S.maltophilia- #12714
E.coli
ATCC #25922
Dulcitol (Phenol Red)
E.coli
ATCC #25922
M.morganii
ATCC #8019
Esculin Broth
E.faecalis- #12984
S.bovis- #49147
S.viridans
ATCC #10556
GN Broth
S.enteriditis- #31194
S.sonnei- #29930
E.coli
ATCC # 25922
HIB w/6.5%NaCl
E.faecalis
ATCC # 12984
S.bovis- #49147
S.viridans- #10556
Indole/Nitrate
E.coli
ATCC # 25922
K.pneumoiae- #33495
A.anitatum- #55489
Inositol (Phenol Red)
P.stuartii
ATCC #29914
C.freundii
ATCC # 8090
Lactose 10% agar slant
A.anitratum
ATCC # 55489
P.aeruginosa
ATCC #10145
Lactose (Phenol Red)
E.coli
ATCC # 25922
M.morganii
ATCC # 8019
Litmus Milk
C.perfringens- #3624
E.coli- #25922
Lysine (Fay Berry)
S.maltophilia- #12714
B.cepacia- #25416
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A.anitratum
ATCC # 55489
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SMILE
Media QC
Media
Malonate Broth
Positive Control
K.pneumoniae
ATCC # 33495
Negative Control
E.coli
ATCC # 25922
Maltose (Phenol Red)
K.pneumoniae
ATCC # 33495
M.morganii
ATCC #8019
Mannitol (Phenol Red)
K.pneumoniae
ATCC # 33495
M.morganii
ATCC # 8019
MIO Motility
Indole
Ornithine
E.aerogenes- #13048
E.coli- #25922
E.aerogenes- #13048
K.pneumonie
ATCC #33495
Arginine (Decarboxylase)
E.cloacae
ATCC # 13047
P.mirabilis
ATCC # 14153
Lysine (Decarboxylase)
K.pneumoniae
ATCC # 33495
P.mirabilis
ATCC # 14153
Ornithine (Decarboxylase)
E.aerogenes
ATCC #13048
K.pneumoniae
ATCC # 33495
Control Decarboxylase
E.aerogenes
ATCC #13048
P.aeruginosa
ATCC # 10145
MR/VP
M.morganii- #8019
H.alvei- #29826
H.alvei- #29926
M.morganii- #8019
Mueller Hinton Broth
E.coli- #25922
S.aureus- #25923
Nitrate Broth
E.coli- #25922
P.aeruginosa- #10145
P.multocida- #6529
A.anitratum- #55489
L.monocytogenes- #15313
Nutrient Broth
A.hydrophilia
ATCC # 7965
V.parahemolyticus
ATCC # 17802
Nutrient Broth w/6.5%
NaCl
V.parahemolyticus
ATCC # 17802
A.hydrophilia
ATCC # 7965
Dextrose (OF)
P.aeruginosa
ATCC # 10145
M.osloensis
ATCC # 10973
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SMILE
Media QC
Media
Maltose (OF)
Positive Control
S.maltophilia
ATCC # 12714
Negative Control
P.aeruginosa
ATCC # 10145
Xylose (OF)
P.aeruginosa
ATCC # 10145
A.lwoffi
ATCC #15309
Phenylalanine Slant
P.mirabilis
ATCC # 14153
E.coli
ATCC # 25922
Plain (Phenol Red)
E.coli- #25922
S.aureus- #25923
Pyruvate Broth
E.faecalis
ATCC # 12984
E.hirae
ATCC #8043
Raffinose (Phenol Red)
K.pneumoniae
ATCC # 33495
P.mirabilis
ATCC # 14153
Rhamnose (Phenol Red)
K.pneumoniae
ATCC # 33495
P.mirabilis
ATCC # 14153
Schaedler Broth
C.perfringens- #3624
H.influenzae- #49766
S.aureus- #25923
S.pyogenes- #6135
S.pneumoniae- 6303
SIM H2s
Indole
Motility
C.freundii-#8090
E.coli- #25922
E.aerogenes- #13048
K.pnuemoniae-#33495
E.aerogenes-#13048
K.pneumoniae-#33495
Simmons Citrate
K.pneumoniae
ATCC # 33495
E.coli
ATCC # 25922
Sodium Acetate
E.coli
ATCC # 25922
S.sonnei
ATCC # 29930
Sodium Hippurate
C.jejuni- #29428
S.agalactiae- #13813
G.vaginalis- #14018
S.pyogenes
ATCC #8135
Sorbitol (Phenol Red)
S.marcescens
ATCC # 8101
P.mirabilis
ATCC # 14153
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SMILE
Media QC
Media
Sucrose (Phenol Red)
Positive Control
K.pneumoniae
ATCC # 3395
Negative Control
M.morganii
ATCC # 8019
Todd Hewitt+Gentamicin
+ Nalidixic Acid
S.agalactiae
ATCC # 13813
E.coli- #25922
P.mirabilis- #14153
Trehalose (Phenol Red)
S.xylosus
ATCC #29971
S.epidermidis
ATCC #12228
Triple Sugar Iron (TSI)
E.coli- #25922
S.sonnei- #29930
P.mirabilis- #14153
P.aeruginosa- #10145
Trypticase Soy Agar
E.coli- #25922
P.aeruginosa- #10145
S.aureus- #25923
Trypticase Soy Broth
E.coli- #25922
S.aureus- #25923
Urea Agar Slants
P.mirabilis
ATCC #14153
E.coli
ATCC #25922
Xylose (Phenol Red)
P.mirabilis- #14153
S.xylosus- #29971
M.morganii- #8019
S.epidermidis- #12228
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