Quality Control in the Microbiology Laboratory-2

1. Temperature

2. Thermometer Calibration

3. Equipment Quality Control

4. Media Quality Control

5. Reagent Quality Control

6. Antimicrobial Susceptibility Quality Control

7. Personnel Competency

8. Use of Stock Cultures

9. Quality Control Manual

Methods of Sterilization-2

STERILIZATION

A. PHYSICAL METHODS:

1. Moist Heat – coagulation of bacterial proteins including bacterial enzymes

a. AUTOCLAVE – operates based on the principle of steam under pressure; effective indication:

STERILIZATION: 121oC for 15lbs/in2 for 15 minutes

DECONTAMINATION: 135oC for 30lbs/in2 for 30 minutes

biological indicator: Bacillus stearothermophilus

b, TYNDALLIZATION – fractional discontinuous sterilization; effective indication: 100oC for 30-60

minutes; instrument: Arnold’s Sterilizer
c. INSPISSATION – thickening through evaporation; effective indication: 75-80oC for 2 hours;
instrument: Inspissator............

2. Dry Heat – oxidation of bacterial components

a. DIRECT FLAME – direct application of flame in aseptic technique

b. DRY/HOT AIR OVEN – used in the sterilization of heat resistant materials; effective indication: 160-
180oC for 1.5 to 2 hours; biological indicator: Bacillus subtilis var. niger

c. INCINERATION – burns materials into ashes; used in the disposable of biological wastes; effective
indication: 870-980oC for 2 seconds

Ionizing Radiation- works by alkylation of nucleic acid of bacteria using high energy short wavelength
deep penetrating gamma rays; used for heat sensitive materials

biological indicator: Bacillus pumilis

4. Filtration – based on membrane gradient by differences in particle size

a. Water/Liquid solutions/Antibiotics

b. Air:

B. CHEMICAL METHODS:

1. Peracetic Acid
2. Formaldehyde Vapor/Vapor Phase H2O2

3. Glutaraldehyde

4. Ethylene Oxide (ETO) gas; biological indicator: Bacillus subtilis var. globijii

Antimicrobial Action-2

Antimicrobial Action:

A. Cell Wall inhibitors – sensitive for gram positive organisms

1. Penicillin – Penicillum notatum (gram pos)

2. Cephalosphorin – Cephalosporium (Acremonium)
3. Vancomycin – Streptomyces, treatment for MRSA (Penicillin R, Vancomycin S)
4. Aztreonam – Chromobacterium violaceum
5. Imipenem - carbapenems
6. Penicillinase Resistant Pen = Methicllin, Cloxacillin, Nafcillin, Oxacillin

B. Cell Membrance Inhibitors

1. Bacitracin
2. Colistin, Polymixin (Bacillus)
3. Amphotericin B, Nystatin (Streptomces), Imidazole, Clotrimazole (Anti-fungal)
 Structure destroyed by antibiotics: cell membrane (cell wall is not destroyed)

C. Ribosomes (CHON) Inhibitors – broad spectrum, for gram pos and gram neg

1. Aminoglycosides – gentamicin, kanamycin, tobramycin, netilmicin, amikacin, streptomycin

2. Tetracycline, chloramphenicol – bone marrow suppression
3. Erythromycin (macrolide)
 substitute for patients with allergy to penicillin
 eg.Clindamycin, Lincomycin

D. Nucleic Acid (DNA) Inhibitor

1. Mitomycin, Quinolones (levofloxacin, norfloxacin)
2. Metronidazole
 anaerobes, anti-protozoa, for amoebiasis
 G.vaginalis
 Ex. Flagyl
3. Sulfonamide-Trimethoprim (SXT) – inhibits folic acid (Ex. Rifampin)

Types Disinfectant-2

Disinfection/Antiseptic

Disinfection – non-sporocidal, some organisms are killed, kills pathogen

Disinfection for living things, Antiseptic for non-living things

1. 10% Sodium hypochlorite (Chorox)

 Spillage disinfectant
 10-30 minutes contact
2. Iodine/iodophor – sporocidal, best antiseptic for blood culture
3. 70% ethyl alcohol – non-sporocidal, cannot destroy viruses
4. 3-6% Hydrogen peroxide – cleansing of wound
5. 1% silver nitrate
 Crede’s prophylaxis
 Prevent gonococcal opthalmia neonatorium
6. Dyes – inhibits gram positive, anti-fungal
7. Formaldehyde - reject for bacterial culture because aldehyde is sporocidal
8. Glutaraldehyde (Cidex)
 cold sterilant
 for surgical equipment sterilization
9. Phenol (Carbolic Acid) - standard disinfectant
10. Lysol (Cresol)
11. Zephiran – benzalkonium chloride
12. Quats (Quaternary ammonium)
 Inactivated by organic materials

Note:
 Standard precautions - Blood and body fluids precautions must be observed for all patient’s blood and
body fluid specimen
 Universal precautions – all human blood and all other body fluids that contain visible blood
precautions must be observed