Contributors

Terry Taff, MA

Rebecca Buxton, MS

MLS Micro 3

MicrobiologyMedical Laboratory Scientist

I. BacteriologyA. Basic principles

1. Differentiate among microorganisms (Level 1)a) Bacteriab) Fungi c) Virusesd) Parasitese) Prions

2. Describe the classification of bacteria (Level 1)a) Taxonomyb) Nomenclaturec) Identification

3. Describe the phenotypic characterization of bacteria (Level 1)a) Cell growth and reproductionb) Metabolism and nutrition

4. Describe the staining characteristics of bacteria (Level 1)a) Gram-positive, Gram-negative and Gram-variableb) Acid-fastness

5. Differentiate microscopic morphologies of bacteria and structures (Level 1)a) Cocci in chains, clusters, tetrads, pairsb) Diplococci and coccobacillic) Bacilli/rodsd) Unique shapes and arrangements

(1) Lancet(2) Fusiform(3) Pleomorphic(4) Branching(5) Palisading

e) Endosporesf) Capsulesg) Flagella

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h) Spirochetesi) Intra- and extra-cellular

6. Describe the use of bio- and molecular technology in taxonomy and clinical microbiology (Level 1)a) Deoxyribonucleic acid (DNA) relatednessb) Nucleic acid probes/hybridizationc) Amplification procedures including but not limited to

polymerase chain reaction7. Demonstrate proper use, maintenance and basic trouble-

shooting of the microscope (Level 2)B. Role of the clinical microbiology laboratory

1. Pre-analytical Phasea) Communicate with health professionals to insure quality

specimens for submission (Level 2)b) Recognize and resolve potential errors (Level 3)

2. Analytical Phasea) Accurately perform appropriate and timely testing in a

cost effective manner (Level 3)3. Post-analytical Phase

a) Provide accurate results (Level 3)C. Laboratory examination of specimens for bacterial culture

1. Properly identify specimen source (Level 2)a) CSFb) Blood and bone marrowc) Other body fluids

(1) Pleural(2) Synovial(3) Peritoneal(4) Pericardial(5) Amniotic (6) Gastric

d) Genitale) Upper respiratory

(1) Eye(2) Ear(3) Throat(4) Nasopharynx(5) Sinus

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f) Lower respiratory(1) Sputum(2) Bronchial

g) Skin and bone biopsiesh) Urine

(1) Catheterized(2) Clean voided midstream(3) Suprapubic

i) Wound(1) Abscess aspiration/purulent material(2) Surgical(3) Soft tissue

j) Stoolk) Autopsy

2. Provide proper accession of specimens (Level 2)a) Log inb) Request form/laboratory information system ordersc) Labeling

3. Determine acceptability of the specimen (Level 3)a) Rejection criteriab) Collection method/site preparation/aseptic techniquec) Collection timed) Container/Sampling devicee) Transport system, i.e., temperature, atmosphere, mediaf) Time in transitg) Patient therapyh) Number of specimensi) Optimum volumej) Contamination/spillage

4. Select appropriate storage temperature/environment if delay in processing (Level 2)

5. Select appropriate media and tests (Level 2)a) Proper routine primary isolation media

(1) Enriched/enrichment(2) Selective(3) Differential(4) Nutrient/general purpose

b) Purpose of each medium

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(1) Nutrients/constituents/supplements(2) Antibiotics(3) pH(4) Antibiotic removal(5) Environment

c) Specialized isolation mediad) Special stainse) Direct detection methods

6. Prepare specimen for inoculation (Level 2)a) Centrifugationb) Homogenization

7. Inoculate media (Level 2)a) Order of media for inoculationb) Quantitativec) Semi-quantitatived) Standard inoculation and streaking techniques

(1) Swab(2) Loop

(a) Reusable metal(b) Plastic(c) Calibrated

(3) Stab(4) Pipette(5) Automated plater

8. Incubate media (Level 2)a) Atmosphere

(1) Aerobic (ambient air) or microaerophilic(2) Carbon dioxide (CO2)

(a) 3-5%(b) 5-10% (capnophilic)

(3) Anaerobicb) Temperature

(1) 4o C(2) 25o C(3) 30˚C(4) 35o C(5) 42o C

c) Humidity

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d) Length of incubation9. Prepare and stain direct microscopic smears of specimen

(Level 3)a) Smear

(1) One-cell layer thick (may use cytocentrifugation of fluids)

(2) Air dry(3) Heat or methanol fix

b) Stains(1) Light microscopy

(a) Wet mounti. Saline

ii. Iodineiii. Colloidal carbon (India ink)iv. KOHv. Methylene blue

(b) Differentiali. Gram

ii. Spore(c) Acid-fast

i. Ziehl-Neelsenii. Kinyon

iii. Modified Kinyon(2) Fluorescent microscopy

(a) Acridine orange(b) Auramine-rhodamine(c) Calcofluor white(d) Fluorescein conjugated (FITC)

(3) Darkfield microscopy(4) Special

10. Evaluate and interpret microscopically (Level 3) a) Bacteria

(1) Structures(2) Capsule(3) Spores

b) Yeasts and hyphal elementsc) Red and white cellsd) Squamous and ciliated columnar epithelial cells i.e., clue

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cellse) Artifacts and background material

11. Quantitate organisms and cells (Level 2)12. Evaluate quality of specimen and assess clinical significance

of organisms present (Level 3)13. Differentiate normal flora from potentially pathogenic

organisms based on body site and specimen type (Level 3)D. Bacterial culture examination

1. Differentiate colony morphologies (Level 2)a) Staphylococcus spp.b) Streptococcus spp. and Enterococcus spp.c) Gram-negative coccid) Enterobacteriaceaee) Pseudomonas spp.f) Other non-fermentative Gram-negative rodsg) Fastidious and other miscellaneous Gram-negative rodsh) Non spore-forming Gram-positive rodsi) Spore-forming Gram-positive rodsj) Branching and filamentous Gram-positive rods

k) Mycoplasma spp. and Ureaplasma spp.l) Rapid growing Mycobacterium species

2. Differentiate common growth characteristics on routine media (Level 2)a) 5% Sheep Blood agar

(1) Hemolysis(a) Alpha(b) Beta

i. Double betaii. Subtle or narrow zone

(c) Gamma b) MacConkey, MacConkey with sorbitol (SMAC), eosin

methylene blue (EMB), hektoen-enteric (HE), xylose lysine desoxycholate (XLD), Salmonella-Shigella (SS)

(1) Fermenter and non-fermenter(2) H2S production(3) Lysine decarboxylation

c) Chocolate agard) Thayer Martin (TM), modified Thayer Martin (MTM),

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New York City (NYC), Martin Lewis (ML), JEMBECe) Campy-blood agar with cefoperazone, vancomycin,

amphotericin B (CVA)f) Colistin-nalidixic acid (CNA) blood agar, phenylethyl

alcohol (PEA), mannitol salt agar (MSA)g) Anaerobic blood agar, laked kananmycin-vancomycin

agar (LKV), Bacteroides bile-esculin agar (BBE)h) Trypticase soy broth, brain heart infusion broth,

thioglycollate plus vitamin K and hemin broth, Mueller-Hinton broth

i) Todd-Hewitt broth, LIM broth, Trans-vag broth, carrot broth

j) Gram-negative broth (GN)k) Campy brothl) Chromogenic agars

3. Select for, detect and recognize significant fastidious or unusual bacteria using selective and enrichment media (Level 3)a) Buffered charcoal yeast extract (BCYE) agarb) Cystine-tellurite and Tinsdale agarsc) Loeffler’s mediumd) Thiosulfate citrate bile salts sucrose (TCBS) agare) Cycloserine-cefoxitin fructose agar (CCFA)f) Yersinia selective agarsg) Bordet-Gengou and Regan-Lowe agarsh) Fletcher’s mediumi) Glucose-cysteine blood agarj) Burkholderia cepacia selective agar and oxidation

fermentation polymyxin bacitracin lactose agar (OFBL)k) Alkaline peptone water (APW)l) SP4 broth and agar

m) Shepard’s 10B brothn) A8 agar

4. Detect and recognize intracellular organisms using specialized isolation methods, e.g., shell vial and microwell plate assays for Chlamydia spp. and Chlamydiophila spp. (Level 2)

5. Evaluate growth on primary isolation media (Level 3)

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a) Correlation of direct gram stain results and culture results

b) Correlation of clinical diagnosis and specimen sourcec) Colony characteristics

(1) Size(2) Shape

(a) Elevation(b) Form(c) Margin--umbilicated, swarming, etc.

(3) Surface appearance(a) Mucoid(b) Transparent(c) Opaque, etc.

(4) Pigmentation(5) Odor(6) Changes in media

(a) Hemolysis(b) Pitting(c) Fermentation, etc.

d) Correlation of growth on various mediae) Differentiation of normal flora and potentially significant

isolatesf) Quantitation of growth

E. Identification of bacteria1. Apply principles of identification (Level 2)

a) Principlesb) Limitations and sources of errorsc) Troubleshootingd) Sensitivity and specificitye) Correlation of colony morphology and gram stain

reaction and microscopic morphologyf) Environmental requirements atmosphere, growth

temperature, etc.g) Metabolic rapid and traditional testing

(1) Catalase/superoxol(2) Oxidase/DMSO modified(3) Coagulase(4) Pyrrolidonyl arylamidase (PYR)

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(5) Leucine aminopeptidase (LAP)(6) CAMP factor(7) Hippurate hydrolysis(8) Esculin hydrolysis

(a) Rapid(b) Bile esculin slant

(9) Salt tolerance(10) Bile tolerance and solubility(11) TSI and KIA slants(12) H2S production(13) DNase/thermonuclease(14) Butyrate esterase(15) Growth factor requirement (X, V and XV)(16) Hemolysis on horse blood agar(17) Porphyrin (delta–aminolevulinic acid) (ALA)(18) Carbohydrate utilization

(a) Fermentation(b) Oxidation

(19) Beta-galactosidase (ONPG)(20) Indole

(a) Tube(b) Spot

(21) Methyl red and Voges Proskauer (MR/VP)(22) Citrate utilization(23) Phenylalanine deaminase (PAD)(24) Amino acid (ornithine and lysine) decarboxylase

and arginine dihydrolase using lysine iron agar (LIA)

(25) Urease(26) -glucuronidase (MUG)β(27) Nitrate reduction(28) Bile stimulation(29) Egg yolk agar

(a) Lipase(b) Lecithinase

(30) Gelatin hydrolysis(31) Indoxyl acetate(32) Starch hydrolysis

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h) Resistance or susceptibility to antibiotics or chemicals(1) Novobiocin(2) Bacitracin(3) Trimethoprim-sulfamethoxazole (SXT)(4) Optochin (ethylhydrocupriene hydrochloride)(5) Penicillin(6) Naladixic acid(7) Cephalothin(8) Special potency disks

(a) Colisitin(b) Kanamycin(c) Vancomycin

(9) Sodium polyanetholesulfonate (SPS)(10) Nitrofurantoin(11) Polymyxin B(12) Furazolidone(13) O/129

i) Other testing(1) Satellitism, i.e., Staphylococcus aureus streak(2) Motility(3) Elek test(4) Aerotolerance (5) Colony fluorescence(6) Halotolerance(7) String test

(a) KOH(b) Sodium deoxycholate

(8) Resistance to lysozyme(9) Hydrolysis of casein, xanthine, tyrosine

(10) Presence of aerial mycelium for Nocardia and Streptomyces

(11) Growth in presence of thionine and basic fuchsin dyes

j) Immunologic testing(1) Latex agglutination(2) Coagglutination(3) Urine antigen detection(4) Toxin detection

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(5) Immunofluorescent assays(a) Direct (DFA)(b) Indirect (IFA)

(6) Enzyme-linked immunoabsorbant assay (ELISA) (EIA)

(a) Solid phase(b) Membrane bound

(7) Serotypek) Commercial identification systems

(1) Non-automated(a) Miniaturized(b) Rapid

i. Substrate basedii. Spot tests

(2) Automated(3) Nucleic acid detection

(a) Nonamplifiedi. Hybridization probes

ii. Peptide nucleic acid probes with fluorescence in situ hybridization (PNA-FISH)

(b) Amplified including but not limited to real time polymerase chain reaction (PCR)

(4) Gas-liquid chromotography (GLC)(5) Matrix-assisted laser desorption ionization-time-

of-flight mass spectrometry (MALDI-TOF MS)2. Utilize algorithms and databases to establish identification

(Level 3)3. Isolate organisms (Level 2)4. Identify organisms and assess clinical significance (Level 3)

a) Staphylococcus spp.(1) Staphylococcus aureus

(a) Methicillin-resistant S. aureus (MRSA)(b) Vancomycin-intermediate S. aureus (VISA) (c) Vancomycin-resistant S. aureus (VRSA)

(2) Staphylococcus epidermidis(3) Staphylococcus saprophyticus(4) Staphylococcus lugdunensis

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(5) Other Staphylococcus spp.b) Micrococcus spp.c) Streptococcus spp. and Enterococcus spp.

(1) Streptococcus pyogenes (Group A)(2) Streptococcus agalactiae (Group B)(3) Other beta-hemolytic streptococci(4) Streptococcus pneumoniae(5) Viridans group

(a) S. anginosis(b) S. constellatus(c) S. intermedius

(6) Enterococcus(a) Vancomycin-resistant Enterococcus spp.

(VRE)(b) Enterococcus faecalis(c) Enterococcus faecium(d) Enterococcus gallinarum(e) Enterococcus casseliflavus

(7) Group D streptococcus(a) Streptococcus gallolyticus (previously S.

bovis)(8) “Nutritionally-variant streptococci (NVS)”

(a) Abiotrophia spp.(b) Granulicatella spp.

d) Other aerobic Gram-positive cocci(1) Leuconostoc spp.(2) Gemella spp.(3) Rothia mucilaginosa (4) Pediococcus spp.(5) Aerococcus spp.

e) Aerobic Gram-negative cocci(1) Neisseria gonorrhoeae(2) Neisseria meningitidis(3) Neisseria cinerea(4) Other Neisseria spp.(5) Moraxella catarrhalis

f) Enterobacteriaceae(1) E. coli

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(a) Enterohemorrhagic E. coli due to Shiga toxin

(b) Other diarrheagenic E. coli(2) Shigella spp.(3) Edwardsiella tarda(4) Klebsiella spp.

(a) K. pneumoniae(b) K. oxytoca

(5) Enterobacter spp. (a) E. aerogenes(b) E. cloacae

(6) Pantoea agglomerans(7) Serratia marcescens(8) Citrobacter spp.(9) Salmonella spp. and Salmonella enterica biovar

typhi(10) Proteus spp.

(a) P. mirabilis(b) P. vulgaris(c) P. penneri

(11) Providencia spp.(12) Morganella morganii(13) Yersinia spp.

(a) Y. pestis(b) Y. enterocolitica

(14) Plesiomonas shigelloides(15) Other Enterobacteriaceae

g) Other facultative Gram-negative rods(1) Vibrio spp.

(a) V. cholera(b) V. parahemolyticus(c) V. vulnificus(d) Other Vibrio spp.

(2) Aeromonas spp.(a) Aeromonas hydrophila(b) Other Aeromonas spp.

(3) Campylobacter spp.(a) C. jejuni

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(b) C. coli(c) Other Campylobacter spp.

(4) Helicobacter pylori(5) Chromobacterium violaceum(6) Others

h) Glucose nonfermenting Gram-negative rods(1) Pseudomonas aeruginosa(2) Other Pseudomonas spp.(3) Stenotrophomonas maltophilia(4) Burkholderia cepacia complex(5) Other Burkholderia spp.(6) Acinetobacter baumannii(7) Other Acinetobacter spp.(8) Alcaligenes faecalis(9) Achromobacter spp.

(10) Elizabethkingia meningoseptica (previously known as Chryseobacterium meningosepticum)

(11) Moraxella spp.(12) Shewenella spp.

i) HACEK and other fastidious Gram negative rods(1) Haemophilus parainfluenzae(2) Aggregatibacter aphrophilus (previously known

as Haemophilus aphrophilus and H. paraphrophilus)

(3) Aggregatibacter actinomycetemcomitans (previously Actinobacillus actinomycetemcomitans)

(4) Cardiobacterium hominis(5) Eikinella corrodens(6) Kingella spp.

j) Other Gram-negative bacilli/coccobacilli(1) Bartonella spp.

(a) B. quintana(b) B. henselae

(2) Bordetella spp.(3) Brucella spp.(4) Francisella tularensis(5) Haemophilus influenzae

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(a) Serotypes b and non-b(b) Biovar aegyptius

(6) Other Haemophilus spp.(7) Legionella spp

(a) Legionella pneumophila(b) Other Legionella spp.

(8) Pasteurella multocida(9) Capnocytophaga canimorsus

(10) Streptobacillus moniliformisk) Aerobic Gram-positive rods

(1) Gardnerella vaginalis(2) Corynebacterium diphtheriae(3) Corynebacterium jeikium(4) Corynebacterium urealyticum(5) Other Corynebacterium spp.(6) Listeria monocytogenes(7) Erysipelothrix rhusiopathiae(8) Bacillus anthracis(9) Bacillius cereus

(10) Other Bacillus spp.(11) Aerobic actinomycetes

(a) Nocardia spp.(b) Streptomyces spp.(c) Rhodococcus spp.

(12) Arcanobacterium haemolyticum l) Anaerobic Gram-positive rods

(1) Clostridium perfringens(2) Clostridium difficile(3) Clostridium septicum(4) Other Clostridium spp.

(a) C. tetani (b) C. botulinum

(5) Actinomyces israelii(6) Other Actinomyces spp.(7) Propionibacterium acnes(8) Lactobacillus spp.(9) Bifidobacterium spp.

(10) Eubacterium/Eggerthella spp.

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(11) Other anaerobic gram-positive rodsm) Anaerobic Gram-positive cocci

(1) Fingoldia magna (previously known as Peptostreptococcus magnus)

(2) Peptoniphilus spp. (previously known as Peptostreptococcus spp.)

(3) Peptostreptococcus spp.n) Anaerobic Gram-negative rods

(1) Bacteroides fragilis group(2) Other Bacteroides spp.(3) Fusobacterium spp.

(a) F. nucleatum(b) F. necrophorum

(4) Porphyromonas spp.(5) Prevotella spp.

(a) Prevotella melaninogenica(b) Other Prevotella spp.

o) Anaerobic Gram-negative cocci, i.e., Veillonella spp.p) Mycobacterium spp.q) Spirochetes

(1) Treponema pallidum subspecies pallidum(2) Non-venereal Treponema spp.(3) Borrelia spp.

(a) B. burgdorferi(b) B. recurrentis

(4) Leptospira interrogansr) Miscellaneous bacteria

(1) Mycoplasma spp.(a) M. pneumoniae(b) M. hominis

(2) Ureaplasma urealyticum(3) Chlamydia trachomatis(4) Chlamydophila spp.

(a) C. psittaci(b) C. pneumoniae

(5) Rickettsia rickettsii(6) Orientia tsutsugamushi (7) Ehrlichia spp.

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(8) Anaplasma phagocytophilum. (9) Coxiella burnetii

(10) Spirillum minus5. Utilize public health and reference laboratories for special

tests (Level 2)a) Laboratory resource informationb) Specimen handling

(1) Packaging and shipping regulations(2) Safety precautions(3) Transport conditions

c) Requisition informationd) Records/documentation/protocolse) Cost

F. Antimicrobials1. Apply standard performance principles and quality control to

antimicrobial susceptibility tests to insure accurate results (Level 3)a) Principlesb) Limitations and sources of errorsc) Troubleshootingd) Sensitivity and specificitye) Quality controlf) Disk diffusion (Kirby-Bauer) and antimicrobial gradient

method (E-test)(1) Media

(a) Depth(b) Cation content(c) Thymidine/thymine content(d) pH(e) Supplements(f) Storage

(2) Inoculum(a) Selection of appropriate organism for

method(b) Standardized suspension(c) Time limit for inoculation of media(d) Pattern of inoculation(e) Time limit for application of disks

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(f) Disk placement(3) Incubation

(a) Time(b) Temperature(c) Atmosphere

(4) Disk potency and storage(5) Reading(6) Interpretation

(a) Qualitative(b) Quantitative

(7) Reporting(8) Special techniques(9) Error detection and resolution

g) Beta-lactamase detectionh) Broth and agar dilution and automated systems

(1) Media(a) Cation content(b) Thymidine/thymine content

(2) Antibiotic solution(3) Inoculum(4) Selection of appropriate organism for method(5) Incubation(6) Reading(7) Interpretation of minimum inhibitory

concentration (MIC)(8) Reporting(9) Supplements and special techniques

(10) Error detection and resolutioni) Minimum bactericidal concentration (MBC)j) Synergism

k) Serum bactericidal titerl) Molecular methods to detect resistance genes

2. Utilize Clinical and Laboratory Standard Institute (CLSI) guidelines (Level 3)

3. Select appropriate antimicrobial agents for testing based on organism and specimen source (Level 2)

4. Perform susceptibility testing and special resistance detection methods on appropriate organisms (Level 3)

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a) Oxacillin resistance for Staphylococcus spp.b) Inducible clindamycin resistance for Staphylococcus,

beta-hemolytic Streptococcus spp. and Streptococcus pneumoniae

c) Vancomycin resistance for Staphylococcus and Enterococcus spp.

d) High level aminoglycoside resistance for Enterococcus spp.

e) Penicillin resistance for Streptococcus pneumoniaef) Extended spectrum beta-lactamases (ESBL) for

Enterobacteriaceaeg) ampC enzymes for Gram-negative rodsh) Carbapenemase resistant Enterobacteriaceae (CRE)

5. Interpret results (Level 3)a) Qualitativeb) Quantitative

6. Review susceptibility data and resolve unusual antimicrobial profiles (Level 3)

7. Utilize “predictor” antimicrobial agents to detect resistance (Level 2)

8. Recognize multidrug-resistant organisms (MDRO) (Level 2)9. Report data utilizing cascade and selective reporting (Level

2)10. Classify antimicrobial agents according to mode of action and

spectrum of activity (Level 2)11. Describe mechanisms of bacterial resistance (Level 2)12. Describe side and toxicity effects (Level 2)13. Recognize antimicrobials within each major class and by

generic and brand name. (Level 1)14. Apply standard performance principles to antimicrobial

assays of body fluids (Level 2)a) Specimenb) Collectionc) Processingd) Storagee) Information neededf) Limitations and sources of errorsg) Troubleshooting

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h) Sensitivity and specificityi) Quality controlj) Methods

(1) Bioassay(2) Direct detection

15. Interact with other professionals to recommend appropriate drugs for testing (Level 3)a) Antibiotic usage committeeb) Pharmacyc) Infectious disease cliniciansd) Hospital epidemiologist/infection prevention committeee) Antibiogram data

G. Results1. Prioritize reporting of direct smears (Level 3)2. Prepare culture reports and assure quality of results (Level 3)

a) Correlation of reported results with:(1) Direct Gram stain(2) Body site/specimen type(3) Patient history/population(4) Identification testing results(5) Susceptibility testing results(6) Clinical significance of organisms(7) Other significant information

b) Selective reporting of antimicrobials3. Report normal flora appropriately (Level 2)4. Designate preliminary or finalized status (Level 1)5. Recognize and resolve issues encountered (Level 3)6. Report cultures concisely, clearly and in a timely manner

(Level 2)7. Document work performed (Level 2)8. Insure appropriate charges are billed (Level 2)

II. MycologyA. Basic principles of mycology

1. Describe characteristics of fungi (Level 1)a) Classification, taxonomyb) Eukaryotic cellsc) Reproductiond) Growth requirements

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2. Recognize characteristics morphologic structures (Level 2)a) Yeast cellsb) Hyphae/pseudohyphae/mycelia

(1) Hyaline hyphae(2) Dematiaceous hyphae(3) Septate or true hyphae(4) Aseptate or non-septate hyphae

c) Conidiophoresd) Vesiclese) Phialidesf) Metulaeg) Micro/macroconidiah) Rhizoidsi) Sporangiphorej) Sporangium

k) Sporangiosporesl) Columella

m) Chlamydoconidian) Blastoconidiao) Arthroconidiap) Germ tubesq) Annelidsr) Cleistotheciums) Peritheciumt) Ascosporesu) Basidiosporesv) Zygospores

B. Laboratory examination of fungal specimens 1. Describe proper collection methods (Level 1)2. Discuss appropriate transportation and storage of specimen

(Level 1)3. Determine acceptability of specimen (Level 3)4. Select appropriate media for culture of fungal specimens

(Level 2)a) Primary isolation media

(1) Without antibacterial or antifungal agents(a) Sabouraud’s dextrose agar (SDA)(b) Brain heart infusion agar (BHI) plus sheep

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blood(c) Sabouraud dextrose with brain heart

infusion (Sabhi) with sheep blood(d) Potato dextrose agar (PDA)

(2) With antibacterial agents (chloramphenicol, ciprofloxacin, gentamicin, penicillin or streptomycin)

(a) SDA, BHI, Sabhi,i. With sheep blood

ii. Without sheep blood (b) PDA(c) Inhibitory mould agar (IMA)

(3) With antibacterial agents and antifungal agents (cyclohexamide)

(a) Mycosel or mycobiotic agar(b) BHI and Sabhi

(4) Selective for dimorphic fungi, e.g., yeast extract phosphate agar (YEPA)

(5) Dermatophyte test medium (DTM)(6) Mycosel or mycobiotic agar (7) Selective and differential for yeast, e.g.,

CHROMagar Candida5. Discuss the purpose of each medium preparation (Level 1)

a) pHb) Antibacterial agentsc) Antifungal agents

6. Inoculate media (Level 2)a) Aspirates, tissue and boneb) Blood and bone marrowc) CSF and other body fluidsd) Hair, skin and nailse) Upper and lower respiratoryf) Urine

7. Discuss proper incubation (Level 1)a) Atmosphereb) Temperaturec) Humidityd) Length of incubation and examination schedule

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8. Perform (Level 2) and interpret (Level 3) direct microscopic smears of fungal specimena) KOHb) Calcofluor whitec) India inkd) Gram staine) Acid fast f) Giemsag) Periodic acid schiff (PAS)h) Gomori methenamine silver (GMS)i) Hematoxylin and eosin (H&E) j) Fluorescent monoclonal antibody

9. Differentiate yeasts and moulds from bacteria on routine mycology media (Level 2)

10. Perform and interpret procedures for microscopic observation of fungi (Level 2)a) Germ tube b) Cornmeal/rice (chlamydoconidia agar)c) Ascospore agar d) Scotch tape preparation with lactophenol cotton blue

(LPCB)e) Slide cultures

11. Correlate patient history and clinical symptoms with rate of growth on media, colonial morphology and microscopic structures in order to identify fungi and assess clinical significance (Level 3)a) Yeasts

(1) Candida spp.(a) C. abicans(b) C. glabrata (c) C. tropicalis(d) C. krusei(e) C. parapsilosis(f) C. lusitaniae(g) C. dubliniensis(h) Other Candida spp.

(2) Cryptococcus spp. (a) C. neoformans

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(b) Other Cryptococcus spp.(3) Trichosporon spp.(4) Geotrichum spp.(5) Malassezia spp., i.e., M. furfur(6) Rhodotorula spp.(7) Saccharomyces spp.

b) Dimorphic moulds(1) Blastomyces dermatitidis(2) Coccidioides spp., i.e., C. immitis(3) Histoplasma capsulatum(4) Sporothrix schenckii(5) Paracoccidioides brasiliensis(6) Penicillium marneffei

c) Brightly colored/hyaline moulds(1) Aspergillus spp.

(a) A. fumigatus(b) A. flavus(c) A. niger(d) Other Aspergillus spp.

(2) Penicillum spp.(3) Fusarium spp.(4) Scopulariopsis spp.(5) Paecilomyses spp.(6) Acremonium spp.(7) Chrysosporium spp.(8) Sepedonium spp.(9) Other

d) Dematiaceous(1) Alternaria spp.(2) Curvularia spp.(3) Cladosporium(4) Cladophialophora spp.(5) Phialophora spp.(6) Fonsecaea spp.(7) Exophiala spp.(8) Hortaea (previously known as

Phaeoannellomyces werneckii)(9) Bipolaris spp.

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(10) Phoma spp.(11) Epicoccum spp.(12) Pseudoallescheria boydii(13) Scedosporium spp.(14) Wangiella spp.(15) Other

e) Dermatophytes(1) Microsporum spp.

(a) M. canis(b) M. gypseum(c) M. audouinii

(2) Trichophyton spp.(a) T. rubrum(b) T. mentagrophytes(c) T. tonsurans

(3) Epidermophyton floccosumf) Zygomycetes

(1) Rhizopus spp.(2) Mucor spp.(3) Rhizomucor spp.(4) Absidia spp.(5) Cunninghamella spp.(6) Syncephalastrum spp.

g) Other(1) Pneumocystis jiroveci(2) Others

12. Utilize test methodologies for fungi identification (Level 2)a) Principlesb) Limitation and sources of errorsc) Troubleshootingd) Sensitivity and specificitye) Quality controlf) Rapid and traditional testing

(1) Assimilation/fermentation(2) Temperature tolerance(3) Physiological tests for Actinomycetes(4) Nutritional studies for dermatophytes(5) Mould/yeast conversion

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(6) Niger seed (bird seed) agar/Caffeic acid disk(7) Converse medium(8) Biochemicals/enzymes

(a) L-proline aminopeptidase (PRO)(b) Beta-galactosaminidase (MUGAL)(c) Urea

(9) Wood’s lamp fluorescence(10) In-vitro hair perforation(11) Rice culture(12) Serological tests(13) Antigen detection methods

(a) Cryptococcal antigen(b) Exoantigen (immunodiffusion)

(14) Commercial methods(15) Molecular methods

13. Utilize databases and reference materials in identification of fungi (Level 2)

III. ParasitologyA. Taxonomy and terminology for categories of parasites

1. Describe the distinguishing characteristics of categories of parasites (Level 1)a) Nematodes (roundworms)

(1) Tissue and blood(2) Intestinal

b) Cestodes (tapeworms)c) Trematodes (flukes)d) Protozoan

(1) Amebae(2) Flagellates(3) Sporozoa

(a) Plasmodium spp.(b) Coccidia

(4) Microsporidia(5) Ciliates(6) Other

2. Define terms and describe life cycles (Level 1)a) Definitive hostb) Intermediate host

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c) Reservoir hostd) Vectore) Zoonosisf) Hermaphroditeg) Paroxysmh) Recrudescencei) Schizogonyj) Sporogony

k) Xenodiagnosis3. Recognize characteristic structures of adults, larvae, ova,

cysts, trophozoites, etc. (Level 2)a) Operculumb) Proglottidc) Scolexd) Genital primordiume) Chromatoidal bodyf) Glycogen vacuoleg) Karyosomeh) Nucleusi) Peripheral chromatinj) Uterine branches

k) Axostylel) Kinetoplast

m) Sheathn) Undulating membrane

B. Specimen collection and handling1. Determine specimen acceptability (Level 3)

a) Collection methodb) Collection and receipt timesc) Specimen storaged) Number of specimense) Presence of interfering or contaminating substancesf) Preservatives

(1) 10% formalin(2) Polyvinyl alcohol (PVA)(3) Schaudinn solution (mercury free)(4) Sodium acetate-acetic acid-formalin (SAF)(5) Less toxic single tube systems

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g) Rejection criteria C. Examination of specimens

1. Examine the fecal specimen macroscopically (Level 2)a) Colorb) Presence of blood and mucousc) Consistency

(1) Watery/loose(2) Semisolid(3) Formed

d) Presence of parasite structures (proglottids, adult, scolex, etc.)

2. Perform direct microscopic examination of specimen (Level 2)a) Proper use of the microscope, objectives, and light

source b) Use of ocular micrometer for measurement of size

calibrationc) Use of direct wet mounts with saline and iodine

preparationsd) Systematic examination of prepared slidee) Detection of parasites

3. Select and perform appropriate concentration methods and stains (Level 3)a) Principlesb) Limitations and sources of errorsc) Trouble-shootingd) Sensitivity and specificitye) Quality controlf) Concentration methods and stains

(1) Concentration methods(a) Formalin-ethyl acetate or alternate

solvents sedimentation(b) Zinc-sulfate flotation

(2) Permanent stained smears(a) Trichrome/modified trichrome(b) Iron-hematoxylin(c) Modified Kinyons (acid-fast)(d) Calcofluor white

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(e) Auramine O(3) Auto fluorescence with ultra-violet microscopy

g) Preparation of reagents and stains4. Describe the detection and differentiation of specific

parasites (Level 2)a) Immunoassays b) Nucleic acid assays

5. Detect and identify parasites (Level 2)a) Nematodes

(1) Intestinal (a) Ascaris lumbricoides(b) Strongyloides stercoralis(c) Hookworm

i. Necator spp.ii. Ancyclostoma spp.

(d) Trichuris trichiura(e) Enterobius vermicularis

(2) Blood and tissue(a) Trichinella spiralis(b) Wuchereria bancrofti(c) Brugia malayi(d) Loa loa(e) Mansonella spp.(f) Onchocerca volvulus(g) Dracunculus medinensis

(3) Other(a) Anisakis spp.

b) Cestodes(1) Taenia solium(2) Taenia saginata(3) Echinococcus granulosus(4) Diphyllobothrium latum(5) Hymenolopis nana(6) Hymenolopis diminuta(7) Dipylidium caninum

c) Trematodes(1) Paragonimus westermani(2) Fasciolopsis buski

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(3) Fasciola hepatica(4) Clonorchis sinensis(5) Heterophyes heterophyes(6) Metagonimus yokogawai(7) Schistosoma mansoni(8) Schistosoma haematobium(9) Schistosoma japonicum

d) Protozoa(1) Amoebae

(a) Entamoeba histolytica(b) Entamoeba dispar(c) Entamoeba coli(d) Other Entamoeba spp.(e) Iodamoeba bütschlii(f) Endolimax nana (g) Blastocystis hominis(h) Naegleria fowleri(i) Acanthamoeba spp.

(2) Flagellates(a) Dientamoeba fragilis(b) Chilomastix mesnili(c) Giardia lamblia/intestinalis(d) Trichomonas vaginalis(e) Trypanosoma spp.(f) Leishmania spp.

(3) Sporozoa(a) Plasmodium falciparum(b) Plasmodium malariae(c) Plasmodium ovale(d) Plasmodium vivax(e) Plasmodium knowlesi(f) Babesia spp.(g) Toxoplasma gondii(h) Sarcocystis spp.(i) Cystoisospora belli (previously Isospora

belli)(j) Cryptosporidium parvum

(k) Cyclospora spp.

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(4) Ciliates(a) Balantidium coli

(5) Other(a) Microsporidia

6. Correlate information in order to identify parasites and assess clinical significance (Level 3)a) Diagnostic stage, i.e., characteristic structure(s) presentb) Knowledge of life cyclec) Specimen of choice for detectiond) Detection methods available

7. Differentiate artifacts from parasites (Level 2)a) White and red blood cellsb) Epithelial cellsc) Pollen granulesd) Plant fibers and cellse) Yeast cellsf) Charcot-Leyden crystalsg) Fungal spores h) Diatomsi) Hair

8. Examine specimens other than stool (Level 2)a) Cellophane tape/vaspar paddle preparation for

Enterobius vermicularisb) Wet mount/culture for Trichomonas speciesc) Duodenal capsule or string technique (Entero-Test)d) Thick and thin blood filmse) Bone marrow and body fluidsf) Urineg) Lower respiratoryh) Biopsy

9. Detect and recognize common arthropods (Level 2)a) Hard and soft body ticksb) Licec) Mitesd) Fleas

IV. MycobacteriologyA. General characteristics

1. Describe the general characteristics of the genus

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Mycobacterium (Level 1)a) Acid-fastnessb) Growth requirementsc) Rate of growthd) Optimum temperature and atmosphere

B. Specimen management1. Identify the safety requirements for working with

mycobacteria (Level 1)a) Biological safety cabinet (BSC)/Biosafety level (BSL)b) Personal protective equipment

(1) Respirator(2) Gloves(3) Liquid impervious gowns(4) Centrifuges with safety carriers(5) Germicides

c) Equipment d) Negative pressure facilitye) Annual tuberculin skin test

(1) Chest x-ray if skin test is positive(2) Effects of BCG vaccine

2. Describe specimen collection and transportation procedures (Level 2)a) Pulmonary sites

(1) Sputum, expectorated and induced(2) Bronchial alveolar lavage (BAL), bronchoscopy,

etc.b) Extrapulmonary sites

(1) Non-contaminated(a) Blood and bone marrow(b) Body fluids(c) Tissue

(2) Contaminated(a) Urine(b) Skin lesions, wound, abscesses(c) Gastric lavage or aspirate(d) Stool for Mycobacterium avium complex

(3) Blood for Interferon gamma release assay (QuantiFERON®-TB test)

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c) Collection method/site preparationd) Containere) Collection timef) Number of specimensg) Qualityh) Optimum volumei) Rejection criteria

3. Describe and utilize specimen processing procedures (Level 2)a) Contaminated specimens

(1) Digestion and decontamination(a) Liquefication

i. n-acetyl-l-cysteine (NALC)ii. Dithiothreitol

iii. Other(b) Decontaminaton

i. 4% NaOHii. Oxalic acid

iii. Other(c) Neutralization

i. Phosphate bufferii. Other

(d) Centrifugationi. Speed

ii. Timeiii. Equipment required

(e) Limitations and potential sources of errors(f) Quality assurance, i.e., maintenance of a

contamination rate of < 3-5%b) Non-contaminated specimens

(1) Centrifugation(2) Direct inoculation

C. Smears and stains1. Prepare and stain smears (Level 2)

a) Specimen selection(1) Smear preparation, standardization, fixation

(a) Direct(b) Concentrated

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(c) Cytocentrifugation with bleach b) Stains

(1) Reagent preparation(2) Acid- fast stain procedures

(a) Principlei. Flourochrome

(1) Auramine-rhodamine(2) Auramine-O

ii. Fuchsin based(1) Ziehl-Neelsen (hot)(2) Kinyoun (cold)(3) Modified Kinyoun

(b) Quality control(c) Limitations and potential sources of errors(d) Troubleshooting(e) Sensitivity and specificity

c) Microscopic evaluation(1) Magnification(2) Scanning pattern(3) Organism appearance, e.g., serpentine cording(4) Specificity(5) Sensitivity

2. Interpret and report stained smears (Level 3)a) Sources of false positives

(1) Nocardia(2) Rhodococcus(3) Cryptosporidium , Cystoisospora and Cyclospora

oocysts(4) M. gordonae from a tap water source(5) Other

b) Appearance of artifacts, debris, backgroundc) Reporting schemed) Internal review process for quality assurance

D. Culture medium1. Describe the culture media most appropriate for primary

cultures by specimen type (Level 1)a) Egg-based

(1) Lowenstein-Jensen

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(2) Petragnani(3) American Trudeau Society medium(4) Others

b) Agar-based(1) Middlebrook 7H10 and 7H11(2) Others

c) Liquid based (1) Middlebrook 7H9, 7H12 and 7H13(2) Dubois Tween albumin broth

d) Media supplemented with hemin or hemoglobine) Commercial systemsf) Other

2. Discuss the incubation of the primary media (Level 1)a) Describe optimal temperature to isolate mycobacteria

(1) 350- 370 C(2) 250- 330 C

b) Describe the optimal atmospheres of incubationc) Describe the optimal length of incubation d) Describe the reading schedule for inoculated media

E. Identification1. Identify isolates (Level 2)

a) Acid-fastness of the organismb) Preferred temperature of growthc) Growth requirements, i.e., hemind) Historic significance of Runyon classificatione) Rate of growth

(1) Rapid grower (< 7 days)(2) Slow grower (> 7 days)

f) Colony morphology(1) Pigment

(a) Colori. White

ii. Buffiii. Yellowiv. Orangev. Other

(b) Photoreactivity/photochromogenicity studies

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i. Photochromogenii. Scotochromogen

iii. Nonphotochromogen(2) Texture

(a) Smooth(b) Rough(c) Cording(d) Wrinkled/nodular(e) Mucoid(f) Other

F. Biochemical testing1. Select, perform, and interpret biochemical testing in order to

differentiate organisms (Level 3)a) Principleb) Interpretationc) Preparation of reagentsd) Quality controle) Limitations and potential sources of errorsf) Troubleshootingg) Sensitivity and specificityh) Biochemical testing

(1) Arylsulfatase(2) Catalase

(a) Semiquantitative (b) 680 C (heat-stable)

(3) Urease(4) Growth on MacConkey agar without crystal violet(5) Iron uptake(6) Niacin accumulation(7) Nitrate reduction(8) Pyrazinamidase(9) Sodium chloride tolerance

(10) Tellurite reduction(11) Tween 80 hydrolysis(12) Other

i) Other methods for organism identification(1) Gas liquid chromatography(2) High pressure liquid chromatography (HPLC)

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(3) Molecular diagnostics(4) Amplification methods for direct detection of

Mycobacterium tuberculosis2. Differentiate and identify mycobacteria based on key criteria

(Level 3)a) Mycobacterium tuberculosis complex

(1) M. tuberculosis(2) M. bovis(3) M. africanum

b) Mycobacterium avium-intracellulare complex (MAC or MAI)

c) Mycobacterium ulceransd) Mycobacterium haemophilume) Mycobacterium xenopif) Mycobacterium kansasiig) Mycobacterium marinumh) Mycobacterium gordonaei) Mycobacterium scrofulaceumj) Mycobacterium fortuitum complex

k) Mycobacterium chelonael) Mycobacterium abscessus

m) Mycobacterium leprae3. Correlate the presence of organisms with the most common

types of clinical infections and clinical significance (Level 3)a) Routes of transmissionb) Signs and symptoms

G. Susceptibility testing1. Describe the methods used for antimycobacterial drug

susceptibility testing and assay determinations (Level 1)a) Agar proportionb) Radiometricc) Commercial systems

2. Describe the rationale for multiple drug susceptibility testing (Level 1)

H. Reporting1. Report results (Level 2)2. Describe the turnaround time and reporting of direct smear,

culture, and susceptibility results (Level 1)

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3. Discuss the reporting of the final culture and susceptibility results (Level 1)

V. VirologyA. Characteristics of viruses

1. Describe the basic structure/components of viral agents (Level 1)a) Virion

(1) Type of nucleic acid present (RNA or DNA)(2) Capsid (3) Envelope(4) Glycoprotein spikes

2. Differentiate viruses from bacteria (Level 1)a) Requirement for living cellsb) Sizec) Structured) Replicatione) Therapy

B. Classification of viruses1. Outline the criteria for classifying or grouping viruses (Level

1)a) Nucleic acid type (RNA or DNA)b) Size and morphologyc) Chemical and physical characteristicsd) Means of replicatione) Host

2. Correlate agents of viral infection with disease or pathologic manifestations (Level 2), route of transmission (Level 2), appropriate diagnostic tests (Level 2) and assess clinical significance (Level 3)a) Coronavirusb) Human papillomavirus 6, 11, 16 and 18c) Eastern and western equine encephalitis virusd) Arenavirus

(1) Lassa virus(2) Lymphocytic choriomeningitis virus

e) SARS related coronavirusf) Cytomegalovirusg) Human T-lymphocytic virus

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h) Ebolavirusi) Enterovirus

(1) Poliovirus(2) Coxsackievirus(3) Enterovirus(4) Rhinovirus(5) Echovirus

j) Parvovirus B19k) Flavivirus

(1) West Nile virus(2) St. Louis encephalitis virus(3) Dengue virus(4) Yellow fever virus

l) Hantavirusm) Hepatitis viruses

(1) Hepatovirus, Hepatitis A virus(2) Orthohepadnavirus, Hepatitis B virus(3) Hepacivirus, Hepatitis C virus(4) Deltavirus, Hepatitis D virus(5) Hepevirus, Hepatitis E virus

n) Herpes viruses(1) Kaposi’s sarcoma associated herpesvirus(2) Simplex virus, Herpesvirus 1 and 2(3) Herpesvirus 6 and 7

o) Influenzavirus A(1) H1N1(2) H5N1

p) Influenzavirus Bq) Influenzavirus Cr) Rabies viruss) Human immunodeficiency virust) Epstein-Barr virusu) Marburgvirusv) Adenovirus

w) Metapneumovirusx) Measles virusy) Norwalk virusz) Hepatitis B virus

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aa) Rift valley fever virusbb) Respiratory syncytial viruscc) Parainfluenza virus

dd) Rotavirusee) Rubella virusff) Mumps virus

gg) Varicella-zoster virusC. Collection and processing of specimens

1. Describe appropriate collection and transport of specimens (Level 2)a) Selection of body site/specimen typeb) Collection methods, devices and containers c) Transport mediad) Time/temperaturee) Safety precautions

2. Utilize proper specimen storage upon receipt in laboratory (Level 1)

3. Utilize proper specimen shipment (Level 1)4. Utilize specimen processing algorithms based on most likely

virus present (Level 1)a) Rejection criteriab) Specimen type/body sitec) Specimen preparationd) Age of patiente) Time of year/virus seasonalityf) Virus suspectedg) Immune status

D. Laboratory detection of viral agents1. Perform (Level 2), interpret (Level 3), and describe (Level 1)

procedures for detection and identification of viruses, viral particles or viral antigensa) Principleb) Interpretationc) Preparation of reagentsd) Quality controle) Limitations and potential sources of errorsf) Troubleshootingg) Sensitivity and specificity

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h) Direct detection methods(1) Electron microscopy (EM)(2) Cytohistopathology(3) Immunodiagnostic

(a) Direct and indirect immuofluorescent, i.e., antibody methods

(b) Enzyme immunoassay methods (EIA)(ELISA)

(c) Latex agglutination(d) Immunoperoxidase

(4) Radioimmunoassay(5) Molecular methods

i) Cell Culture(1) Conventional monolayer

(a) Growth and maintenance media(b) Cytopathic effect (CPE)(c) Types of cell lines

i. Primaryii. Diploid or low passage

iii. Heteroploid or continuous(d) Blind passage(e) Identification of viruses

i. Rate of growth, i.e., characteristic CPE on specific cell line(s)

ii. Hemadsorptioniii. Confirmation by fluorescent antibody

stain(2) Other cell culture systems

(a) Shell vial(b) Microwell (cluster) plate(c) Cryopreserved monolayer(d) Co-cultured cells(e) Transgenic cell line, i.e., enzyme-linked

virus inducible system (ELVIS)j) Serology

VI. Infection prevention and controlA. Disease transmission

1. Define terms (Level 1)

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a) Epidemiologyb) Community acquired infectionsc) Nosocomial infectionsd) Epidemice) Endemicf) Outbreakg) Clusterh) Surveillancei) Morbidityj) Mortality

2. Describe the origin and mode of spread (Level 1)a) Dropletb) Airbornec) Fomited) Vectore) Reservoirf) Endogenousg) Exogenous

3. Compare and contrast (Level 2)a) Colonizationb) Infectionc) Carrier state

B. Infection prevention methods1. Relate underlying patient condition/factors to acquisition of

infection (Level 1)a) Medical devices (catheter, respirator)b) Immunocompromised statec) Immunosuppressive therapyd) Antimicrobial therapye) Malignancyf) Ageg) Occupationh) Surgeryi) Prosthetic devices (pacemaker, artificial heart valve,

shunt, joint)2. Apply concepts of disease transmission to disease prevention

(Level 2)a) Education

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(1) Health care professionals(2) Patients(3) Environmental services (housekeeping)

b) Precautions(1) Standard(2) Transmission-based

(a) Direct and indirect contact(b) Droplet(c) Airborne

c) Immunizationsd) Treatment

(1) Antibiotics(2) Antiviral(3) Antifungal(4) Antiparasitic

C. Role of clinical microbiology laboratory1. Culture and detect microbial pathogens (Level 2)

a) Common bacteriab) Multiple drug resistant organism (MDRO)c) Mycobacteriad) Fungie) Unusual organismsf) Virusesg) Parasites

2. Report relevant cultures/organisms to infection prevention personnel (Level 2)

3. Assist epidemiologist/hospital infection prevention personnel with surveillance and control of infectious diseases (Level 3)a) Environmental samples

(1) Water(2) Food(3) Air(4) Vectors/fomites(5) Operating room(6) Respiratory therapy equipment(7) Physical therapy equipment(8) Dialysis machines

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b) Personnel specimensc) Patient specimensd) Rapid diagnostic testinge) Organism identificationf) Antimicrobial susceptibility testingg) Epidemiologic analysis of microorganisms

(1) Phenotypic techniques(a) Biotyping(b) Antimicrobial susceptibility patterns(c) Serotyping(d) Bacteriophage typing

(2) Genotypic techniques(a) Plasmid analysis(b) Restriction endonuclease analysis (REA) of

chromosomal DNA(c) Southern blot analysis(d) Pulsed field gel electrophoresis (PFGE) of

chromosomal DNA(e) Molecular sequencing(f) Other

4. Report communicable diseases/organisms to the appropriate public health agencies (Level 1)

5. Monitor for bioterrorism agents and emerging infections (Level 2)a) Centers for Disease Control (CDC) categories of

organismsb) CDC laboratory response networkc) Specimen packing and shippingd) Biosafetye) Protocols to rule in/rule out critical agents

6. Maintain adequate archival information (Level 2)a) Data

(1) Health Insurance Portability and Accountability Act of 1996 (HIPAA)

b) Organisms(1) Patient(2) Personnel(3) Surveillance/environmental

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7. Handle and dispose of bio hazardous materials (Level 2)a) QC of autoclavesb) Identification of biological, pathological, and surgical

infectious materialsc) Cleaning, sterilization, disinfectiond) Laboratory safety procedures manuale) Aseptic technique

VII. Laboratory PracticeA. Quality management

1. Create and maintain policies and procedures (Level 3)2. Perform procedures (Level 2) and evaluate results (Level 3)

of standard quality controla) Mediab) Stainsc) Reagents/kitsd) Equipmente) Physiological testsf) Antimicrobial testingg) Serological testsh) Stock organismsi) Inventoryj) Nucleic acid probes

k) Nucleic acid amplification methodsl) Automated systems

m) Immunological testn) Microscope calibrationo) Cell culture lines

3. Recognize and resolve errors (Level 3) 4. Create and implement a quality management plan (Level 3)5. Identify significant risks to quality and patient safety (Level

2)6. Create controls and monitors to reduce risk (Level 3)7. Monitor performance (Level 2)

a) Complaints, incidents and sentinel eventsb) Benchmarksc) Regular review

8. Coordinate and participate in maintaining staff performance (Level 2)

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a) Technical(1) Foundational education(2) Continuing education(3) Proficiency testing(4) State licensure(5) Competency statements(6) Training programs(7) Technical and scientific reference and research

materials(8) Problem resolution

b) Administrative(1) Management(2) Budget(3) Personnel(4) Communication(5) Consultation(6) Laws and regulations(7) Records retention

9. Educate or train others (Level 3)a) Laboratory science studentsb) Healthcare personnelc) Co-workers

10. Maintain knowledge and skills through continuing education (Level 2)

B. Laboratory safety1. Describe (Level 1) and utilize (Level 2) accepted safety

precautions to prevent laboratory acquired infectionsa) Standard precautions

(1) Hand washing(2) Personal protective clothing/equipment

b) Engineering controls(1) HEPA filtration(2) Ultraviolet germicidal irradiation(3) Negative pressure room

c) Emergency action protocol d) Traininge) Health care facilities

(1) Emergency care

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(2) PPD skin test monitoring or gamma interferon testing

(3) Chest x-ray(4) Respirator fit testing(5) Treatment

f) Aseptic techniquesg) Handling and disposal of sharpsh) Use of biological safety cabinets i) Centers for Disease Control and Prevention

(CDC)/biological safety level (BSL) classification(1) Classifications of BLS requirements(2) Correlation of specific organisms and required

BSLj) Bio hazardous material discard

k) Decontamination, disinfection, sterilizationl) Emergency first aid, eye wash, showers

m) Immunizationsn) Employee health services

2. Monitor compliance (Level 2) and train others (Level 3)3. Take immediate and appropriate action when an incident

occurs (Level 2)4. Institute new guidelines and regulations (Level 2)5. Describe the procedures used to prevent aerosolization of

microbial agents (mycobacteria and other bacteria, fungi, viruses) (Level 1)a) Aseptic techniquesb) Containment proceduresc) Decontamination, disinfection, sterilizationd) Centrifuge usee) Bio hazardous materials discard

6. Describe the collection and discard of infectious waste materials (Level 1)a) Environmental Protection Agency (EPA)/state

regulationsb) Definition of infectious waste

7. Discuss hazards of chemicals in the workplacea) Safety data sheets (SDS) b) Storage, labeling and use

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(1) Physical hazards (a) Flammable(b) Oxidizer(c) Corrosive

(2) Health hazards(a) Toxicity(b) Carcinogenicity

(3) Environmental hazards8. Outline fire safety guidelines

a) Fire protocol (RACE - rescue, alarm, contain, extinguish)b) Classes of fire extinguishersc) Fire evacuation pland) Fire extinguisher protocol (PASS -pull pin, aim, squeeze,

sweep base of fire)C. Laboratory Information System (LIS)

1. Describe data entry (Level 1)a) Automatedb) Manual

2. Describe the reporting of data (Level 1)3. Discuss data retrieval to provide relevant information for

microbiology (Level 1)a) Analysisb) Integrationc) Antibiogram

4. Participate in data retrieval to provide epidemiologist/hospital infection prevention personnel with surveillance and infection prevention information (Level 2)

5. Describe the retrieval of information by clinics/providers (Level 1)a) Resultsb) Services providedc) Specimen handlingd) Education

D. Method verification and validation1. Compare methods (Level 2)2. Develop experimental design (Level 3)3. Generate sufficient quality/quantity of data (Level 2)4. Collect data, maintain records (Level 2)

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5. Analyze data (Level 2)a) Major errorsb) Minor errors

6. Make conclusions (Level 3)7. Implement changes/modifications as appropriate (Level 3)

E. Administrative tasks1. Explain the responsibilities of laboratory management (Level

1) and participate in carrying out the responsibilities (Level 2)a) Personnel

(1) Safety(2) Training(3) Proficiency(4) Competency

b) Physical facilitiesc) Communication

(1) Public health authorities(2) Infection prevention/epidemiology(3) Service providers/clinicians(4) Administration(5) Education(6) HIPAA

d) Finance(1) CPT coding(2) Cost containment

e) Reporting/consultation

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