Spx Collection Prt. 2 Flashcards
Upper Respiratory Tract
Spx collection
Throat swab
Nasopharyngeal swab / aspirate
PATHOGENS OF THE URT:
• Haemophilus influenzae
• Corynebacterium diphtheriae
• Streptococcus pyogenes
• Neisseria gonorrheae
URT
Asymptomatic carriers :
• Neisseria meningitidis
• Bordetella pertussis
• Staphylococcus aureus
• Streptococcus pneumoniae
• Moraxella catarrhalis
MEDIA for URT specimen:
BAP
CAP
Thayer Martin
Bordet Gengou blood agar
MEDIA for URT spx
S. pyogenes:
H. influenzae:
N. meningitidis:
B. pertussis:
BAP
CAP or BAP w/ Staphylococcus
CAP/Thayer Martin
Bordet-Gengou blood agar
Nasopharyngeal or Throat Swab use:
2 Sterile swabs
Dacron/ Rayon swab - Synthetic fiber swabs
We do not use what type of swabs?
calcium alginate
swabs with wooden shafts
Nasopharyngeal aspirate
Aspiration:
• mucus extractor connected an ….
Nasogastric tube
Lower Respiratory Tract Collection
• Sputum
• Endotracheal aspirate
• Bronchoalveolar lavage
• Bronchoscopy secretions
PATHOGENS OF THE LRT:
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• Mycobacterium spp.
• Mycoplasma spp
• S. pneumoniae
• S. aureus
• Enterobacteriaceae
—-Enterobacter
—-Klebsiella
—-Escherichia
• Legionella spp.
• Pseudomonas aeruginosa
Collection & Transport of SPUTUM
container:
time and consistency:
delivery time:
storage time:
• Use a dry, wide-mouth bottle (screw cap)
• Collect sample early morning (purulent)
• Transport ASAP
• may be refrigerated but examined w/in 2-3 hrs
Suitability of Sputum for Culture
Bartlett’s Sputum Classification
< 10 epithelial cells
> 25 pus cells
Bartlett’s Classification
Number of neutrophils per low-power field
Fewer than 10
10-25
Greater than 25
Mucus
Fewer than 10 – 0
10-25 — +1
Greater than 25 — +25
Mucus — +1
Bartlett’s Classification
Number of epithelial cell per low-power field
10-25
Greater than 25
10-25 — -1
Greater than 25 — -2
Bartlett’s Classification
Scores of 0 or less indicate…
lack of inflammation or presence of saliva.
PROCEDURE for SPUTUM
+ Make a direct smear (GS & AFS)
+ Do digestion & concentration
+ Culture
Sputum
What to add? And why?
digestion & concentration technique
N-acetyl-L-cysteine-NaOH (NALC-NaOH)
• dissolve fats & mucus to free bacteria
URINE
Purpose:
Diagnosis of UTI
URINE
Method of collection:
Clean - catch midstream
Suprapubic aspiration
Cystoscopy or catheterization
URINE
Collection Time:
Early morning
URINE
• Processing:
Within 1 hour after collection
PROCESSING OF URINE
• GS of uncentrifuged urine
= rapid UTI screening
GS of uncentrifuged urine
= numerous squamous cells indicates…
(vaginal / urethral contamination)
URINE PROCEDURE FOR CULTURE:
1. Inoculate sample on…
- Incubate overnight
(+) growth
(-) no growth
BAP, EMB, Mac with a 1uL loop
(count colonies)
re-incubate for 24 hrs
Computation for CFU/mL
Actual # of colonies x calibration of loop = # of CFU/mL
CFU/mL that indicates infection
> 100,000 bacterial CFU/mL
POUR PLATE METHOD:
• prepare______ dilution of urine w/ sterile H2O
• transfer_____ of solution to a Petri dish
• add_____ & mix well
• incubate for_____
• count colonies & multiply by____
• report result in bacteria/mL
1:1000
1 mL
Agar
agar
1000
CEREBROSPINAL FLUID
• Samples should be collected…
• Avoid delay because of high mortality rate & rapid proliferation of microbes is associated w/…
before treatment
meningitis
CSF PATHOGENS
•Streptococcus pneumoniae
•Haemophilus influenzae type b
•Neisseria meningitidis
Cerebrospinal Fluid
• Collection:
lumbar puncture
L3 & L4
CSF
• Volume:
Adults
0.5 - 5.0 ml
3 tubes
CSF
1st tube =
2nd tube =
3rd tube =
Chem / Sero
Microbiology (GS & culture)
Hematology
CSF Processing
Macroscopic Examination
• Volume (3-5 mL)
• Color
• Appearance
CSF processing
Normal CSF
Abnormal CSF
• Clear and colorless
• Xanthochromic, bloody, cloudy
CSF
PROCEDURE:
• Centrifuge CSF
•_____ CSF on recommended media:
• Make a smear for______
•______
(2000 pm for 20 mins)
Culture
GS and India ink
Rapid Antigen Testing (RAT)
CSF culture media
Trypticase Soy Broth / thioglycollate
BAP for Gram (+) cocci
CAP for Gram (-) cocci
EMB/Mac for Gram (-) bacilli
CSF
Transport to LABORATORY <1
Centrifuge at 2000 rpm/ 20 mins.
Supernatant:
Sediment:
Latex agglutination
Gram stain/ CAP & BAP
CSF
Transport to lab >1
Inoculate______
Incubate overnight_____
Subculture to_____
Trans-Isolate Medium
35C in CO2
CAP & BAP
For Delays In CSF Processing
• CSF for bacterial culture:
Incubate @_______ OR;
Stand @________
35° C for not > 12 hrs (6)
room T° not > 1 hour
CSF for BACTERIAL CULTURE
DO NOT REFRIGERATE!
- Some orgs. are sensitive to_____
Such as ____ and _____
low T°
N. meningitidis
H. influenzae
• CSF for viral culture:
______immediately
If held for more than 24 hrs,______ specimen at_____
Refrigerate
freeze= -70°C
STOOL
Freshly collected stool (______stage of a disease)
•_____may be used
early
Rectal swab
STOOL
•Gastric aspirate ->
•Gastric biopsy ->
AFB
Helicobacter pylori
STOOL
Amount:
Container:
Transport time:
Transport medium:
1-2g
Clean, wide mouth with lid
2 hours after collection; 24 hrs @ 4°C
Cary Blair
STOOL
PROCEDURE:
Put rectal swab in_____
_____is NOT usually done but helps in identifying possible etiologic agents
enrichment broth (Selenite broth)
GS
Stool Culture:
1St day
Inoculate the specimen and incubate it overnight at_____
2nd day
1. Check enrichment tube:_____
2. Check differential media for_____
3. With growth:_____
4. No growth:_____
(3rd day)
1. Note patterns of biochemical reactions
2. If suggestive of Salmonella, Shigella, Vibrio, DO______
* From no growth in the 2nd day: If growth occurs after doing step 3 (2nd day, DO______
* Incubate overnight and perform step 1 & 2 of day 3
35°C
+/- turbidity
LFs & NLFs
Subculture in another set of tube & plates and do biochemical tests
inoculate culture from enrichment media into EMB or Mac
serological typing
DO biochemical test
Stool Culture MEDIA
Differential (EMB, Mac)
Selective (SSA, HEA, XLD)
Enrichment (Selenite F, APW)
EXUDATES
• Wounds
• Eye discharge
• Boils
• Ear discharge
• Abscesses
• Endocervical
• Ulcers
• Urethral
• Granules
• Anorectal discharge
• Rash
TRANSUDATES
Fluids
• Synovial
• Pleural
• Pericardial
•Peritonial
• Hydrocele
COLLECTION
DISCHARGES / FLUIDS:
a. Dry wound -
b. Skin lesion -
moisten swab w/ NSS before collecting
remove crust of pustule/ vesicle cap then gently swab lesion
COLLECTION OF DISCHARGES AND FLUIDS
Note:
1. Superficial wounds ->
- Deep wound ->
collected along the edge of the wound after cleaning with sterile saline
needle aspiration
COLLECTION FOR DISCHARGES AND FLUIDS
c. Endocervical :
d. Urethra :
e. Anorectal:
use swab
use swab or scrape mucosa of anterior urethra
insert swab about 4-5 cm. into the anal canal
Collection of Irrigation, Intravenous or
Intra-arterial Catheters
= use of endoscopic procedures
EXUDATES / TRANSUDATES: Collection container
+ Aspirates:
+ Ulcerative lesions:
+ Irrigation, Intravenous
+ Intra-arterial Catheter tips
+ Swabs:
+ Fluids:
+ Corneal scraping:
Sterile vial
biopsy in sterile vial w/o preservatives
(sterile vial)
2 pc. in a sterile tube
syringe with sterile rubber stopper
direct inoculation
EXUDATES / TRANSUDATES
sport time:
TranASAP (30 mins)
EXUDATES / TRANSUDATES:
Delay in transport:
• Refrigerate
• If swabs, place in TSB or Thioglycollate
• Amies or Stuart Transport Medium
• SBA slant
GENITO-URINARY SPECIMEN
SPECIMEN
• Cervical (female)
• (Urethral (male)
• Rectal (may be paired with throat swabs)
GENITO-URINARY SPECIMEN
PURPOSE
For the determination of:
• STDs
• Vaginitis
• Urethritis
• Childbirth infections
Possible Pathogens In Anogenital Specimen:
T. pallidum
N. gonorrheae
C. trachomatis
G. vaginalis
C. albicans
HSV
N. gonorrheae:
• GS:
• CAP:
• Modified Thayer Martin:
G (-) diplococci
enriched medium + CO2
selective medium to inhibit NMB
Transport Medium For N. gonorrhoeae
@TRANSGROW
@JEMBEC SYSTEM
