3: Specimen Collection and Handling

Without tables for Selection of Appropriate Culture Medium, Incubation Conditions, and Incubation Time

Specimen Collection

* An essential procedure before performing any laboratory test
* Involves fundamental skills imperative for correct diagnosis

False

T/F: Specimen collection only focuses on proper collection of specimens

acute phase prior to administration of antibiotics

**Principles in Specimen Collection**

Know the right time to collect the specimen. In bacteriology, the specimen must be collected at what phase of the infection?

laboratory request

**Principles in Specimen Collection**

Specimens must be properly labeled and accompanied by a (?) signed by the attending physician

specific instructions

**Principles in Specimen Collection**

Provide (?) on how to collect specimens to patients and other health care personnel

False

**Principles in Specimen Collection**

T/F: Avoid contamination; Normal flora are not considered possible contaminants

leak-proof

**Principles in Specimen Collection**

Use sterile containers that are clean and (?)

True

**Principles in Specimen Collection**

T/F: use sterile containers except in stool collection

Swab

**Principles in Specimen Collection**

How do you properly collect aerobic bacteria?

Aspirates

* placed in an anaerobic transport system

**Principles in Specimen Collection**

How do you properly collect anaerobic bacteria?

anatomic site

**Principles in Specimen Collection**

For lesions, wound, and abscesses, identifying first the exact (?) of collection and cleanse it properly

advancing margin

**Principles in Specimen Collection**

Collect specimen at the (?)

Needle aspiration

* place into an appropriate tube or transport vial

**Principles in Specimen Collection**

For deep wound, collect by (?)

Swabbing technique

**Principles in Specimen Collection**

For superficial wounds, collect by (?)

* upper respiratory tract
* nasopharyngeal
* external ear
* eye
* genital tract

**Principles in Specimen Collection:** Swabs

Swabs are appropriately used for collecting specimens in which parts of the body?

Throat cultures

**Principles in Specimen Collection:** Swabs

Swabs for the Upper Respiratory Tract

* what type of culture if isolationg Group A Streptococcus

Nasopharyngeal cultures

**Principles in Specimen Collection:** Swabs

Swabs for the Upper Respiratory Tract

* what type of culture if diagnosing pertussis, middle ear infections, carrier state of *S. aureus*

* gonococcal urethritis
* vaginosis
* vaginitis

**Principles in Specimen Collection:** Swabs

Swabs for the Urogenital

* what are being diagnosed

False

**Principles in Specimen Collection:** Swabs

T/F: Always collect once when using swabbing technique

* dacron
* calcium
* alginate
* rayon

**Principles in Specimen Collection:** Swabs

Types of swabs used in specimen collection

* which are used for bacterial culture

* cotton
* dacron

**Principles in Specimen Collection:** Swabs

Types of swabs used in specimen collection

* which are used for viral culture

* flexible swab

**Principles in Specimen Collection:** Swabs

Types of swabs used in specimen collection

* which are used for nasopharyngeal swab

Urine

**Principles in Specimen Collection**

* collected for the diagnosis of UTI

1. Cystitis
2. urethritis
3. pyelonephritis
4. pyelitis
5. glomerulonephritis

**Principles in Specimen Collection:** Urine

Name the 5 types of UTI

100,000 CFU/mL and above

**Principles in Specimen Collection:** Urine

As a general rule, a colony count of (?) is indicative of UTI

Clean catch midstream

**Principles in Specimen Collection:** Urine

Preferred urine sample collection method for culture

False

**Principles in Specimen Collection:** Urine

T/F: Random voided urine and Foley catheters are acceptable for urine specimen collection

1. First morning
2. Catheterized
3. Plastic bag (disposable pediatric urine)
4. Suprapubic aspiration
5. Collection during cytoscopy

**Principles in Specimen Collection:** Urine

Name 5 other methods of collecting urine sample

urethra meatus

**Principles in Specimen Collection:** Urine

Procedure

* Ask male patient to clean around (?) to reduce risk of contamination

uncircumcised

**Principles in Specimen Collection:** Urine

Procedure

* (?) men should retract foreskin

labia

**Principles in Specimen Collection:** Urine

Procedure

* Women should part the (?) and clean from front to back

15-30

**Principles in Specimen Collection:** Urine

Procedure

* Ask patient to pass (?) mL of urine into toilet to wash away any bacteria colonising distal urethra

middle

**Principles in Specimen Collection:** Urine

Procedure

* Ask patient to collect the (?) part of the stream in a wide neck sterile container and void remaining urine in toilet

pot

**Principles in Specimen Collection:** Urine

Procedure

* Complete specimen request form and label on the specimen (?), document in patient’s notes that the specimen was taken

Sputum

**Principles in Specimen Collection**

Needed for the dioagnosis of pulmonary tuberculosis (PTB) and other lower respiratory tract infections (bacterial pneumonia)

gargle

**Principles in Specimen Collection:** Sputum

Procedure

* (?) water or rinse mouth prior to collection

deep cough

**Principles in Specimen Collection:** Sputum

Procedure

* expectorate sputum by performing a (?)

saline mist

**Principles in Specimen Collection:** Sputum

If patient has difficulty expectorating sputum, collection can be induced by inhaling (?)

5-10

**Principles in Specimen Collection:** Sputum

how many mL are needed for the proper collection of essential sputum specimen

1. expectorated
2. induced
3. gastric contents (children)

**Principles in Specimen Collection:** Sputum

3 types of collecting sputum sample

2 specimens in separate days

**Principles in Specimen Collection:** Sputum

Diagnosis of PTB

* how many specimens are needed

* transtracheal aspirate
* BAL
* bronchial washings

**Principles in Specimen Collection:** Sputum

Diagnosis of PTB

* other possible specimens for diagnosis

early morning specimen

**Principles in Specimen Collection:** Sputum

Diagnosis of other infections

* what type of specimen is required

pre-examine

**Principles in Specimen Collection:** Sputum

Prior to culture and sputum examination, (?) the submitted sputum

employ Bartlett’s grading system or Bartlett’s classification

**Principles in Specimen Collection:** Sputum

how to pre-examine sputum samples?

0

**Principles in Specimen Collection:** Sputum

Bartlett’s grading system or Bartlett’s classification

* grade for

\+1

**Principles in Specimen Collection:** Sputum

Bartlett’s grading system or Bartlett’s classification

* grade for 10-25 of neutrophils/LPF

\+2

**Principles in Specimen Collection:** Sputum

Bartlett’s grading system or Bartlett’s classification

* grade for >25 of neutrophils/LPF

\+1

**Principles in Specimen Collection:** Sputum

Bartlett’s grading system or Bartlett’s classification

* grade for presence of mucus

\-1

**Principles in Specimen Collection:** Sputum

Bartlett’s grading system or Bartlett’s classification

* grade for 10-25 of epithelial cells/LPF

\-2

**Principles in Specimen Collection:** Sputum

Bartlett’s grading system or Bartlett’s classification

* grade for >25 of neutrophils/LPF

0

**Principles in Specimen Collection:** Sputum

Bartlett’s grading system or Bartlett’s classification

* score indicating lack of inflammation or presence of saliva

True

**Principles in Specimen Collection:** Sputum

Bartlett’s grading system or Bartlett’s classification

* T/F: the presence of >10 epithelial cells can be accepted for culture UNLESS >25 neutrophils are present

Unsatisfactory

**Principles in Specimen Collection:** Sputum

Bartlett’s grading system or Bartlett’s classification

* >10 squamous epithelial cells (SEC) /100x total magnification

Unsatisfactory

**Principles in Specimen Collection:** Sputum

Bartlett’s grading system or Bartlett’s classification

*

Unsatisfactory

**Principles in Specimen Collection:** Sputum

Bartlett’s grading system or Bartlett’s classification

* >10 SEC and

Unsatisfactory

**Principles in Specimen Collection:** Sputum

Bartlett’s grading system or Bartlett’s classification

* >25 SEC /OIF

gram-stain

**Principles in Specimen Collection:** Sputum

staining requirements for sputum samples

Stool

**Principles in Specimen Collection**

specimen of choice for the diagnosis of gastrointestinal pathogens

3

**Principles in Specimen Collection:** Stool

collect stool specimen for (?) consecutive days

2

**Principles in Specimen Collection:** Stool

another alternative in stool preservation is refrigeration up to (?) hours only

Rectal swab

**Principles in Specimen Collection:** Stool

(?) may be used if stool is present on thw swab during sample collection

Blood

**Principles in Specimen Collection**

needed for the diagnosis of the following:

* fever of unknown origin
* septicemia
* bactermiea
* typhoid fever
* subacture bacterial endocarditis (SBE

* before and after the fever spike
* 2-3 cultures placed an hour apart

**Principles in Specimen Collection:** Blood

best time to collect blood for culture

70% alcohol

povidone iodine / chlorhexidine

**Principles in Specimen Collection:** Blood

Procedure - Venipuncture

* pior to venipuncture, the area is cleansed with (?) followed by (?)

1-2

**Principles in Specimen Collection:** Blood

Procedure - Venipuncture

* the iodine is allowed to dry for how many minutes

24

**Principles in Specimen Collection:** Blood

Procedure - Venipuncture

* blood is collected on both arms within how many hours

approx 20 mL

**Principles in Specimen Collection:** Blood

Procedure - Venipuncture

* recommended volume of blood for adults

approx 1-20 mL (1-5 mL)

**Principles in Specimen Collection:** Blood

Procedure - Venipuncture

* recommended volume of blood for pedaitrics

True

**Principles in Specimen Collection:** Blood

Procedure - Venipuncture

* T/F: aerobic and anaerobic collection tubes may be used

1:10

**Principles in Specimen Collection:** Blood

Procedure - Venipuncture

* ratio of blood and culture media

neutralizers

* antimicrobial removal device (ARD)
* penicillinase
* para-aminobenzoic acid (PABA)

**Principles in Specimen Collection:** Blood

Procedure - Venipuncture

* if a patient started w/ antibiotic therapy, use blood collection tubes with (?)

37 C

**Principles in Specimen Collection:** Blood

Procedure - Venipuncture

* blood is incubated at what temperature

manual

**Principles in Specimen Collection:** Blood

Procedure - Venipuncture

* method for blood culture utilized

* BacT/ALERT
* BD BACTEC
* VersaTREK

**Principles in Specimen Collection:** Blood

Procedure - Venipuncture

* if not manual method, name other possible automated blood culture systems used at present

broth

* BHI
* TSB
* Biphasic media
* Thioglycollate broth

**Principles in Specimen Collection:** Blood

Procedure - Venipuncture

* media of choice

* SPS (0.025-0.030%)
* heparin

**Principles in Specimen Collection:** Blood

Procedure - Venipuncture

* anticoagulant used

blood culture bottles and tubes

**Principles in Specimen Collection**

SPS yellow top is used for (?)

heparin

**Principles in Specimen Collection**

for viral culture, or if the suspected organism is *Mycobacterium spp.*, what anticoagulant is used?

body fluids

**Principles in Specimen Collection**

* Collected by needle aspiration
* Disinfect the site prior to aspiration
* Place immediately in culture media

1. abdominal
2. amniotic
3. ascitic
4. bile
5. synovial
6. pleural
7. pericardial

**Principles in Specimen Collection:** Body fluids

list the 7 body fluids

Gastric specimens

**Principles in Specimen Collection**

* use of gastric tubes
* levine
* rehfuss

Cerebrospinal fluid

**Principles in Specimen Collection:** Body fluids

* collected for the diagnosis of meningitis or meningoencephalitis

* *Neisseria meningitidis*
* *Haemophilus influenzae type b*
* *Streptococcus pneumoniae*

**Principles in Specimen Collection:** CSF

possible pathogens for meningitis or meningoencephalitis

lumbar tap (3rd to 4th lumbar vertebra)

**Principles in Specimen Collection:** CSF

manner of collection

3

**Principles in Specimen Collection:** CSF

samples obtained are placed in (?) tubes

2nd

**Principles in Specimen Collection:** CSF

which tube is given to microbiology/bacteriology

approx. 1mL

**Principles in Specimen Collection:** CSF

recommended volume for culture

True

**Principles in Specimen Collection:** CSF

T/F: csf samples should processed immediately

Tissue

**Principles in Specimen Collection**

specimens obtained through biopsy

* excisional
* incisional

Aspirates

**Safety**

These must be placed in a container with tight lid or in an anaerobic transport system

needles

**Safety**

Do not accept syringes with the (?) still attached

Specimen containers

**Safety**

(?) should also be placed in a leakproof plastic container during transport

30 minutes (up to 2 hours)

**Preservation, Storage and Transport**

specimens must be ideally transported to the laboratory within how many minutes

* add preservatives
* add anticoagulants
* place in transport/culture media

**Preservation, Storage and Transport**

if immediate transport is not possible, what can be done?

1. boric acid
2. refrigeration

**Preservation, Storage and Transport**

2 possible preservatives

boric acid

**Preservation, Storage and Transport**

preservative used for urine samples, preserving it for colony count up to 24 hrs at room temp

transport/holding media

**Preservation, Storage and Transport**

maintains viability

1. caryblair
2. stuart’s / amie’s
3. JEMBEC media
* James E. Martin Biological Environmental Chamber

**Preservation, Storage and Transport**

3 possible transport/holding media

0\.025% - 0.03% Sodium polyanethol sulfonate (LIQUID)

**Preservation, Storage and Transport**

* Antiphagocytic, anticomplementary
* May inhibit some antimicrobials (aminoglycosides)
* May inhibit certain bacteria such as
* *Neisseria, Gardnerella vaginalis, Streptobacillus moniliformis, Peptostreptococcus anaerobius*