Specimen Collection 200 QS page 1-5

Do not accept specimens with NO LABEL

What is the golden rule of specimen collection?

The quality of the specimen

The quality of any microbial study is directly proportional to what?

To maintain the viability of the pathogen with minimal or no contamination from normal flora

What is the primary goal of specimen collection regarding pathogens?

Before the antimicrobial therapy begins

When should a specimen ideally be collected in relation to antimicrobial therapy?

During the acute phase of the disease (early stage)

During which phase of a disease should a specimen ideally be collected?

2-3 days

For viral infections, during what time frame of the illness should the specimen be collected?

First week

In typhoid fever, in which week is a blood culture positive?

Second week

In typhoid fever, in which week is a stool culture typically requested for positive results?

Additional supplements

Fastidious organisms require basic nutrients along with what?

Non-Fastidious

What type of organisms require only basic nutrients to grow?

Level 1

Which order of priority level is assigned to unpreserved critical or invasive specimens like CSF and blood?

Level 2

What order of priority level is assigned to unpreserved specimens containing normal flora, such as feces and sputum?

Level 3

Which order of priority level is given to specimens requiring quantitation, such as catheter tips and urine?

Level 4

What priority level is given to preserved specimens like urine in preservatives or swabs in holding media?

Level 5

What priority level is given to specimens requiring batch processing, such as sputum for AFB culture?

Maintain specimen as near to its original state with minimal deterioration

maintains viability of specimen

Within 30 mins of collection

Ideally, within how many minutes should a general specimen be transported to the laboratory?

Not more than 10 mins

Anaerobic bacteria specimens should be transported within how many minutes?

Within 15 mins

Within how many minutes should CSF samples be transported to the laboratory?

Neisseria meningitidis and Haemophilus influenzae

Which two organisms are highlighted as temperature-sensitive in the notes?

Shigella

Which genus of bacteria is identified as pH sensitive?

Swab

Which specimen collection device is least preferred for anaerobic cultures?

1954

What year was Stuart's transport medium formulated?

Neisseria gonorrhoeae

For which pathogen was Stuart's transport medium originally formulated?

Counteract or absorb inhibitor substances present in the specimen (such as fatty acids)

What was the function of charcoal in transport media like Stuart's?

Neisseria gonorrhoeae and Bordetella pertussis

Which two organisms can be killed by fatty acids if charcoal is not used?

They can utilize glycerophosphate in the medium

Why can some gram-negative rods overgrow in Stuart's medium?

1965

What year was Amies transport medium formulated?

A balanced salt solution

In Amies transport medium, glycerophosphate was replaced with what?

Into the medium rather than the swab

In Amies transport medium, where was the charcoal incorporated?

1964

What year was Cary & Blair transport medium formulated?

8.4

What is the pH of Cary & Blair transport medium?

Campylobacter sp.

Cary & Blair medium is recommended for fecal specimens suspicious for which specific bacterium?

Stool specimens only

Buffered Glycerol Saline is designed specifically for what type of specimen?

Vibrio and Campylobacter

Buffered Glycerol Saline is toxic to which two types of enteric bacteria?

Transgrow (or JEMBEC)

Name one system specifically mentioned for transporting Neisseria gonorrhoeae.

Universal Transport Media

What media category recovers chlamydia, mycoplasmas, ureaplasmas, and viruses?

Amies transport media

Which transport medium is used for the recovery of aerobic and anaerobic bacteria?

Body fluids, bone, CSF, inner ear, corneal scrapings, foreign bodies, gastric aspirate, suprapubic aspirate of urine

Which body fluids or samples require immediate transport at room temperature?

Within 2 hours at room temperature

What is the transport recommendation time and temperature for male urethral specimens in the JEMBEC system?

Within 2 hours at room temperature

What is the transport timing and temperature for blood or bone marrow?

Rectal swab, stool culture, clean voided midstream urine

Which specimens can be held within 24 hours at 4°C (refrigerator)?

Within 2 hours

Indwelling catheter and straight catheter urine specimens should be transported within what frame at 4°C?

Immediately at 4°C

Gastric biopsy specimens must be transported under what conditions?

Dacron, Rayon, flocked nylon

According to CLSI M40 Standards, what are the three recommended synthetic swab materials?

They release significantly more specimens into transport media

Why are flocked swabs recommended over wound-fiber swabs?

Calcium alginate or wooden shafts

Which swab shaft or material types should be avoided for Neisseria, Chlamydia, and HSV?

Nitrogen or CO2 atmosphere

What type of atmospheric gas combination is present in anaerobic transport tubes to protect organisms from oxygen toxicity?

Preservatives stop bacterial multiplication (preserve true colony counts), while transport media maintain viability without supporting rapid multiplication

What is the main difference in function between preservatives and transport media?

Boric Acid

What preservative is added to urine to preserve the true colony count for 24 hours?

Urines (with Boric Acid), Respiratory Exudates, Stools/Feces, Wounds, Rectal swab (in Cary-Blair)

List two specimens that CAN be refrigerated before media inoculation.

Spinal Fluids and Other Body Fluids, Genital/Cervical for Gonococcus Isolation, Blood

List three specimens that CANNOT be refrigerated before media inoculation.

$35-37^\circ C$ (or room temperature)

If processing of spinal fluids is delayed, at what temperature should they be held?

Polyvinyl alcohol (PVA) and Buffered formalin

Which preservatives maintain the integrity of trophozoites and cysts in stool?

Sodium borate

What chemical is recommended as a preservative for shipping specimens suspected of containing mycobacteria?

0.025% sodium polyanethol sulfonate

What concentration of SPS is used as an anticoagulant in blood culture tubes?

N. gonorrhoeae, G. vaginalis, S. moniliformis, P. anaerobius

Which bacteria are sensitive to higher concentrations of SPS?

1% gelatin

What substance can be added to counteract SPS toxicity for sensitive bacteria?

Heparin

Which anticoagulant is used for viral cultures and mycobacteria?

EDTA and Citrate

According to Bailey's, which two anticoagulants should NOT be used for microbiology?

It causes a false positive result for the coagulase test

Why should citrate not be used as an anticoagulant in microbiology?

Sodium Thioglycollate

What chemical agent is added as a reducing agent to improve the recovery of anaerobic bacteria?

Provides semisolid consistency to prevent oxygenation and spillage

What role does a small amount of agar play in transport media?

Anaerobes

Specimens suspected of containing which group of organisms should NEVER be stored in the refrigerator?

$$-70^\circ C$$

Tissue specimens for long-term storage should be stored at what freezer temperature?

Up to 1 week

How long can serum for serological studies be frozen at $-20^\circ C$?

Up to 3 days

For how long can feces for C. difficile testing be stored in the refrigerator before needing $-70^\circ C$ storage?

$35^\circ C$ to $37^\circ C$

What temperature range defined as "body temperature" is used for CSF and blood storage/incubation?

Label mismatch, improper transport temperature, improper transport medium, insufficient quantity, leaking container, exceeded transport time, received in fixative, anaerobic request on normal flora site, dried specimen, questionable medical value (e.g., foley catheter tip)

Name three criteria for specimen rejection in the microbiology laboratory.

Processing them produces information of questionable medical value

Why are foley catheter tips rejected for culture?

Sputum

What specimen must be microscopically analyzed to ensure it is not saliva before processing?

Homogenization

What tissue preparation method involves grinding bone and tissue?

Centrifugation (20 minutes at 3000 rpm) or filtration

How is concentration of large volumes of sterile fluids performed?

Legionella or Mycobacteria

For which two organisms is decontamination required during specimen preparation?

10-20 seconds

Swab specimens are often vortexed in 0.5-1.0 mL of saline or broth for how long to dislodge material from fibers?

Critical Value

What is defined as a result indicating that a patient has a life-threatening infection requiring immediate notification?

No (only if it indicates a life-threatening infection)

Is every positive culture considered a critical value in microbiology?

Bacterial meningitis

What critical condition does a positive CSF Gram stain or culture indicate?

Septicemia

What critical condition does a positive blood culture indicate?

Mycobacterium tuberculosis

What pathogen in an acid-fast smear represents a critical value?

Sterile site

Recovery of Streptococcus pyogenes from what type of site is a critical value?

Neonatal sepsis

Detection of S. agalactiae or HSV from a genital site of a pregnant woman at term poses a risk of what condition?

Bordetella pertussis, Brucella spp., Legionella spp.

Name two specific pathogens whose detection is automatically a critical value.

Unsuitable

Is sputum suitable or unsuitable for anaerobic culture?

Unsuitable

Are voided urine samples suitable or unsuitable for anaerobic culture?

Suitable

Is suprapubic urine aspirate suitable or unsuitable for anaerobic culture?

Unsuitable

Are throat or nasopharyngeal swabs suitable or unsuitable for anaerobic culture?

"Sulfur granules"

What specific visual observation in sputum or actinomycosis cases indicates suitability for anaerobic culture?

Unsuitable

Are vaginal or cervical swabs suitable or unsuitable for anaerobic culture?

Suitable

Are joint (synovial) fluids suitable or unsuitable for anaerobic culture?

Suitable

Is bile suitable or unsuitable for anaerobic culture?

At the hike of fever

When is the highest concentration of microbes present in the blood?

Brain Heart Infusion (BHI) Broth

What broth medium is used in conventional blood culture systems?

7 days

How many days are conventional blood culture bottles held?

70% Isopropyl Alcohol

What is the first agent used in the 2-step skin disinfection protocol for blood cultures?

Removes surface skin oils, dead skin cells, and surface dirt (defatting)

What is the purpose of the first alcohol cleanse in blood culture collection?

2% Chlorhexidine gluconate (CHG) in 70% isopropyl alcohol

What is the preferred primary antiseptic agent for blood culture collection?

10% Povidone-iodine or tincture of iodine

What are the alternative primary antiseptics used if a patient is allergic to CHG or is an infant under 2 months?

30-60 seconds

How long should Chlorhexidine gluconate air dry after application?

2-3 sets

How many sets of blood cultures should be drawn within 24 hours?