Specimen Collection: BLOOD Flashcards by Kyra Catedral

Types of specimen

Feces
Anaerobic sample
Blood
Urine
Tissue biopsy
Sputum
Swab

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Test selection

Microscopy
Culture
Serologic
Genetic

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This is done to identify antimicrobial drugs that will be best used to control or kill the organisms.

All the information gathered through the tests will be utilized by the clinician to come up with the ideal treatment regimen for the patient.

antimicrobial susceptibility testing

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GENERAL SPECIMEN COLLECTION GUIDELINES

Obtain specimen before treatment;

Collect material from the appropriate site and avoid contamination.

Obtain material during the acute stage of the illness and ensure correct timing of specimen collection;

Collect using sterile collection method;

Collect sufficient quantity.

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LABEL:

Patient’s full name

Identification number of the patient

the type of specimen

date and time of collection

Collector’s signature

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Incorrect Labelling

• Label too high (on cap)
• Label too low (started on bottom)
• Labels exposed to adverse conditions: heat, cold, moisture
• Two labels should not share the same tube
• Double label cover epic label completely
• Twisted label
• Label upside down - Accession number belongs under cap
• Folded/ bubbled label
• Wrong color tube for test requested

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SPEICMEN TRANSPORT GUIDELINES

All specimen must be sent to the laboratory on the day of collection with as little delay as possible.

Bacteria are vulnerable to delay in processing

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DELAY IN TRANSPORT AND PROCESSING

Refrigeration
Ex: URINE ->

refrigerate (24 hrs)

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DELAY IN TRANSPORT AND PROCESSING

Maintain viability of bacteria and slows down all 3 processes

Transport Medium

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Transport media

Slows down: 3

• Growing process
• Reproducing process
• Dying process

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TRANSPORT OF SPECIMEN (REFERRAL TESTING)

• Volume

• Packing

• Paperworks

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TRANSPORT OF SPECIMEN (REFERRAL TESTING)
• Volume: must not exceed______
Bacteria and fungi ->_______

40 mL

tubed solid media

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TRANSPORT OF SPECIMEN (REFERRAL TESTING)

• Packing: leak-proof, tightly sealed containers with________

DOH Manual on ______of Laboratory Specimen for Referral

______Dangerous Goods Shipping
Guidelines

Biohazard label

Packaging & Transport

IATA’s

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TRANSPORT OF SPECIMEN (REFERRAL TESTING)

Paperworks: ______that are enclosed in the container, identified and marked properly

itemized content

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IATA- DOH TPS

Patient specimens or culture isolates must be______ before being shipped.

The material is placed into a primary receptacle that must be watertight.

______material is placed around the primary receptacle, and it is then placed into a secondary container that is also watertight.

The secondary package is sealed and placed into a sturdy outer container constructed of______.

Specific instructions must be followed for labeling the container as “________.” Specimens that are shipped by air require specific labeling and shipping documents.

triple packaged

Absorbent

fiberboard

Hazardous Material

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Triple Package System

Watertight Primary Receptacle
Glass, Metal, or Plastic*

Watertight Secondary Packaging

List of Contents

Rigid Outer Packaging

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made of glass tubes/metal tubes/ mostly plastic (safer)

there should be an absorbent packing material (especially for liquids) that should keep the tubes from shaking around

Inner Container/Primary Receptacle

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Placed in a second packaging that would contain the Itemized List of Contents

Inserted into a rigid cardboard that has an Infectious Substance Label, Shipping name, shipper’s name and Identification, data or information, UN number/UN package certification mark

Watertight Secondary Packaging

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SHIPPING LABELS

WHAT’S PRESENT:

Information from the sender
Information for the receiver
Package and content
Laboratory & contact number
UN Number is present
‘Infectious substance’ sticker

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Infectious substance’ sticker

If the infectious substance is_______, UN2814 is the number

If the infectious substance contains a biologically______ organism, it belongs to Biological Substance Category B, UN3373

known or identified

unknown

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Respiratory
Gastrointestinal
Blood Culture
Urine
Fungal
Mycobacterial
Chlamydia

1 hr

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Anaerobic DT

30mins

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Wound, skin & soft tissue DT

30mins

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CSE and Other fluids DT

Immediately

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Certain bacteira, such as _____ and _____ are very susceptible to drying and changes in temperature

Neissera and Haemophilus

CRITERIA FOR SPECIMEN REJECTION

1. Unlabelled specimen 2. Incorrectly labelled specimens (Mismatch Specimen and Form) 3. Specimen sent in wrong container; Inappropriate container; Wrong preservative for test 4. Broken Leaking container; Spilt sample 5. Insufficient sample volume (Volume not to the mark) e.g Citrated blood 6. Specimen received in syringe 7. Contaminated sample or Contaminated request form

REJECTED SPECIMEN: ANAEROBIC CULTURE

1. Gastric washings 2. Voided urine 3. Stool 4. Oropharyngeal samples 5. Sputum 6. Swabs of ileostomy or colostomy sites 7. Superficial skin specimen

UNIVERSAL PRECAUTIONS General rule:

Treat any specimen as infectious

Appropriate barriers (PPE)

Gloves Lab coat Masks Goggles Impermeable gowns or aprons

CLINICAL SPECIMEN

• Blood • respiratory tract (upper, lower) • Urine • Cerebrospinal fluid • Gastrointestinal Tract • Genital Tract • Wound & Abscess

Blood culture purpose

Confirm the infectious etiology Identify the etiologic agent Guide antimicrobial therapy

BLOOD CULTURE -> determines (3) • Aims to grow pathogenic microorganisms for diagnosis of above-mentioned diseases

bacteremia, fungemia & septicemia

FACTORS THAT AFFECT RECOVERY OF ETIOLOGIC AGENT IN THE BLOOD

Specimen Collection Culture Medium Incubation Requirement

Volume of blood is critical; yield affects isolation of the infectious agent - collect more than_____, minimum is_____ for adults (2 containers)

10mL 20mL

Temperature for incubation

35-37C

Frequent blood culture contaminants but also true pathogens:

• Coagulase-negative staphylococci • viridans Streptococci • Corynebacteria A • Bacillus species • Propionibacterium species

VOLUME OF BLOOD • 1 set=

2 tubes Aerobic & anaerobic

Adult One set in one arm, the other in another arm, last set is from the leg vein The more blood volume to be examined, the greater chances for you to be able to isolate organisms

20-60 mL (adult)

(pediatric patient) (neonates) (1 month - 2 years old) (children below 10 years old)

5-10 mL 1-2 mL 2-3 mL 3-5 mL

NUMBER OF BLOOD CULTURE SETS Collect______ sets of bottles (aerobic + anaerobic) for each septic episode If culture is negative after _____ incubation and patient is still potentially septic without an identified source Collect______additional sets of bottles (aerobic + anaerobic) If culture is negative after 24 h incubation…….

2 to 3 24-48 h 2 to 3 Repeat protocol if necessary Prolong incubation Investigate non-microbial etiology

= watch out for the times where the patient's temperature would spike up, this is a telltale sign that the organisms or are in their generation time period

Septic episode

= it could be viral. Viral organisms do not respond very well on culture media used for bacteria. They do not just grow on any environment.

Investigate non-microbial etiology

= sometimes, there are organisms that would be present after ***21 days of incubation;*** if not, call and inform the physician, and ask if they could repeat process for the third time around (2-3 sets per septic episode)

Prolong incubation

If there's one tube that turns out to be positive, what should we do? • If the pathogenic organism identified in that bottle is Listeria, Staphylococcus, Brucella, Haemophilus or Enterobacteriaceae...

This is a probable bloodstream infection

If there's one tube that turns out to be positive, what should we do? If viridans streptococci or coagulase-negative staphylococci and consistent with clinical setting (e-g-Indwelling catheter, prosthetic heart valve, immunocompromised patient)….

Probable bloodstream infection

If there's one tube that turns out to be positive, what should we do? If normal skin flora is positive ONLY for Propionibacterium, corynebacterium, Bacillus, coagulase-negative Staphylococci….

Probable contamination

What to do after if…. This is a probable bloodstream infection Probable bloodstream infection Probable contamination

- Repeat collection / repeat procedure - Consider non-infecitous etiology - Investigate viral etiology or non-culturable microorganism

SAMPLES FOR REJECTION

• Clotted specimens • Specimens collected using only alcohol as antiseptic • For adults: less than 20 mL of blood sample

BLOOD TO BROTH RATIO • Recommended:____ • dilute the antibacterial & inhibitory substances in the blood • Commercial Continuously Monitoring Blood Culture System may use smaller blood to broth ratio of less than____

1:10 ratio 1:5

CULTURE MEDIA

Glucose broth Bile broth Trypticase soy broth Brain heart infusion broth Thioglycollate broth Columbia or Brucella broth

o Ideal for endocarditis

Glucose broth

Ideal for samples for patients with enteric fever

Bile broth

NOT for samples that may contain Neisseria and Streptococcus pneumoniae Use BHI instead

Trypticase Soy Broth

Multipurpose broth For anaerobes

Brain Heart Infusion broth Thioglycollate broth

Anticoagulant, anti-phagocytic and anti-complementary action Neutralizes bactericidal effect of serum Inactivates certain antimicrobial agents

Sodium Polvathenol Sulfonate [SPS] (0.025 to 0.03%)

EXAMPLES OF ANTIMICROBIAL AGENTS THAT CAN BE INACTIVATED RATHER BY SPS

Gentamicin Kanamycin Streptomycin Polymyxin B

Downside to SPS • Quite toxic to certain fastidious organisms like

Neisseria, Mycoplasma, Peptostreptococcus, Streptobacillus moniliformis

Use_______- used to inactive penicillin

1-2% gelatin or penicillinase

Subculture on BAP (Blood Agar Plate) & CA (Chocolate Agar Plate): after _____hours, 3rd /5th /7th day Usually incubated on____ enhanced atmosphere

14-17 hrs. carbon dioxide

Incubation ______days is sufficient Not more than____ days; _____ = contamination

3-5 7 >5 days

7 DAYS INCUBATION IS USEFUL FOR

Fungemia Bacteremia (HACEK group, Brucella, Legionella)

Mycobacterial culture

> 4 weeks

Manual Examination of bacterial growth

Turbidity Hemolysis Gas production Pellicle formation or discrete colonies

COMMERCIALLY DEVELOPED SYSTEM

Biphasic Septicheck System Oxoid Signal Broth Displacement System

Has an agar slant attached to the broth containing bottle Allows the subculturing to be done in a closed system-we don't need to open the container to subculture the agar. Simply tilt the bottle to allow the broth to flow on the surface of the agar to allow the organism on the broth to grow on the culture media

Biphasic Septicheck System

Comes with a cylindrical signal device that's attached to the main bottle with a long needle extending to the surface of the broth. Closed system as well

Oxoid Signal Broth Displacement System

Automated Detection System

BACTEC 9240/ 9120/ 9050 BAC T/Alert TREK ESP culture system

• Alerts when the culture is positive • You can process a lot at one time Gram stain & subculture follows

Automated detection system

CONTINUOUS MONITORING SYSTEMS ADVANTAGES

• • • Decrease the laboratory work Decrease the pseudo bacteremia due to decrease in sampling and handling Increase in the speed of detection and rate of recovery

DISADVANTAGES OF CONTINUOUS MONITORING SYSTEMS

Requires an instrument Limited selection of medium Expensive

Specimen Collection: BLOOD Flashcards

(70 cards)