Specimen collection and Handling Flashcards
Specimen Collection Timing
Collect during ACUTE/EARLY PHASE OF ILLNESS (2-3 days for viral infection). Collect before administration of antibiotic, antifungal, or antiviral.
Specimen Storage
Collect and store specimen in STERILE SCREW-CAP CONTAINER except for STOOL
Swab Use - General
POOR specimen if aspirate or biopsy can be obtained
Swab Use - Appropriate
nasal/nasopharyngeal, oropharyngeal, eyes, ears, or genitals
Swab Use - Not Appropriate
NOT FOR ANAEROBES AND FUNGI
Swab Shaft - Toxic to Chlamydia and Mycoplasma
Wooden
Swab Shaft - Used usually in a calcium alginate-tipped swab
Aluminum
Swab Shaft - General purpose shaft for bacteria and viruses
Plastic
Swab Tip - General use for bacteria and viruses
Dacron
Swab Tip - Excellent collection and release of samples
Rayon
Swab Tip - Toxic to Neisseria (unless with charcoal)
Cotton
Swab Tip - Toxic to enveloped virus & Neisseria, interferes with PCR methods
Calcium alginate
Swab Design - Can trap organisms, leading to decreased recovery/sensitivity
Woven/Traditional
Swab Design - Uses Nylon fibers for enhanced uptake and release of microbes
Flocked
Enhancing Microbe Recovery
Place swab in 0.5-1.0 mL of sterile broth/saline and vortex it
Specimen Transport - Container
Leak-proof plastic bag with separation for paperwork; has BIOHAZARD label
Specimen Transport - Time Frame
Transport within 2 hours to the lab, except sterile specimens (within 15 minutes)
SARS-CoV-2 Transport Medium
3mL of viral transport medium (VTM), Amies transport medium, phosphate buffered saline, or sterile saline
Bacteria Susceptible to Air Exposure
Anaerobes
Bacteria Susceptible to Temperature Changes
Neisseria meningitidis
Bacteria Susceptible to pH Changes
Shigella
Bacteria Susceptible to Glycerol
Vibrio
Shipment - Arrangement of Container
Sample > Primary container > Absorbent > Secondary container > Dry ice/Liq. nitrogen > Outer/Tertiary container
Shipment - Category A
Substances/microbes capable of permanent disability or life-threatening
Shipment - Category B
Substances/microbe incapable of permanent disability or life-threatening on its current form
Preservation - Urine
Use BORIC ACID (bacteriostatic for 3 days)
Preservation - For Parasites
Don’t use formalin or Polyvinyl alcohol (PVA)
Preservation - Stuart/Amies Medium
Contains charcoal to neutralize fatty acid toxic to bacteria with pili
Preservation - Anticoagulants
Prevent clotting in blood, bone marrow, synovial fluid
Preservation - SPS (Sodium Polyanethol Sulfonate)
0.025-0.050% concentration, anticoagulant, anticomplement, antiphagocytic
Preservation - SPS - Function
Neutralizes aminoglycoside, neutralize with 1% gelatin
Preservation - SPS - Anaerobe Sensitivity
0.025% concentration preferred for some anaerobes and Neisseria
Preservation - Heparin
Can be used for viral culture and Mycobacterium culture, inhibits some gram-positive bacteria and yeasts
Preservation - Unused Anticoagulants
Citrate, EDTA, and others – should never be used
Preparation - Plating
Sterile specimens are plated directly into the culture media
Preparation - Centrifuge
> 1 mL of fluid samples centrifuged at 3000 g for 20 minutes
Preparation - Tissue Homogenization
Tissues may be homogenized using a tissue grinder (disadvantages: destroys fungi)
Specimen for Abscess, Lesion, Wound, Ulcer container
Swab in Amie’s or Stuart’s medium
Common Isolates Abscess, Lesion, Wound, Ulcer - Macule
Dermatophytes, T. pallidum
Common Isolates -Abscess, Lesion, Wound, Ulcer Papule
HPV, S. aureus, Pox
Common Isolates -Abscess, Lesion, Wound, Ulcer Nodule
C. diphtheriae, Sporothrix, M. marinum
Common Isolates - Abscess, Lesion, Wound, Ulcer Pustule
Candida, Herpes, N. gonorrhoeae, S. aureus, VZV
Common Isolates - Abscess, Lesion, Wound, Ulcer Bulla
C. perfringens, HSV, S. aureus, V. vulnificus
Common Isolates - Abscess, Lesion, Wound, Ulcer Ulcer
B. anthracis, H. ducreyi, T. pallidum
Blood - Adult Volume
≥ 20mL/set
Blood - Pediatric Volume
1-20mL/set
Blood - Culture Bottle
Both aerobic and anaerobic
Blood - When to Draw
During febrile episodes, 2-3 sets per 24 hours, Do not draw >4 sets in 24 hours
ARD (Antimicrobial Removal Device) - Manual
1:10 blood to broth ratio, Growth detected in pellicle/turbidity/hemolysis/bubbles
ARD - Automated
1:5 blood to broth ratio, Growth detected by BacT/Alert (5 days TAT)
Common Blood Isolates
Staphylococcus aureus, E. coli, Staphylococcus epidermidis, Enterococcus, B-hemolytic streptococci, Candida albicans, Pseudomonas aeruginosa, Klebsiella pneumoniae, Enterobacter cloacae, Proteus, Bacteroides, Clostridium
Bone Marrow - Culture Bottle
May use blood culture bottle if volume is sufficient
Common Bone Marrow Isolates
Brucella, Histoplasma
Amniotic, Peritoneal, Pericardial, Pleural Fluids container
Sterile screw cap or blood culture bottle, Transport within 15 minutes at room temperature
Common Fluid Isolates - Pericardial
Coxsackie, Adenovirus, Influenza