mlab 2434 clinical microbiology summer 2005 cecile sanders keri brophy l.
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MLAB 2434 – CLINICAL MICROBIOLOGY SUMMER, 2005 CECILE SANDERS & KERI BROPHY. Chapter 7 – General Concepts in Specimen Collection and Handling. Chapter 7- General Concepts in Specimen Collection and Handling (cont’d). Basic Principles of Specimen Collection

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mlab 2434 clinical microbiology summer 2005 cecile sanders keri brophy

MLAB 2434 – CLINICAL MICROBIOLOGYSUMMER, 2005CECILE SANDERS & KERI BROPHY

Chapter 7 – General Concepts in Specimen Collection and Handling

chapter 7 general concepts in specimen collection and handling cont d
Chapter 7- General Concepts in Specimen Collection and Handling (cont’d)
  • Basic Principles of Specimen Collection
    • Specimen should be taken in acute phase of infection AND before antibiotics are administered
    • Written order must specify site of culture (example: wound on left arm)
chapter 7 general concepts in specimen collection and handling cont d3
Chapter 7- General Concepts in Specimen Collection and Handling (cont’d)
  • Avoid normal flora and colonizing organisms
  • Compare test results with suspected diagnosis
chapter 7 general concepts in specimen collection and handling cont d4
Chapter 7- General Concepts in Specimen Collection and Handling (cont’d)
  • Appropriate Collection Techniques
    • Aspirates and tissues
      • Aspirates and tissues present few problems, if collected using sterile technique
      • Lesions, wounds and abscesses; cultures should be from as deep in the wound as possible
chapter 7 general concepts in specimen collection and handling cont d5
Chapter 7- General Concepts in Specimen Collection and Handling (cont’d)
  • Swabs
    • Used only as a last resort
    • Steps
      • Clean wound
      • Explore wound
      • Obtain fresh and quality culture material
    • Should be placed in a holding medium to protect pathogens without permitting multiplication during transport
chapter 7 general concepts in specimen collection and handling cont d6
Chapter 7- General Concepts in Specimen Collection and Handling (cont’d)
  • Patient Education and Preparation
    • If patient is responsible for collecting specimen, good instructions are critical
    • Urine – midstream clean catch first morning specimen
    • Sputum – collect sputum NOT spit
    • Stools – usually 3 vials and at least 4 days after barium Xrays
chapter 7 general concepts in specimen collection and handling cont d7
Chapter 7- General Concepts in Specimen Collection and Handling (cont’d)
  • Preservation, Storage, and Transport of Specimens
    • Concerns
      • Overgrowth
      • Death of microorganisms
      • Inaccurate quantitation
      • Loss of organisms from drying
      • Protection from oxygen
      • Protection from clotting
      • Safety of transporter
chapter 7 general concepts in specimen collection and handling cont d8
Chapter 7- General Concepts in Specimen Collection and Handling (cont’d)
  • Preservatives
    • Urine – boric acid
    • Stool – phosphate-buffered saline (PBS)
  • Anticoagulants
    • Needed in any specimen that might clot (blood, serum, joint fluids)
    • Sodium polyanethol sulfonate (SPS)
chapter 7 general concepts in specimen collection and handling cont d9
Chapter 7- General Concepts in Specimen Collection and Handling (cont’d)
  • Use of Holding and Transport Media
    • Swabs placed in Modified Stuart transport medium or Cary-Blair transport medium
    • Blood placed in broth culture medium
    • Unprotected specimens
      • Sputums, body fluids, tissues, catheters, medical devices, and specimens for sterility culture
      • Should be processed ASAP
chapter 7 general concepts in specimen collection and handling cont d10
Chapter 7- General Concepts in Specimen Collection and Handling (cont’d)
  • Storage of Specimens
    • Urine, viral blood specimens, catheters and swabs should be refrigerated
    • Blood and CSF should be processed ASAP
    • Specimens for fungus cultures can be kept at room temperature
    • Respiratory and stool cultures should be processed ASAP if at all possible, but refrigerated if immediate processing is not possible
chapter 7 general concepts in specimen collection and handling cont d11
Chapter 7- General Concepts in Specimen Collection and Handling (cont’d)
  • Mailing Specimens
    • Regulated by U.S. Dept. of Health and Human Services
    • Primary container must be securely closed, watertight
    • Secondary container has sufficient absorbent material to absorb contents in case of breakage
    • Final container is an approved mailing container with biohazard symbol and contact phone number for CDC
    • Page 244 in textbook
chapter 7 general concepts in specimen collection and handling cont d12
Chapter 7- General Concepts in Specimen Collection and Handling (cont’d)
  • Safety
    • Universal Precautions
    • Specimen processing should be conducted in a Class II safety cabinet (hood)
  • Labeling and Rejection of Specimens
    • Requisitions must include source, diagnosis or history, and test(s) requested
chapter 7 general concepts in specimen collection and handling cont d13
Chapter 7- General Concepts in Specimen Collection and Handling (cont’d)
  • Unacceptable Specimens
    • Labels on requisition and on specimen must match
      • Noninvasive vs. invasive specimens
    • Rejected specimens
      • Leaking
      • Syringes with needles attached
      • Stools contaminated with urine or barium
      • Anaerobic cultures on inappropriate sources
      • Unpreserved specimens over 2 hours old
chapter 7 general concepts in specimen collection and handling cont d14
Chapter 7- General Concepts in Specimen Collection and Handling (cont’d)
        • Refrigerated blood cultures
        • Dried-up specimens
        • Specimens in formalin
  • Processing of Clinical Samples for Optimal Organism Recovery
    • Prioritization (Table 7-2, p. 247)
      • Level 1- Critical/invasive
      • Level 2 – Unpreserved
      • Level 3 – Accuracy of quantitation affected
      • Level 4 – Protected/Preserved
chapter 7 general concepts in specimen collection and handling cont d15
Chapter 7- General Concepts in Specimen Collection and Handling (cont’d)
  • Gross Examination of Specimens – check specimen and requisition
  • Direct Examination Techniques
    • Direct Microscopic Examination (Table 7-3, p. 250, text)
      • Determine quality of specimen
      • Diagnose infectious disease
      • Guide routine culture interpretation
      • Dictate the need for nonroutine processing
chapter 7 general concepts in specimen collection and handling cont d16
Chapter 7- General Concepts in Specimen Collection and Handling (cont’d)
  • Smear Preparation
    • Tissues
    • Swabs
    • Aspirates and body fluids
      • Single drop smear
      • Centrifuged sediment smear
      • Layered smear
      • Cytocentrifuged smear
      • Additives
chapter 7 general concepts in specimen collection and handling cont d17
Chapter 7- General Concepts in Specimen Collection and Handling (cont’d)
  • Direct Smears NOT useful
    • Throats, nasopharyngeal swabs
    • Urine
    • Female genital tract
  • Primary Inoculation of Routine Specimens
    • Types of Culture Media
      • Nonselective vs. Selective
      • Differential
      • Enriched
      • Broths