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CLINICAL LABORATORY DIAGNOSTICS OF PATHOLOGICAL PROCESSESS IN LUNGS. Marushchak Maria. Obtaining a sputum sample. Mouth should be free of foreign objects Remove food, gum, or Tobacco Remove dentures Early morning specimen is best Induce sputum if necessary - PowerPoint PPT Presentation
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CLINICAL CLINICAL LABORATORY LABORATORY
DIAGNOSTICS OF DIAGNOSTICS OF PATHOLOGICAL PATHOLOGICAL PROCESSESS IN PROCESSESS IN
LUNGSLUNGSMarushchak MariaMarushchak Maria
Obtaining a sputum Obtaining a sputum samplesample
Mouth should be free of foreign objects Mouth should be free of foreign objects Remove food, gum, or Remove food, gum, or TobaccoTobacco Remove dentures Remove dentures
Early morning specimen is best Early morning specimen is best Induce sputum if necessary Induce sputum if necessary
Nebulized hypertonic saline or Nebulized hypertonic saline or distilled water distilled water
Chest percussion Chest percussion Postural drainage Postural drainage
CoughCough into sterile specimen cup into sterile specimen cup
Special Circumstances Special Circumstances TuberculosisTuberculosis suspected suspected
Sputum collected in negative pressure room Sputum collected in negative pressure room Early morning gastric aspirate Early morning gastric aspirate Bronchoscopy with bronchial lavage Bronchoscopy with bronchial lavage
Anaerobic culture specimen Anaerobic culture specimen Transtracheal aspiration Transtracheal aspiration ThoracentesisThoracentesis Direct lung puncture Direct lung puncture
Viral Culture Specimens Viral Culture Specimens Patient gargles and expectorates with Patient gargles and expectorates with
nutrient broth nutrient broth Nasopharyngeal swab transported in viral Nasopharyngeal swab transported in viral
medium medium
Preparation of Sputum for Preparation of Sputum for Lab Lab
Fixation of sputum for cytology (prevents air drying) Fixation of sputum for cytology (prevents air drying) Patient expectorates into jar of 70% Patient expectorates into jar of 70% EthanolEthanol Spread fresh sputum on slide and spray pap fixative Spread fresh sputum on slide and spray pap fixative
Culture specimen transport to lab Culture specimen transport to lab Sputum Gram StainSputum Gram Stain assesses sample for adequacy assesses sample for adequacy Anaerobic cultures transported in air tight container Anaerobic cultures transported in air tight container
Transport to lab for immediate plating Transport to lab for immediate plating Aerobic culture specimen Aerobic culture specimen
Bring to lab as quickly as possible Bring to lab as quickly as possible Refrigerate specimen if transport delayed Refrigerate specimen if transport delayed Consider washing specimen of oral flora Consider washing specimen of oral flora
Rinse several times with saline Rinse several times with saline Discard supernatant (non-viscous Discard supernatant (non-viscous SalivaSaliva) )
TuberculosisTuberculosis culture culture May be stored at room temperature for up to 48 May be stored at room temperature for up to 48
hour hour
Diagnosis #1: Necrotizing Diagnosis #1: Necrotizing granulomas, cervical lymph granulomas, cervical lymph node, acid-fast organisms node, acid-fast organisms present. Non-necrotizing present. Non-necrotizing
granulomas, pleura, no acid-fast granulomas, pleura, no acid-fast organisms seen.organisms seen. Cervical lymph node:Cervical lymph node:
A low-power view of A low-power view of the cervical lymph the cervical lymph node shows necrotic node shows necrotic nodules surrounded by nodules surrounded by a layer of pale a layer of pale histiocytes. Giant cells histiocytes. Giant cells and non-necrotizing and non-necrotizing granulomas were granulomas were absent. An acid-fast absent. An acid-fast stain revealed beaded stain revealed beaded bacilli consistent with bacilli consistent with mycobacteria. mycobacteria.
Pleural biopsy: Pleural biopsy: Non-necrotizing Non-necrotizing granulomas with granulomas with
central, epithelioid central, epithelioid cells and cells and
surrounding surrounding lymphoid cells are lymphoid cells are
present. Giant present. Giant cells are absent. cells are absent.
No acid-fast No acid-fast organisms were organisms were
found.found.
Sputum Gross ExamSputum Gross Exam
Estimate daily volume of Estimate daily volume of SputumSputum
Small amounts Small amounts Lung AbscessLung Abscess PneumoniaPneumonia TuberculosisTuberculosis
Copious amounts (>200 cc/day) Copious amounts (>200 cc/day) BronchiectasisBronchiectasis Bronchopleural FistulaBronchopleural Fistula
Sputum Color Sputum Color
Bloody SputumBloody Sputum ( (HemoptysisHemoptysis) ) Rusty Rusty SputumSputum (Prune-juice) (Prune-juice)
Pneumococcal PneumoniaPneumococcal Pneumonia Purulent SputumPurulent Sputum (yellow, green, (yellow, green,
dirty-gray) dirty-gray) Color alone does not Color alone does not
distinguish bacterial distinguish bacterial infection infection
Sputum TurbiditySputum Turbidity
Frothy Frothy SputumSputum (air bubbles, (air bubbles, HemoglobinHemoglobin) ) Pulmonary edema Pulmonary edema
Foamy, clear material Foamy, clear material SalivaSaliva Nasal secretions Nasal secretions
Sputum Viscosity Sputum Viscosity Bloody Gelatinous Bloody Gelatinous SputumSputum (Currant-Jelly) (Currant-Jelly)
KlebsiellaKlebsiella PneumoniaPneumonia Pneumococcal PneumoniaPneumococcal Pneumonia
Stringy Mucoid Stringy Mucoid SputumSputum (may also appear frothy) (may also appear frothy) Follows Follows AsthmaAsthma exacerbation exacerbation
Cloudy, mucoid Cloudy, mucoid SputumSputum Chronic BronchitisChronic Bronchitis
Three layered appearance (stagnant, Three layered appearance (stagnant, Purulent SputumPurulent Sputum) )
BronchiectasisBronchiectasis Lung AbscessLung Abscess
Sputum with Feculent Sputum with Feculent OdorOdor
Anaerobic infection Anaerobic infection BronchiectasisBronchiectasis
Example of Example of casecase
A. An overall view shows A. An overall view shows a circumscribed, non-a circumscribed, non-encapsulated mass of encapsulated mass of mature adipose tissue mature adipose tissue traversed by small traversed by small vessels. It is covered by vessels. It is covered by normal respiratory normal respiratory epithelium, which is epithelium, which is separated from the fat separated from the fat by a loose fibrous by a loose fibrous stroma that contains a stroma that contains a large number of large number of lymphocytes and plasma lymphocytes and plasma cells. The hemoptysis cells. The hemoptysis that can occur with that can occur with these lesions is caused these lesions is caused by the pneumonia by the pneumonia secondary to the tumor secondary to the tumor and not by the tumor and not by the tumor itself. itself.
B. At higher B. At higher magnification, the magnification, the subepithelial fibrous subepithelial fibrous tissue is chronically tissue is chronically inflamed. The inflamed. The irregular dark blue irregular dark blue area is a lymphoid area is a lymphoid aggregate. The aggregate. The underlying fat and underlying fat and blood vessels blend blood vessels blend in with the fibrous in with the fibrous tissue rather than tissue rather than being sharply being sharply demarcated. demarcated.
C. The C. The subepithelial subepithelial tissue has chronic tissue has chronic inflammatory inflammatory cells cells (lymphocytes (lymphocytes (without visible (without visible cytoplasm) and cytoplasm) and plasma cells (with plasma cells (with visible visible cytoplasm)). cytoplasm)).
What is the diagnosis? What is the diagnosis? Endobronchial lipoma Endobronchial lipoma
D. At the edge D. At the edge of the lesion, of the lesion, loose loose connective connective tissue and fat tissue and fat are mixed are mixed together. together.
Sputum Sputum MicroscopyMicroscopy
Assessing Sputum Sample Assessing Sputum Sample QualityQuality
Ideal Ideal SputumSputum Sample for Culture Sample for Culture Under 10 squamous epithelial cell Under 10 squamous epithelial cell
per low power field per low power field Many Many NeutrophilNeutrophils present (>5 per s present (>5 per
high power field) high power field) Bronchial epithelial cells present Bronchial epithelial cells present Alveolar Alveolar MacrophageMacrophages may be present s may be present
Inadequate Inadequate SputumSputum Sample Sample Over 25 squamous epithelial Over 25 squamous epithelial
cells/LPM cells/LPM
Sputum Sample Sputum Sample Preparation Preparation
Pull strand or plug of Pull strand or plug of SputumSputum onto slide onto slide
Consider buffered crystal Consider buffered crystal violet to stain cells violet to stain cells
Apply cover slip Apply cover slip View under oil immersion View under oil immersion
Cytology Stains Cytology Stains No Stain No Stain
BlastomycosisBlastomycosis Cryptococcosis Cryptococcosis
Gram StainGram Stain Gram Positive BacteriaGram Positive Bacteria Candida Candida TuberculosisTuberculosis (weakly (weakly Gram PositiveGram Positive) ) NocardiaNocardia (weakly (weakly Gram PositiveGram Positive) )
Direct Fluorescent Direct Fluorescent AntibodyAntibody Staining Staining Legionella Legionella
Wright stain or Giemsa Stain Wright stain or Giemsa Stain Intracellular organisms Intracellular organisms
Special Staining Special Staining Circumstances Circumstances
Acid-fast Mycobacteria Acid-fast Mycobacteria (Tuberculosis)(Tuberculosis)
Ziehl-Neelsen Stain (Red against blue Ziehl-Neelsen Stain (Red against blue background) background)
Kinyoun stain Kinyoun stain Less reliable than Ziehl-Neelsen stain Less reliable than Ziehl-Neelsen stain Results in quickly stained sample Results in quickly stained sample
Fluorochrome dyes (auramine, Fluorochrome dyes (auramine, rhodamine) rhodamine) Higher false positive rate than Ziehl-Higher false positive rate than Ziehl-
Neelsen stain Neelsen stain Assist greatly in identifying organisms Assist greatly in identifying organisms
Fungal OrganismsFungal Organisms
PAS staining or Methenamine PAS staining or Methenamine silver staining silver staining Histoplasmosis Histoplasmosis Coccidioidomycosis Coccidioidomycosis Aspergillus Aspergillus Mucor Mucor
KOH Preparation KOH Preparation
Microscopic findings Microscopic findings Caseous masses Caseous masses Dittrich's plugs Dittrich's plugs Curschmann's spirals (Asthma) Curschmann's spirals (Asthma) Charcot-Leyden Crystals (Asthma) Charcot-Leyden Crystals (Asthma) Bronchial casts Bronchial casts Concretions Concretions Broncholith Broncholith
Calcified particles as seen in Broncholithiasis Calcified particles as seen in Broncholithiasis Lung Cancer cells Lung Cancer cells
Central bronchus tumors Central bronchus tumors May require 4 samples to detect May require 4 samples to detect
Eosinophils (>5%): identified with Wright's Stain Eosinophils (>5%): identified with Wright's Stain Allergy Allergy Asthma Asthma
Саrре diemСаrре diem