Clinical Laboratory Studies and Interpretation of Blood Gases

Chapter 7: Clinical Laboratory Studies

Learning Objectives

  • After reading this chapter, you will be able to:
    1. Explain the three phases of laboratory testing.
    2. Describe the composition of blood.
    3. Explain the importance of specimen integrity and effects on laboratory test results.
    4. Define laboratory test sensitivity, specificity, and positive and negative predictive value.
    5. Discuss the meaning of the term reference range.
    6. Describe the clinical applications and general clinical significance of increases and decreases for each component of the complete blood count, the reticulocyte count, and erythrocyte sedimentation rate.
    7. Define leukocytosis, leukopenia, relative and absolute count, neutrophilia, neutropenia, lymphocytosis, lymphopenia, monocytosis, eosinophilia, basophilia, leukemia, anemia, and hemostasis.
    8. Describe the body's response to anemia and the potential effects on the body of uncompensated anemia.
    9. Differentiate primary, secondary, and relative polycythemia and describe the adverse effects of polycythemia on the body.
    10. Describe the clinical applications of the activated partial thromboplastin time, prothrombin time, and platelet count in assessing hemostasis.
    11. Explain the clinical application and significance of the quantitative d-dimer assay.
    12. Describe the clinical applications and general clinical significance of increases and decreases in electrolyte concentrations, glucose levels, blood urea nitrogen, and creatinine.
    13. Discuss the importance of renal and hepatic panel tests as related to the management of patients with respiratory disorders.
    14. Relate lipid panel measures to the risk for atherosclerosis and heart disease.
    15. Identify the current cardiac biomarkers used to help identify acute coronary syndrome and congestive heart failure.
    16. Describe the preanalytical phase of testing in clinical microbiology.
    17. Discuss the common methods of examination of microbiology specimens (e.g., Gram stain, acid-fast stains).
    18. Explain the purpose of a microbiology culture and antimicrobial sensitivity test.
    19. Describe the collection and transport protocols for pulmonary secretions.
    20. Discuss the importance of macroscopic and microscopic examination of sputum.
    21. List the microscopic criteria used to assess the quality of a sputum sample.
    22. Explain the significance of sputum eosinophilia.
    23. Describe the indications and method of performing a bronchoalveolar lavage.
    24. Describe the macroscopic, microscopic, and chemical significance of pleural fluid examination.
    25. Explain the purpose of histologic and cytologic examinations.
    26. List the malignant tumors responsible for producing most primary lung cancers.
    27. List the types of pulmonary samples that can be examined cytologically.
    28. Discuss the general concept of skin testing and two methods to screen for tuberculosis.
    29. Given a variety of patient presentations, identify the common laboratory tests helpful in assessing the problem.

Clinical Laboratory Overview

  • Clinical laboratory tests are used to:

    • Diagnose, treat, monitor, and prevent disease.
  • Disciplines:

    • Microbiology
    • Hematology
    • Immunology
    • Transfusion medicine
    • Clinical chemistry
    • Molecular diagnostics

Phases of Laboratory Testing

  • Preanalytical Phase:

    • Related to the selection of the appropriate assays for the patient’s condition, patient identification and preparation, and specimen selection, collection, and transport.
  • Analytical Phase:

    • The actual testing performed by laboratory scientists.
  • Postanalytical Phase:

    • Involves reporting and interpretation of results.

Blood Composition and Blood Cells

  • Composition of Blood:
    • Cells (45%):
    • White blood cells (leukocytes)
    • Red blood cells (erythrocytes)
    • Platelets (thrombocytes)
    • Plasma (55%):
    • Water and soluble substances.

Laboratory Test Parameters

  • Usefulness:

    • Predictive Value:
    • Sensitivity:
      • Frequency of positive test results in patients with disease.
    • Specificity:
      • Frequency of negative test results in patients without disease.
  • Definitions of Test Results:

    • A True Positive (TP):
    • A test is positive in a patient with the disease.
    • A False Negative (FN):
    • A test is negative in a patient with the disease.
    • A True Negative (TN):
    • A negative test in a disease-free patient.
    • A False Positive (FP):
    • A positive test occurs in a disease-free patient.

Hematology Tests

  • Two Main Categories:
    1. General Hematology Tests:
    • For evaluating normal and abnormal blood cells.
    1. Coagulation Studies:
    • For evaluating blood clotting.

Complete Blood Count (CBC)

  • Determines the number of circulating red and white blood cells.
  • Determines the number and type of WBCs.
  • Red blood cells are evaluated for size and the amount of hemoglobin present.

White Blood Cell (WBC) Count

  • Total number of white blood cells in a known volume of blood.
  • Distribution of white blood cell types:
    • Neutrophils.
    • Eosinophils.
    • Lymphocytes.
    • Basophils.
    • Monocytes.
  • In healthy individuals: Neutrophils and lymphocytes comprise the majority of the WBC count.

The Neutrophil

  • Normally makes up 40% to 70% of the total white blood cell count.
  • Produced in the bone marrow, where it matures and is called into action.
  • Once a neutrophil leaves the bone marrow:
    • Enters circulating blood.
    • Marginates through the wall of the blood vessel and into surrounding tissues.

Other White Blood Cells

  • Eosinophils:

    • Make up 0% to 6% of the WBCs.
    • Play a role in allergic reactions.
  • Basophils:

    • Make up 0% to 1% of the WBCs.
    • Also play a role in allergic reactions.
  • Lymphocytes:

    • Make up 20% to 45% of circulating WBCs.
    • Useful in the fight against viral, fungal, and tuberculosis infections.
    • Comprised of B cells and T cells (50-85% T cells; B cells and NK cells each approximately 5-20%).
  • Monocytes:

    • Make up 2% to 10% of circulating WBCs.
    • The largest type of WBC.
    • In the lung, monocytes convert to macrophages and play a key role in clearing the lungs of inhaled dust through phagocytosis.

Nonmalignant White Blood Cell Abnormalities

  • Leukocytosis:
    • An abnormal increase in circulating WBCs.
  • Neutrophilia:
    • Increase of leukocytes due to neutrophils.
  • Leukopenia:
    • Abnormal decrease in circulating WBCs.
    • Neutropenia:
    • Drop in WBC caused by a decrease in neutrophils.
  • Eosinophilia:
    • Occurs with allergic reactions and parasitic infestations.
  • Basophilia:
    • Occurs with similar disorders that cause eosinophilia.
  • Lymphocytosis:
    • Occurs with viral infections, especially mononucleosis.
  • Lymphocytopenia:
    • A decrease in lymphocytes seen in immune deficiency states, including HIV infection.
  • Monocytosis:
    • Seen in chronic infections such as tuberculosis, and may occur in inflammatory conditions and autoimmune states.

Malignant White Blood Cell Abnormalities

  • Leukemias:
    • Involve the bone marrow and peripheral blood; characterized by accumulation of blasts, the most immature stage of a cell type.
  • Lymphomas:
    • Confined primarily to lymphoid tissue.

Red Blood Cells

  • RBCs are produced in bone marrow and have a life span of 120 days.
  • Main component of RBCs: Hemoglobin:
    • Primary function is to carry oxygen to tissues.
    • Important in maintaining acid-base balance (by acting as a buffer and transporting CO₂ from tissues to lungs).
Red Blood Cell Abnormalities
  • Anemia:
    • A low RBC count.
  • Three RBC Indices:
    • Mean Cell Volume (MCV)
    • Mean Cell Hemoglobin (MCH)
    • Mean Cell Hemoglobin Concentration (MCHC)
  • When RBCs lack adequate hemoglobin, they are described as hypochromic.
  • Polycythemia:
    • High RBC count.

Reticulocyte Count

  • Represents the final stage of erythrocyte development before the RBC is fully mature.
  • Characteristics of immature RBCs:
    • Lack a nucleus; slightly larger; have not yet assumed a biconcave shape.

Hemoglobin

  • A protein that carries oxygen to tissues and is the major component of RBCs.
  • It also plays a role in maintaining acid-base balance.
  • Composition: Each hemoglobin molecule consists of four heme groups (each with an iron molecule capable of binding oxygen) and four globin chains.

Platelet Count

  • Blood platelets (thrombocytes) are the smallest cells in peripheral blood.
  • Abnormally low platelet count is termed thrombocytopenia.
  • The lower the platelet count, the higher the risk of bleeding.
  • Important for RTs to check platelet count before performing an arterial puncture.

Erythrocyte Sedimentation Rate (ESR)

  • An ordered test to monitor inflammatory diseases.
  • An increase in various plasma proteins reduces the negative charge on the surface of RBCs, leading to quicker aggregation and stacking of cells.

Coagulation Screening Tests

  • Hemostasis:
    • Three screening tests:
    • Platelet count
    • Activated Partial Thromboplastin Time (APTT)
    • Prothrombin Time/International Normalized Ratio (PT/INR)
  • Used to monitor anticoagulation therapy (e.g., patients on heparin and Coumadin).
  • D-dimer:
    • Produced as a result of the breakdown of fibrin clots that form in the vasculature.

Chemistry

  • Measures biochemical compounds and external substances administered to patients.
  • Basic Metabolic Panel Includes:
    • Electrolytes
    • Fasting glucose level
    • Two renal function tests:
    • Blood urea nitrogen (BUN)
    • Creatinine

Electrolytes

  • Sodium (Na+):
    • Primary cation.
    • Normal values: 135 to 145 mEq/L.
    • Regulated by kidneys;
    • Hypernatremia: Occurs from water loss.
    • Hyponatremia: Caused by excessive water intake or sodium loss.
  • Potassium (K+):
    • Primary intracellular cation.
    • Normal values: 3.5 to 5.0 mEq/L.
    • Abnormal potassium levels can lead to cardiac dysfunction.
  • Chloride (Cl-):
    • Most common extracellular anion.
    • Normal values: 98 to 107 mEq/L.
    • Hypochloremia: Occurs with severe vomiting, chronic metabolic alkalosis.
    • Hyperchloremia: Occurs with certain kidney diseases and prolonged diarrhea.

Total CO₂

  • Plays a major role in acid-base balance.
  • Elevation occurs with metabolic alkalosis;
    • Decrease occurs with metabolic acidosis.
    • COPD patients may have elevated total CO₂ due to chronic CO₂ retention.

Glucose

  • Major carbohydrate in the blood.
  • Common fasting blood sugar levels: 70 to 99 mg/dL.
  • Hyperglycemia and Hypoglycemia.

Renal Panel

  • BUN and Creatinine:
    • Common tests to check on renal function; neither test is sensitive to early kidney disease; heart failure also elevates BUN.
    • Glomerular Filtration Rate (GFR): Used to measure kidney function and assess kidney disease stage.

Hepatic Panel

  • Measures proteins, including albumin and globulin, and liver-associated enzymes:
    • Alkaline phosphatase (ALP)
    • Alanine aminotransferase (ALT)
    • Aspartate aminotransferase (AST)
    • Bilirubin: A byproduct of the spleen's normal breakdown of hemoglobin.

Lipid Panel

  • Measures lipids including total cholesterol:
    • High-Density Lipoproteins (HDLs): < 40 mg/dL; referred to as good cholesterol because it removes excess cholesterol from circulation.
    • Low-Density Lipoproteins (LDLs): < 130 mg/dL; referred to as bad cholesterol as it deposits cholesterol on blood vessel walls, potentially leading to atherosclerosis.
    • Triglycerides: 30 to 149 mg/dL; the main storage form of fat in humans.

Cardiac Biomarkers

  • Chemicals that appear in the blood due to ischemic myocardial damage or stress:
    • Creatine kinase (CK)
    • CK-MB
    • Myoglobin
    • Troponin
    • B-type natriuretic peptide (BNP) used to help diagnose congestive heart failure.

Microbiology

  • Clinical microbiology includes:
    • Routine bacteriology
    • Mycology
    • Parasitology
    • Virology
    • Mycobacteriology

Specimen Sampling

  • Preanalytical phase involves specimen selection, collection, and transport; microscopic examination; culture and sensitivity; and examination of pulmonary secretions including bronchoalveolar specimens (expectoration and induction).
Sputum Collection
  • Expectorated Sputum: Preferred specimen for pneumonia diagnosis.
  • Sputum Induction: Used hypertonic saline.
  • Microscopic Examination: Assesses quality and provides information about microorganisms.

Bronchoalveolar Lavage (BAL)

  • Performed during bronchoscopy, involves injecting large fluid volumes into the lungs, collecting after it mixes with cells.
  • Used to diagnose interstitial lung disease and identify pneumonia causes.

Pleural Fluid Examination

  • Normal characteristics: clear and pale, volume of 3 to 20 mL, few WBCs, no RBCs.

  • Diseases or conditions diagnosed via:

    • Exudate
    • Transudate

Histology and Cytology

  • Biopsy of lung tissue to evaluate benign vs malignant lesions.
  • Cytology studies fluids and secretions for tumor cancer evaluations.

Skin Testing

  • Tuberculin Skin Tests:
    • PPD test
    • Alternatives include interferon-gamma release assays (IGRA), such as QuantiFERON-TB Gold In-Tube test or T-SPOT TB Test.

Chapter 8: Interpretation of Blood Gases

Learning Objectives

  • After reading this chapter, you will be able to:
    1. Identify the reference ranges for both arterial blood gas and oximetry parameters.
    2. Identify the indications for blood gas and oximetry analysis.
    3. Differentiate between invasive and noninvasive methods for measuring blood gas and oximetry parameters.
    4. Outline and explain the key procedural elements in obtaining arterial blood samples via puncture and indwelling arterial line.
    5. Apply knowledge of factors affecting hemoglobin saturation to interpret oximetry data.
    6. Apply common indices of oxygenation to assess the cause and severity of hypoxemia.
    7. Employ the Henderson-Hasselbalch equation to relate changes in PCO2 and HCO3- to pH.
    8. Describe common causes, compensatory mechanisms, and expected blood gas findings in simple respiratory and metabolic acid-base disorders.
    9. Describe common causes and expected blood gas findings in combined and mixed acid-base disorders.
    10. Identify common preanalytic, analytic, and postanalytic errors in blood gas analysis.
    11. Specify methods used to ensure valid measurement and use of blood gas data.
    12. Accurately interpret arterial blood gas and/or oximetry data.

Introduction

  • Arterial blood O2 and CO2 levels reflect lung function, and analysis helps guide treatment.
  • Mixed venous blood reflects tissue conditions; peripheral venous samples lack value.
  • Pulse oximetry reduces the need for arterial blood gases (ABGs) but does not reflect CO2 levels or acid-base status.

Indications for Blood Gas and Oximetry Analysis

  • Indicated if the patient’s symptoms, medical history, physical examination, or laboratory data suggest respiratory or acid-base abnormalities.
  • Useful in evaluating treatment effectiveness, especially during significant therapy changes affecting oxygenation, ventilation, or acid-base balance.

Sampling and Measurement

  • Invasive vs Noninvasive:
    • Invasive approaches require blood sampling via needle puncture or indwelling catheter.
    • Noninvasive methods measure blood gas parameters using sensors on the skin or evaluation of exhaled gases.

Invasive Blood Sampling

  • Prior to arterial puncture, chart review to check blood clotting ability is essential.
    • Low platelets or increased bleeding time will require longer post-puncture pressure.
    • Normal compression time: 3 to 5 mins.
  • Preferred Site: Radial artery due to accessibility, ease of stabilization, and collateral circulation from the ulnar artery.
    • Alternative sites include brachial, femoral, or dorsalis pedis arteries.
Modified Allen's Test
  • Assesses collateral circulation provided by the ulnar artery.
  • Steps to Perform Test:
    1. Patient forms a fist.
    2. Compress both radial and ulnar arteries.
    3. Patient relaxes to reveal a blanched palm.
    4. Release pressure on ulnar artery and observe the time for the hand to return to color ("pink up").
  • Adequate collateral flow occurs if pinking happens within 10 to 15 seconds.

Indwelling Catheter (A-line)

  • Indications:
    • Frequent blood sampling.
    • Management of arterial BP; found in critically ill patients.
  • Careful management needed to prevent hazards associated with setup and sampling.

Arterial Sample Handling

  • Ensure air bubbles in the sample are removed to prevent inaccurate values from CO2 and O2 equilibrating with the bubbles.
  • Samples not analyzed within 15 minutes need to be iced, but analysis must occur within 1 hour to limit cellular metabolic effects.

Noninvasive Measurements

  • Pulse Oximetry: Measures SpO2.
  • Transcutaneous Analysis: Noninvasive measurement of PtO2 and PtCO2.
  • Capnography: Measures CO2 concentrations or partial pressures in expired gases; usually about 2 to 5 mm Hg lower than PaCO2.

Assessment of Oxygenation

  • Parameters for Assessment:
    • Arterial partial pressure of oxygen (PaO2)
    • Hemoglobin content (Hb)
    • Hemoglobin saturation (SaO2, SpO2)
    • Arterial O2 content (CaO2)
Hypoxemia Assessment
  • PaO2 represents the pressure exerted by dissolved O2, reflecting the lungs' ability to transfer O2; levels gradually decrease with age.

  • Hypoxemia Categories:

    • Mild
    • Moderate
    • Severe
  • Causes of Hypoxemia:

    • V/Q mismatch
    • Shunt
    • Diffusion defects
    • True hypoventilation
    • Breathing reduced partial pressure of O2.
  • Hypoxia:

    • Refers to inadequate tissue oxygenation.
Hemoglobin Saturation
  • SaO2 reflects actual O2 bound to Hb compared with total capacity; >90% is generally considered adequate.
  • CaO2: Total amount of oxygen carried in the blood (normal: 16 to 20 mL/dL); indicates dissolved (PaO2) and bound (to Hb) oxygen.

Quantifying Oxygenation and Hypoxemia

  • PaO2:FiO2 (P:F) Ratio can be calculated to assess oxygenation.
  • Oxygenation Index (OI): OI = (FiO2 x Paw)/PaO2, where Paw = Mean Airway Pressure.
  • Alveolar Air Equation: PAO2 = F102(PB - H2O) - (Paco2 x 1.25).
Carboxyhemoglobin (HbCO)
  • Amount of CO bound to Hb (normally 0% to 1%) measured by co-oximetry.
  • CO competes with O2 for Hb binding sites having a 200 to 250 times' greater affinity than O2, decreasing O2 binding capacity.

Oxygen Dissociation Curve

  • Changes in PaCO2, body temperature, and pH cause shifts in the O2-Hb curve, affecting oxygen transport and tissue oxygenation.

Clinical Assessment of Oxygen

  • Goal: Provide adequate tissue oxygenation and evaluate:
    • Lungs’ ability to oxygenate blood utilizing ABG analysis and pulse oximetry.
    • Cardiovascular system role in blood distribution through physical assessment.

Interpretation of Blood Gas Measurements

  • Arterial and mixed venous samples are useful to evaluate:
    • Oxygen status by examining PaO2, SaO2, CO2, PvO2.
    • Acid-base balance by examining pH, PaCO2, HCO3-, BE.
    • Adequacy of ventilation by assessing PaCO2 levels.
Normal Arterial Blood Gas Values
  • PaO2: 80 to 100 mm Hg
  • SaO2: >95%
  • CO2: 16 to 20 mL/dL blood
  • pH: 7.35 to 7.45
  • PaCO2: 35 to 45 mm Hg
  • HCO3: 22 to 26 mEq/L
  • BE: 0±2
  • P(A-a)O₂: 10 to 15 mm Hg on room air
  • Mixed venous oxygen (PvO2): 38 to 42 mm Hg.

Assessment of Acid-Base Balance

  • pH: Reflects balance between blood acids and bases (normal: 7.35 to 7.45); pH
Partial Pressure of Carbon Dioxide (PaCO₂)
  • An important respiratory component and reliable indicator of ventilation effectiveness.
  • Hyperventilation: PaCO₂ <35 mm Hg causing respiratory alkalemia;
  • Hypoventilation: PaCO₂ >45 mm Hg leading to respiratory acidemia.
Plasma Bicarbonate (HCO3-)
  • Reflects the renal system's capacity to manage acids.
  • HCO3 <22 mEq/L indicates metabolic acidemia;
  • HCO3 >26 mEq/L indicates metabolic alkalemia.

Simple Acid-Base Disorders

  • Respiratory Acidosis:

    • Caused by decreased alveolar ventilation, resulting in rising PaCO₂, leading to lowered pH.
    • May result from pulmonary diseases, decreased drive to breathe, and diminished breathing abilities.
    • Compensation occurs via renal retention of HCO3-.
  • For acute respiratory acidosis:

    • Each 10 mm Hg increase in PaCO₂ results in a 1 mmol/L increase in plasma HCO3-.
  • Respiratory Alkalosis:

    • Caused by excess alveolar ventilation, leading to decreased PaCO₂ and a rise in pH.
    • Compensation occurs via renal loss of HCO3-.
  • Metabolic Acidosis:

    • Defined by plasma HCO3- or BE falling below normal, often resulting from decreased buffer production or increased acid production.
    • Compensation is achieved rapidly via hyperventilation.
    • Lack of compensation suggests concurrent respiratory defects.
  • Simple Metabolic Alkalosis:

    • Elevated plasma HCO3- or BE.
    • Compensation through hypoventilation; comatose patients may exhibit significant response.

Mixed Acid-Base Disturbances

  • Occur when two simple acid-base disorders are present simultaneously.

  • May be difficult to identify when HCO3- and PaCO₂ move in opposite directions, complicating understanding of compensation levels.

  • Indicators of Mixed Disturbances:

    • Significant changes in pH, PaCO₂, and HCO3-.
    • Especially in critically ill patients, prognosis is often poor.

Ensuring Valid Measurements

  • Avoiding errors is essential for accuracy in blood gas data collection.
    • Preanalytic, analytic, and postanalytic errors must to be mitigated.
    • Employ internal and external validity checks to ensure reliability.