Week 1 - MLS departments

What is an analyte?

Analyte = any substance capable of being identified or measured (ex: glucose, sodium, thyroid hormone) 

What makes up the core lab?

Core lab = hematology & chemistry 

•  Because it makes up the core of what a lab is (heavily automated, similar processes, lots of blood tubes, makes up the bulk of testing (where most testing is done)) 

What is considered a “stand alone” department?

Stand-alone = blood bank

What is the clinical chemistry department? Goal?

Clinical chemistry = the study of biochemical processes associated with health and disease and the techniques used to assay these processes 

• Involves the measurement of constituents in body fluids (blood, serum, and plasma) to help diagnosis, monitor, or determine the effect of treatment for disease 

• Goal = to determine abnormal values that can be attributed to where or when that disease is occurring 

• Scope/components: sample prep & QA/QC (because heavily automated), biochemistry, physiology, pharmacology, immunology, toxicology, urinalysis, analytical chemistry, technology, & microbiology 

What is the transfusion dept.? And it’s components?

Transfusion = blood regulated like a drug 

Components 

• Immunohematology = blood typing principles and science used by both transfusion depts. And blood banks 

• Transfusion services = clinical lab, typing patients, crossmatching to units, handles the issues of blood products for transfusion 

• Blood bank = segregated lab.  

  • Collecting, testing, storing, and distributing donor products 

  • Ensures safety of blood products by typing units and testing them for transmissible disease 

What is the hematology dept.? What are it’s components? What does it mainly work with?

Hematology = the study of blood cells and indices in circulating blood or bone marrow in order to determine blood-related conditions or disease 

• Mainly focused on RBC & WBC 

• Hematopoietic cells = amount of circulating blood cells is regulated by the rate of production and release from the bone marrow 

• Work with whole blood (EDTA/purple top) 

• Detect anemia, polycythemia, hemoglobin issues, leukemias 

Components:  

1. Hematology 

2. Coagulation 

3. Urinalysis & blood fluids 

What is the microbiology dept.?

Microbiology = help identify infectious disease and treat them 

• Disorders caused by microorganisms  

  • Bacteriology, mycology (fungi/yeast), virology 

• One of most common healthcare problems encountered 

  • Inpatient (including nosocomial infections – infections due to hospital stay) 

  • Outpatient 

• Define and detect pathogens 

  • While not overworking a sample (being able to tell what is part of the normal flora) 

What does the anticoagulant tube do/is?

Anticoagulant tube = prevents clotting or coagulating of blood  

• Un-clotted whole blood will produce plasma after centrifugation (EDTA/purple top) 

• When mixed with an anticoagulant, it’ll be plasma 

What does the no additive or clot activator tube do/is?

No additive or clot activator = clotted whole blood will produce serum after centrifugation 

• When whole blood is allowed to clot, it’ll become a serum 

What are common specimens and analytes for the clinical chemistry department?

Clinical chemistry =  

Common analytes = proteins, electrolytes, blood gases, lipids, vitamins, hormones, tumor markers, & enzymes 

• Look at products of specific cellular gene expression/translation 

Types of specimens: 

• Blood (whole blood, serum (more common), plasma (more common)) 

• Body fluids (urine (more common, besides blood), CSF, any other BF) 

What are common specimens and analytes for the hematology department?

Coagulation:  

• Specimen is critical 

  • Blood collection tubes 

    • Anticoagulant = will produce plasma after centrifugation 

      • Plasma retains all clotting factors (what coag. Dept. Is looking for) 

    • No additive or clot activator (will produce serum after centrifugation) 

      • Serum has no clotting factors (NOT acceptable for coag. Dept.) 

  • Sodium citrate anticoagulant (light blue) 

• 9:1 blood-to-anticoagulant ratio 

 Urinalysis 

• Cells in urine (a few cells are normal) 

• RBCs, WBCs, epis 

  • Epithelial types = squamous, transitional (from the bladder – not good in abundance), & renal tubular (from tubules – not good in abundance) 

What are common specimens and analytes for the transfusion department?

Transfusion =  

• Whole blood => RBC and plasma tested (centrifuge needed) 

What are common specimens and analytes for the microbiology department?

Specimens: all require aseptic/sterile collection technique 

• Swabs (genital, wound, throat, nasopharyngeal, etc.) 

• Urine 

• Stool 

• Tissue & bone 

• Body fluids & pus 

• Respiratory (sputum or bronchial wash/lavage. NO SALIVA) 

• Nails & hair 

• blood 

What is hemolysis?

Hemolysis = damage to RBC membranes causing release of RBC contents into plasma (hemoglobin/electrolytes) 

• Presence of hemoglobin makes serum red/pinkish, but still clear in opacity 

• NOT IDEAL FOR TESTING because it affects the reliability and validity of the results (false positive/negative)

normal color = clear or yellow

What muscle markers help diagnose cardia muscle tissue in the chemistry dept.

Creatine kinase = elevated in muscular disorders, physical activity, and cardiac trauma 

• CK-MM = mostly in skeletal muscle  

• CK-MB = mostly in smooth heart muscle -> used to help diagnose heart attacks 

• CK-BB = in brain and prostate 

Troponin = protein found in all muscle as part of actin-myosin complex 

• Specific subunits are present in specific muscle tissues 

• TnT: cardiac -> used to help diagnose heart attacks 

• TnL: cardiac -> used to help diagnose heart attacks 

• Tn0: cardia and skeletal 

What is the most common test ordered in all of ML

CBC (complete blood count) = indicates the NUMBER of circulating cells and measures helpful indices 

• Multi-parameter electronic cell counters include an automatic diluting system 

  • RBC (cell size and volume), Hgb (hemoglobin in one RBC), MCV, Hct (hematocrit = % of someone’s blood that is RNCs relative to their plasma), MCH, MCHC, RDW (provides estimate of difference in RBC sizes), WBC, Platelet, MPV 

• Ran by hematology department 

Differential – peripheral blood smear (percentage result) 

• Indicates relative percentage of each type of WBC (lymphocytes, monocytes, neutrophils, etc.) 

• Drop of venous or capillary blood smeared on slide and stained to differentiate WBCs, RBCs, and platelets 

• WBC differential is a count of 100 WBCs examined under high power, relative percentage 

• Assesses morphology of RBCs and platelet hallmarks too 

What are erythrocytes (RBCs)?

Erythrocytes (RBCs) = biconcave disks with no nucleus or mitochondria 

• Function = to transport and protect hemoglobin 

  • Contains iron to reversibly bind oxygen 

What are Leukocytes (WBCs)? (what make it)

Leukocytes (WBCs)  

1. Neutrophil (~50-70%) = major lines of defense against infection 

   • Mature form = segmented nucleus “seg” 

   • Immature form = “band” 

2. Lymphocytes (~25-45%) = responsible for much of the immune function  

   • t-cells, b-cells, & NK cells 

3. Monocytes (~3-7%) = largest circulating WBC (phagocytosis) 

4. Basophil (~0-2%) = mediate inflammation (release histamine) 

5. Eosinophil (~0-4%) = provide protection against parasites 

What are platelets?

Platelets = small, disk-shaped cell fragments, with no nucleus 

• Formed from the cytoplasm of megakaryocytes in the bone marrow 

• Necessary for hemostasis (function as “plugs” to stop blood from leaking out of damaged sub-endothelium tissue of vessels) 

What is the coagulation cascade?

Coagulation cascade = clotting factors bind platelets and begin cycle of platelet activation for aggregation 

• Then the formation and stabilization of a platelet plug (a clot) 

  • All the factors activate to form a clot 

• If any number of factors are defective, clotting issues will arise 

• Coag. can detect a hypercoagulable state (clot very quick) or bleeding issues (slow to no clot) and test which factors may be at fault 

What is measured and calculated from RBCs

RBC indices:  

Measured 

• Measure cell size and volume 

• Hemoglobin is determined by spectrophotometry 

Calculated:  

• Hematocrit = the % of someone's blood that is RBCs relative to their plasma 

• Average concentration of hemoglobin in one RBC 

• RBW (RBC distribution width) provides estimate of difference in RBC sizes 

What is Thalassemia

Thalassemia = disorders of Hgb synthesis 

• Abnormal hemoglobin results from CBC 

• Mostly diagnosed by examining peripheral blood smear (differential) for RBC morphology, CBC, or electrophoresis (chemistry dept.) 

What is HDN

HDN = hemolytic disease of the newborn  

• Postpartum evaluation (bilirubin) 

What are possible origins for kidney disease?

Kidney disease origin = lowered glomerular filtration rate (GFR) or urine output (first sign). Can be from any of the following issues... (therefore lab runs many tests to see which one is the cause) 

• Renal issue = glomerulus, tubules 

• Post-renal issue = bladder, urinary tract 

• Pre-renal (cardiac) issue = lack of adequate blood flow into kidney 

• Urinalysis results are matched with serum/plasma values to provide a complete picture of disease 

What dept. is most likely to utilize MT?

Microbiology department 

• Rapid identification 

  • Viruses 

  • Instances in need of rapid ID (Sepsis) 

  • Glow-growing bacteria 

    • Mycobacteria take 4-28 days to grow on culture media 

    • Mycobacteria tuberculosis is highly infectious, debilitating, and highly resistant to antibiotics – shows that we need a rapid ID  

• Resistance genes 

• PCR & NGS 

• Benefits = more rapid than conventional culture & clearer picture of complicated infections (ability to multiplex) 

• Drawbacks = cannot differentiate dead cells from live & cannot differentiate normal flora from pathogen & more expensive 

• Technique 

  • Multiplex PCR system = looks for numerous targets within one PCR run 

What is Antimicrobial Susceptibility Testing (AST)?

Antimicrobial Susceptibility Testing (AST) = use of chemical compounds to treat infectious diseases to destroy or inhibit disease causing organisms 

• Often needed to predict whether a specific therapy will eradicate a pathogen 

• Used as a follow up to ensure infection will not return 

• Interpretations: for an antimicrobial 

  • Susceptible = follow normal dosing & pathogen will be inhibited 

  • Intermediate = higher than normal doses may be effective 

  • Resistant = inhibition will not likely occur & not advised for use 

Why is AST/Antimicrobial therapy stewardship important?

Important because of antibiotic resistance 

• Rampant because... 

  • Natural selection of resistance genes 

    • Plasmid swapping (microbes are smart, they learn from experience and each other) 

  • Inappropriate prescription/administration 

    • Wrong antibiotic (ineffective) or not indicated (viral infection) 

  • Inappropriate use (not finishing the prescribed course) 

    • Stronger resistant bacteria survive and then grow more that are resistant to the medication 

• ANTIMICROBIAL STEWARDSHIP CAN PREVENT MULTIRESISTANT ORGANISMS 

  • Because infections of antibiotic-resistant organisms are becoming a more common cause of death 

What type of tests/techniques are used in a clinical chemistry lab?

Clinical chemistry: methodologies 

Photometry = measurement of light transmittance/absorption 

Electrochemistry = measurement of electrical signals associated with a chemical system (very common) 

• Includes potentiometry (electrolytes, pH), amperometry (electroactive analytes – glucose/glucometer), and osmometry (moles of particles per kilogram of solvent0 

Immunoassays = antibody or antigen labeled for detection 

• ELISA/EIA, FIA 

Electrophoresis = separation of charged particles in solution when electrical field is applied (protein detections usually) 

• Protein, hemoglobin 

Chromatography = separation of complex mixtures based on physical interaction between solutes and a “solid phase” (not as common in hospital lab) 

• HPLC, GC, toxic drugs, ion fragments 

 OTHER

Automation = Defined as the mechanization of chemical analyses 

• Done in order to minimize manual manipulation 

  • Removes human performance variability 

  • Rapid (better TATs (turnaround time)) 

  • Free-up personnel to perform other tasks 

  • Decreases cost over time 

Chemistry profiles = several chemistry analyses designed to assess a single physiological system or state 

• System: renal, hepatic, thyroid etc. 

• State: Metabolic (CMP, BMP), lipid, etc. 

• Most cost effective 

• Profiles/panels are all part of a test menu 

  • Reason chem. Department so busy 

Blood samples: 

• Whole blood obtained via venipuncture 

• The liquid portion of whole blood is captured after centrifugation & used for chemistry testing (liquid & cellular components portions) 

• Proper specimen collection and processing is imperative for testing 

Muscle analytes = examines enzymes and proteins released from muscle upon damage of tissues from necrosis, injury or strenuous activity 

• Creatine kinase = elevated in muscular disorders, physical activity, and cardiac trauma (three isoforms: CK-MM, MB, & BB) 

• Lactate dehydrogenase 

• Myoglobin 

• Troponin (heart attack) 

What type of tests/techniques are done in a transfusion lab?

Transfusion = immunohematology 

Immunology = the study of immunity molecules (antigens, immunoglobulins) & systems responsible for the recognition of material deemed “foreign” 

Immunohematology = combined study of immunology as it pertains to blood antigens (Ag) and antibodies (Ab) 

• Specifically, the principle of agglutination in immune reactions is used to visualize compatibility of blood products to patients 

Major blood types & compatibility:  

• ABO gene on chromosome 9 

• Antibodies produced in plasma for “foreign” antigens 

• O RBCs -> Universal donor 

• AB RBCs -> universal recipient 

Type and crossmatch = requires EDTA sample (whole blood -> RBC and plasma tested) 

• Anticoagulated blood sample is drawn and centrifuged to separate the RBCs from plasma (TYPE) 

  • Forward type -> patient RBCs are typed for Ag 

  • Reverse type -> patient plasma is typed for Ab 

• Blood component is then chosen based off of assumed compatibility, and is further crossmatched to the patient (CROSSMATCH) 

  • Ex: patient plasma is combined with RBC unit 

  • Clotting = means they are not a match 

• Massive transfusion = emergency protocol initiated in massive traumas when individuals need a large amount of blood products issued for life-saving measures 

• Maternal/fetal functions 

  • Prenatal evaluation (Rh status (weak D/partial D Rh) & RhoGAM shot) 

  • Postpartum evaluation = hemolytic disease of newborn (HDN) 

• Molecular studies in immunohematology 

  • Mainly PCR (very sensitive, high negative predictive value 

    • HLA typing

      • Tissue transplants 

    • Stem cell transplants 

    • Pathogen detection 

      • Viral (donor disease) 

      • Bacterial (collection sterility) 

  • ABO molecular tests  

    • Available, but not widely utilized 

    • Serology is far cheaper and faster 

What type of tests/techniques are used in a microbiology lab?

Identifying microorganisms:  

• Isolate and identify pathogens 

  • Bacteria – identified via plating  

    • Possible types of growth 

      • Single pathogen 

      • Multiple causative agents (multiple pathogens at once) 

      • Account for normal flora  

• Narrow, change, or discontinue antimicrobial therapy (once pathogen has been identified) 

  • Change empiric broad spectrum therapy to specific targeted treatment 

  • Decreases risk of multi-resistant organisms 

Direct smears = gram stain 

• Rapid stain 

• Purpose = assesses specimen quality (contamination), screens for microorganisms, & identifies morphology and gram reaction 

  • Purple = + 

  • Pink = -  

• Results = assist physician with initial patient management & gram + are susceptible to some antibiotics that gram - are not  

Advancements =  

• Mass spectrometry (very fasts & inexpensive), molecular techniques, & total laboratory automation 

What’s the difference between a transfusion and blood bank dept.?

Transfusion

Blood bank

Transfusion services = clinical lab, typing patients, crossmatching to units, handles the issues of blood products for transfusion (where donor samples go to)  Type and screen  ABO and Rh testing  Cross-matching donor unit with patient  Antibody screening  Antibody detection test to determine non-ABO antibodies  Panel testing  Issue blood products  RBCs, Plasma, platelets  Special testing

Blood bank = segregated lab  Collecting, testing, storing, and distributing donor products  Ensures safety of blood products by typing units and testing them for transmissible disease  Function:   Donor recruitment  Establishing database of HIV positive or “at risk” donors  Component preparation and storage (packed RBCs, Plasma, platelets)  Donor processing (major typing (ABO & Rh determination +/-), test donor for blood-borne diseases (every unit for HIV, hepatitis, Zika, West Nile, syphilis)  Product labeling (ABO & Rh, and CMV negative)