INTRODUCTION TO THE CLINICAL LAB, SAFETY, AND LAB SKILL NOTES

International Hospital Code System- A system of codes used to alert the staff in a medical facility in emergency situations such as fires, cardiac and/or
respiratory arrest, uncontrolled individuals, and disaster team alert. There are four codes. Code Red for fire. Code Blue for respiratory or cardiac arrest. Code
Yellow for uncontrolled individual or a threatening situation. Code Green for designated team of people in the hospital should report to a designated area.
Identification system of the National Fire Protection Association - A hazard identification system. This system provides at a glance, in words, symbols, and
pictures, information on the presence of potential health, flammability, and chemical reactivity hazards of material used in the lab. This information is provided
on the labels of all containers for hazardous chemicals
Physical Hazards
1. Electrical Safety - Avoid shock, never touch electrical outlets with
wet hands. Equipment must be grounded.
2. Fire Safety - Know fire codes , escape routes, and procedures if exit
is blocked. Know where fire blanket (to smother fire) and
extinguisher are located. Avoid causes of fire: smoking, open flames,
pipes, heating elements, and spark gaps. Know National Fire Code.
Fire door: Heavy door that is found
between areas of a building
Fire extinguisher: Types A, B, C, ABC
Type A - wood, paper, rubbish, draperies, or bedding.(contains
pressurized water)
Biological Hazards
1. Use Universal Precautions which means treat all specimen (blood,
urine,body fluids etc.) and patients as though they
have diseases such as the HIV virus, bacteria, parasites, fungi, Hepatitis
etc.
Also know biohazard symbol (See below)
2.Use protection barriers etc. gloves, masks, face shields, goggles, lab
coats or
gown, depending on the situation.
3.Do not eat, drink, chew gum or smoke in lab.
4. Dispose of all materials properly. Sharp contaminated objects go in
biohazard sharps container. Biohazard material
that is not sharp go in biohazard bags Broken glass and
other sharp objects that is not contaminated should go in broken glass
receptacle , and never use broken or chipped glassware.
5. Clean spills and clean work area with 5 - 10% bleach.
pressure 120 C, 15-20 lbs. pressure psi, 15-20.
7. An autoclave should be used to sterilize. It operates by steam under
aerosols may be generated
Health Science Technology II General Safety Rules
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Never handle electrical equipment when your hands are wet.
Never handle substances containing acid, such as clinitest tablets, when
your hands are wet.
Keep all drawers and closet doors closed.
Wipe spilled liquids off floors immediately according to proper
procedures.
When a classroom accident occurs, report it to the instructor immediately.
Keep all passageways clear and uncluttered (ie. backpacks, purses, etc.)
Always carry sharp objects with the point down.
Never put your hands near your face when using chemicals and other
irritating substances.
Never deviate from a procedure or manufacturer's instructions without
the instructor's permission.
Never place flammable objects near fire or flame.
Good body mechanics should be used when lifting to prevent back
strain.
Report faulty equipment to the instructor or supervisor immediately.

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Type B - flammable liquids, i.e. alcohol, grease, paint and oil
(Contains dry powder - sodium bicarbonate or carbon dioxide)
Type C - electrical wiring, most appliances, or switchboards
Type ABC - all types of fire ( contains monoammonium phosphate
power)
Mechanical Safety - learn to use equipment properly.
Proper Body Mechanics - lift supplies properly
Radiation Safety-avoid exposure to radiation and/or wear protective
shields. Know symbol. (See below)

6. Use biological safety hood when using specimens in a manner where
Chemical Safety
1. No mouth pipetting used for body fluid and dangerous chemicals, use
pipette bulb.
2. Wear goggles when working with dangerous chemicals, and when
splashes
are likely to occur.
3. Know where eye wash and safety showers are located and know how
and when to use them.
4. Chemical spills should be carefully cleaned up according to proper
procedures for the chemical involved.
A spill clean-up kit should be located in the lab with absorbents and \
neutralizers to clean up acids, mercury, and other spills.
5. When working with chemicals in which you may encounter toxic fumes
that could cause lung irritation and damage, use a fume hood.
6. All reagent and chemicals should be properly labeled and stored.

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Do not "horseplay" in the classroom or laboratory.
Always wear gloves when handling bodily fluids and waste.
Avoid eating, drinking, smoking, gum chewing or applying makeup in
the laboratory.
Wear a laboratory jacket or coat and closed-toe shoes.
Pin long hair away from face and neck to avoid contact with chemicals,
equipment, or flames.
Avoid wearing chains, bracelets, rings, or other loose hanging jewelry.
Disinfect work area before and after laboratory procedures and at any
other time necessary.
Wash hands before and after laboratory procedures, before putting and
after removing gloves, and any other time necessary.
Avoid tasting, smelling, or breathing the dust of any chemicals.
Wear goggles when working with dangerous chemicals, and when
splashes are likely to occur.
Handle equipment with care and store it properly.
Discard any broken glassware or infectious material into proper
containers
Keep all non essential items off desk (ie. backpacks, purses etc.)

Quality Control involves:
1.Proper patient preparation - fasting or special diets
2.Proper specimen collection - properly using containers, anticoagulant tubes, and preservatives.
3.Proper maintenance of lab machinery and supplies - calibrating machines frequently etc.
4.Proper running of controls and standard with every test done to insure reagents are working, equipment is
working, and technique is done correctly.5.Correct reporting of results - supervisor should check all results before they are reported to physicians.6.Duplicate
analyses - running test twice.

Biohazard Symbol

Radiation Symbol

National Fire Protction Association Symbol

how much FeCl3 is needed? VOLUME CONNCENTRATION PROBLEMS (Vol1) (Concentration1) = (Vol2) (Concentration2) (V1)(C1) = (V2)(C2) Volumes must be in the same units! 1. To make 300 mL of 75% alcohol.06 qt 1 L= 1000 mL 29. what is the dilution titer in tube 4. 1: 50 dilution? 2.35g = 1 oz (dry) 1Kg=1000 g 1 g = 0. Convert 42. You have 150 grams of potassium iodide and want to make it all into a 5. how many grams of NaOH are needed to make a 20% solution of NaOH.Convert 150 lb. How much NaCl must be used to prepare 3500 mL of isotonic saline? 2. In a total value of 60 mL. Convert 50 F to C 3. 1 mg = 1000 g 28. and final exam MASS Standard unit of mass = gram (g) Mass does not equal weight 1 g = 1000 mg 1 kg = 2.035oz (dry) 16oz = 1lb TEMPERATURE F = 1. Convert 3 cm3 to mL VOLUME Standard unit of volume = Liter (L) Volume of a container = l x w x h 1 Kl = 1000 L 1 L = 1.14 in to m Volume: 1. .205 lb.8 (c) + 32 C = 5/9 (F .7ml = 1 liquid oz 1 ml = 1000 L 100 mL = 1 deciliter 1 ml = 1 cm3= 1 cc = 1 cubic cm 10 ml = 1 centiliter 3.37 in 1 m = 100 cm 1 yd = 36 in 1 m = 10 dm 1 yd = 36 in 1 mm = 1000 m (micrometer) 2.32) Mass: 1.14 in to cm Temprature: 1. Convert 106F to C RATIO PROPORTION PROBLEMS 3.54 cm = 1 in 1 ft = 12 in 2. How much (mL) of 95% alcohol is needed to make 250 mL of a 70% solution of alcohol? 2. LENGTH Standard unit of length = Meter (m) 1Km = 1000 m 1 m = 39. To make 5 liters of 30% solution of FeCl3. Convert 325g to oz 2. Convert 36 C to F mass = mass vol vol 1. 2:20 dilution? 3. Convert 5 feet 6 in to m 3. to g Length: 1.Lab Math Notes Units to remember for quizzes. how many mL of 100% alcohol is needed? DILUTION PROBLEMS How would you make a 1 : 10 dilution? How much saline is in a 1 : 10 dilution of blood and saline if there is 1 mL of blood? 1.0% potassium iodide solution. Convert 24 L to L 4. Convert 62. 3. Convert 3500 L to gallons 2. What volume of 5% HCl can be made from 15 mL of a 20% solution of HCl? 3. What is can make? the total volume you 4. Convert 2. Convert 22. Using the serial dilution picture. test.6 Kg to lb.4mL to L 2.

increased in infections Renal epithelial cells neg.030 (proteinuria. Urine contents are: urea. vomiting and diarrhea Bilirubin -30 sec. what should you do with it? How long may it be refrigerated before you must collect a new one? What happens to urine after it leaves the body? Urine Total Volumes Normal Range Clinical Significance (Above normal Range) (24 hour urine collection) 750 .specific gravity is fixed at a constant level of 1. diuretic drugs. 1. diabetes) Hypersthenuria -S.) 5. renal failure. chyle Odor spicy or aromatic ( foul odor can be due to infection) WBC -2 min. yellow . ammonia. Neg. starvation. hypertension.G.mucous Smokey . bacteria. (Leukocyte esterase) Neg. yellow) Orange or gold: bilirubin .1500 ml/day Polyuria (high volumes > 2. (Ictotest) Neg.(2%sulfosalylic & 3% acetic acid) Neg .dk. bacterial infection Nitrate. fat. pancreatic cancer Urine Microscopics (refer to urine sediment chart) WBC’s /HPF 0-4 due to infection Yeast /HPF neg. UTI (urinary tract infection) Black: malaria.003-1. renal disease mucus negative to 2+ increased in infections and bladder irritations Casts/LPF neg. (Cast may contain RBC’S. Ketone -40 sec.7-9. regulation of blood pH.45sec. Suprapubic tap: 3. creatinine. Random: collected at any time of day or night.lt. WBC’S. Catheterization: 2. First void: 1st urine on the day (best because it is concentrated) 7. malignant melanoma Transparency Clear Cloudy or turbid . Urine chemistries done with Multistix Reagent Strips. Clean Catch: (Does not yield consistent results. 4. liver damage Glucose -30 sec. Neg.RBC's Milky . & Clinitest) Neg. constantly below 1. leucine.WBC.500 ml/day) due to alcohol.200-1. UTI RBC’s/HPF 0-4 hypertension. Mid-Stream: middle of urine stream 6.035 Indicates the ability of the kidney to concentrate urine Urinometer or Refactometer) (Ave.) caffine. upper UTI pH -60 sec. (1.007 (hypertension. (Benedicts. sugar diabetes.1. URINE COLLECTION: TYPES OF SPECIMENS 1. diet. (Actetest) Neg.2.0 liver damage Protein -60 sec. constantly above 1. tyrosine and cholesterol are considered clinically significant. Other functions of the kidneys: excrete waste products. ( tested with 1. This is a timed test .015-1. enlarges prostate.amber *Red: indicates blood (see below) Average .G. bacteria. sugar diabetes. epithelial cells (cloudy upon standing due to crystals) Hazy . 0-5) higher in females . female cycle Squamous epithelial (occassional approx.010 (indicates very poor ability to concentrate the urine). Oliguria (low volumes) due to dehydration.Urinalysis Notes URINE is the aqueous waste product of the human body's metabolism of food which is formed in the nephrons of the kidneys. The other crystals found in urine can be considered normal . indication of renal disease. renal disease. and Hb) Crystals only crystals such as cystine. (use nitrazine pH paper) 4. hypertension. Isosthenuria . (Erhlich reagent) 0. hydroregulation.025) Hyposthenuria-S.500Ave. Conformation and back up test are in parenthesis. bacteria. and salts. kidney stones.liver damage White: pus. epithelial cells. water.0 Indicates urine freshness (old urine becomes alkaline) Blood -60 sec.fat. Urine Test Color Normal Range Clinical Significance (Above normal Range) straw.60 sec. renal disease.60 sec. maintain electrolyte balance. shock. Pediatric Collection System: If your are unable to analyze a urine specimen immediately. bacterial infection Urobilinogen. urine or vaginal yeast infection Bacteria neg. maintenance of blood pressure. diabetes. Anuria ( no urine) due to renal failure. uric acid. glucosuria). renal disease Specific gravity.

Exton's Sulfosalicylic acid method Clear or no change in urine after acid is added = negative result. Bilirubin is attracted to brain tissue and is neurotoxic. The patient will become drowsy. Jaundice is a condition which occurs when serum bilirubin levels exceed normal-skin. forms three ketones: All three ketones and can be tested for in the urine. Three major types of jaundice 1. The most common disease associated with glucosuria and hyperglycemia is diabetes mellitus (sugar diabetes). CH2O  Fats  Proteins  Fats fatty acids and glycerol  (with the aid of) acetyl Co A. If the diabetic insulin level is "out of control. They are: 1.fever 2.Benign proteinuria is transient. diabetes mellitus 3.5 g/24hr protein in the urine. associated with diabetes mellitus and hypertension. .stress 3. Hepatic-hepatitis. without ending up in the urine. Insulin acts by permitting glucose to leave the blood stream and enter inside a cell to provide the cell with nutrients for cellular growth and metabolism. RBC's→ Hemoglobin→ Heme and globin (protein) → Bilirubin When there is excess bilirubin in the blood. Bacteria in urine but no proteinuria = lower UTI. Ketosis may occur with: 1. THE LIVER (Read Textbook for functions of the Liver) FORMATION OF UROBILINOGEN AND BILIRUBIN Urobilinogen and Bilirubin are products of RBC destruction. vomiting and diarrhea. PROTEINURIA Protein in the urine is called proteinuria. False positives occur: vaginal secretions. lactose. The pH of the blood/body tissues will drop. the body will break down fats.buteric acid (2%) (20%) (78%) Ketosis in the diabetic is serious. Caused by? Speculated to be kidney blood vessels are compressed when body is upright. Bacteria in urine and proteinuria = upper UTI Types of Proteinuria Severe: 3. If a cell cannot get glucose for energy. If no glucose. Heat and acetic acid method 2. Back up tests for Proteinuria: Precipitation techniques: 1. which is the renal threshold for glucose will be spilled into the urine. Mild of minimal: less than or = to 0. benign proteinuria. Ketonemia is excess ketones in blood Ketonuria is ketones in urine. mucus. inexpensive screening tests. glucose cannot enter the cell. Prehepatic-more RBC's destroyed than normal 2. Clinitest = 0. toxemia of pregnancy 4. KETONES Ketones are substances formed from fats. The sensitivity level is 0. If urine turns cloudy after acid is added = positive result (protein present). Glucose Oxidase Method: Glucose Oxidase is an enzyme specific for glucose. stress/anxiety causes the release of epinephrine 3. Acidosis is a condition in which the pH of blood and body tissue becomes more acidic. Increase sugar/carbohydrate intake 2. cirrhosis. acetone diacetic acid B-hydroxy. blood. Hyperglycemia: Elevated blood glucose level. and respiration will be adversely affected. Benedicts = 0.120 days. About 10% of young adults have orthostatic proteinuria. the condition is called ketoacidosis or ketosis. Posthepatic-due to gall stones. infection Tests for Ketones in Blood or Urine Acetest Tablet:Contains sodium nitroprusside as the active ingredient. pregnancy 5. pus. If due to excess ketones. Renal threshold for glucose is the amount of glucose that can be reabsorbed from the renal filtrate and sent back into the blood stream. the other conditions are transient.5 g/24hrs protein in urine. Due to the excess of glucose in the blood. If the blood glucose level exceeds 160 . galactose. Conditions or diseases associated with proteinuria: 1. trauma 3. Normal fasting blood glucose = 70 .120 mg/dl.05% sensitivity for sugars: Benedicts is overly sensitive. resulting in a coma and possibly death. The ketones in the blood are neurotoxic and will depress the activity of the brain. jaundice results. and fructose. may appear to be in a "stupor. fad diet 3. which tests for various reducing sugars in the urine. Symptoms of diabetes mellitus: polyuria: excessive urine output Polydipsia: excessive thirst Polyphagia: excessive eating There may be other conditions that result in hyperglycemia and glucosuria. Hypertension 2. orthostatic proteinuria. Average life span of RBC is 90 . sclera. including glucose. resulting in ketosis. and nail bed become yellow. and lapse into a coma. semen. and spasms 3.4. Diabetes mellitus: condition in which there is defective insulin or insulin is produced in reduced amounts. Moderate: 0. Non-specific. Where is insulin made? Insulin allows glucose to enter cells in the body.5 . pancreatic cancer Glycogen (liver) ---------------------> glucose Tests for "sugar" in the urine may be: 1. It detects acetone and diacetic acid.25% sensitivity for sugars 2. They are produced when carbohydrates cannot be used properly for energy." become unconscious. Occurs with: 1. not any reducing sugar. and infectious mononucleosis Urine Bilirubin and Urobilinogen Levels Health Prehepatic Posthepatic Hepatic urine bilirubin negative negative positive positive uriune urobilinogen normal positive decreased-normal positive . These tests are based upon the reduction of copper sulfate and are simple.0 mg/dl) But too much is neurotoxic. Ketones will build up in blood.5 g/24hrs protein in urine. they must go to another source.3. anxiety 4. It will not be affected by the presence of other reducing sugars. Proteinuria occurs due to glomerular damage: walls of capillaries become more permeable and allow large molecules to pass out in the urine. aspirin therapy 4.GLUCOSE IN THE URINE Glucosuria (glycosuria): the presence of glucose in the urine. diabetes mellitus 2. If insulin is lacking or defective. tumors. It is the most significant indicator of renal disease. Physical exercise 5.180 mg/dl. Except for diabetes mellitus. Small amounts of ketones in blood is normal (2.0 . associated with glomerulonephritis. Urinary tract infection (UTI) 5. exposure to cold Orthostatic Proteinuria: Protein present in urine after prolonged after prolonged periods of standing upright but absent after patient is laying down.10% glucose. associated with chronic UTI (of kidney). liver cancer. glomerulonephritis: glomerulus (capillaries) of kidney becomes inflamed and damaged and allow proteins to seep into the urine filtrate. dull. Even healthy individual may test trace + by Benedicts method. Ketosis in a diabetic occurs: 1." severe ketosis can develop. Starvation 4. About 1% of RBC's are destroyed each day and another 1% are made each day. False negatives occur: very dilute urine. failure to take insulin 2.

54 % Increase: low oxygen (packed RBC volume) Women 37 . Hodgkin’s Disease and bacteria infection 1 . short of breath. coagulation factors.5 x 106mm 3 Decrease: anemia using 1:200 dilution) 2. and plasma.3 % eosinophils Parasitic infection. Buffy Coat ( WBC’S and platelets) &RBC’s Layers of non anticoagulant blood (Red top). allergic reactions.0 . Coagulation factors.. add 10%.1 % basophils Allergic reactions and anemia/leukemia 8. viruses. sinusitis).90% of plasma is water 5. parasites. Water .000 . as well as their formation structure and function.0 .6.g. leukemia.0 g/dl Increase: low 0xygen binds to gases Women 12-16 g/dl Decrease: anemia makes up 95% of RBC mass made in bone marrow 4. Leukocytes ( WBC’S) nucleated cells with varying size and life span.5 um) macrocyte . asthma.45 7. septi(# of cells counted x 50 using 1:20 dilution . WBC’S.000/mm3 Increase: polycythemia and after splenectomy Decrease: anemia. or about 6 liters. and fibrinogen in one clot) See pictures below.000 .10% monocytes Monocytic leukemia. Acts as a buffer system to keep blood pH at 7. The total volume of blood makes up about 7 .36 % Increase: spherocytosis (MCHC) = Hb x 100 Decrease: iron deficiency anemia Hct 6.400. function to protect body from disease 2. Hemoglobin (Hb) Men 14. Platelet Count 140. fungi and other injurious agents to protect the body from disease.large RBC (>9 um) microcyte .proteins which specifically interact with bacteria. RBC Size normocyte RBC (7-7. IIMMUNOLOGY AND SEROLOGY NOTES Hematology is the study of blood components including the formed elements. Functions of Blood: 1. White Blood Cell Count 5.000/mm 3 Increase: infections (pneumonia. measles. Proteins .100 femtoliters (fl) Increase: newborns. Three layers of anticoagulant blood: Plasma (contains fibrinogen) . sickle cells polychromasia = increase in the amount of Hb in RBC’s which makes RBC’S appear blue instead of red. meningitis. and pregnancy (# of cells counted. Erythrocytes (RBC’S) non-nucleated cell with biconcave shape with 90 .35 .5. Hematocrit (Hct) Men 44 . fibrinogen are the major plasma proteins which function to maintain blood pressure and blood viscosity 8.8 % of an individuals body weight. Complete blood Count (CBC) The following test are done by an automated coulter counter machine in clinical labs). Red Blood Cell Count Men 4. *Anemia = a decreased number of RBC’s.000 Women 4. platelets. chemotherapy and radiation therapy 7. . RBC Indicies Normal Range Clinical Significance Mean Cellular Volume 80 . Red Blood Cell Morphology (see above). using 1:100 dilution.10.0 x 106 mm 3 Increase: congenital heart disease & low 0xygen (# of cells counted x 10. vit. Patient may be pale.47 % Decrease: anemia 5. multiply by 100 Decrease: some viral infections (mumps. and weakness. globulin.18. B12 & folate anemia (MCV) = Hct x 10 Decrease: iron deficiency *anemia RBC Mean Cellular Hb 27 . and skin disorders 0 . Test Normal Range Clinical Significance 1. dizzy. Thermoregulation 5. Nutrients/ Wastes 6. Serum (plasma minus the protein fibrinogen) and RBC Clot (RBC’S.120 day life span: function to carry 0 and other gases 2 3. Transports hormones from site of production to site of action Composition of Blood: 1.small RBC (<5 um) RBC Hb content normochromic (normal Hb) hypochromic decrease Hb hyperchromic increase Hb Terms associated with RBC morphology anisocytosis = abnormal variation in size of RBC’S poikilocytosis = abnormal variation in shape of RBC’S e. Antibodies.5 . trama.7. irregular heart rate. have headaches.albumin. Thrombocytes (Platelets) cytoplasmic fragments from parent cell megakaryoctye: function in blood clotting 4. White Blood Cell Differential: 100 cells are counted using Wright’s stain : Normal distribution: Distribution is increased due to: 50-65% neutrophils Bacterial infection and granulocytic leukemia 25-40% lymphocytes Viral infections and lymphocytic leukemia 4 . polio. appendicitis.counting 4 corner squares) cemia.Transport of oxygen from lungs to cells 2 Transport of wastes CO (HCO ) from cells to external environment 2 3 3.HEMATOLOGY. Transport nutrients to cells 4. counting all 9 squares) and AIDS) chemotherapy and radiation 3. Limits spread of injurious agents via inflammatory response 6.substance in blood enabling it to clot 7. and hematocrit.32 picograms (pg) Increase: spherocytosis (MCH) = Hb x 10 Decrease: iron deficiency anemia RBC Mean Cellular Hb Concentration 32 . hemoglobin.

and X) and coumarin therapy . People with allergies have increased IgE.fluorescent dyes are used to detect Ag-Ab reaction 6. heparin therapy and vitamin K deficiency Lee-White Clotting Time is manual coagulation test for PT and PTT. V.10 mm/hr Women 0 .a test to detect the yeast Cryptococcus Neoformans which causes meningitis.First Ab to rise during infection.5 . large RBC’S. These are treponemal test because true antibody produced in patients with syphilis reacts with coated antigens to form a reactive (positive) test C. VII. VDRL (Veneral Disease Research Laboratory) is another common non treponemal test done on CSF. monocytes. cirrhosis of the liver. Blood Bank and Serology antigen . and increased RBC’s (Erythrocyte Sedimentation Rate) Reticulocyte Count (young RBC’s with blue granules using stain) #reticulocyte counted X 100 = % reticulocyte # erythrocytes counted Prothrombin Time (PT) heparin 0. immunoglobulin (see below).a test to detect the heterophile Ab of patients that are infected with the Epstein-Barr virus. vitamin K deficiency. B . swollen lymph nodes.testing for rheumatoid factors that are in patients who have rheumatoid arthritis RPR (rapid plasma reagin) . also called. Also.uses patients serum with commercial cells. This is a non treponemal test because an antibody-like substance (reagin) that is produced in patients with syphilis reacts with a cardiolipin anitgen suspension to form agglutination (flocculation) . 1%) Increased: acute blood loss or response to treatment of anemia 11-13 sec Increased in clotting factor deficiencies (II.arc of precipitate is used to detect Ag-Ab reaction 4. small enough to cross placenta (see below). and O cells cross match -testing the donor RBC’s with the serum of the patient receiving the blood to make sure no reactions will occur Antibody Screen-a test performed to screen a patient’s serum for and Ab that might interfere with a RBC transfusion Transfusion Reaction . vaccines) or passively ( from mother at birth) direct or forward blood typing . ASO Screen .RBC’s are used to detect Ag-Ab reaction .antigen eating cell ( neutrophils.Reactive Proteins (CRP) . 1:64 etc) Hemolytic Disease of the Newborn . and other traumas. Hemaglutination Test . Immunoelectrophrosis Test . Precipitatin Reaction Test . so the reaction must have a way for visual detection of reaction. IgM (6%) . V. The symptoms include sore throat.a test to see how much antibody is in a patient which correlates to infection (ie.gel agar is used to detect Ag-Ab reaction 7. II. liver disease. complement . Also. heat at site of transfusion and / or death when ABO incompatible blood is given Antibody titer . IgE (0 -. asprin. or patients with tonsilitis. and eosinophils) natural immunity . too large to cross placenta. This can happen in childbirth complications. Crytococcal Antigen Test .mother has 2nd Rh+ newborn because the Rh+ antibodies was produced by the mother after the 1 st baby.a test to detect antistreptolysin 0 which is produced in patients with Group A Strep infections because the organism produces streptolysin 0. Latex Agglutination Test .electrical current is used to detect Ag-Ab reaction 3.anti-b or anti-d reverse or indirect blood typing . lower back pain . AB. (causes infectious mono). IX.Secondary immune response Ab (see below: shows previous infection or exposure).a test to detect inflammatory conditions Cold Aggultinins .light scattering is used to detect Ag-Ab reaction 5. emboli (floating blood clot). Rh Ab.a test used to screen for syphilis caused by Treponema pallidum.1.a test for systemic lupus erythematosus (SLE) and other autoimmune disorders.20 mm/hr Increased: inflammatory conditions (anemia. Treatment today is rogram. Activated Partial Thromboplastin Time 31-39 sec Increased in clotting factor deficiencies (I.a series of 11 serum proteins which helps fight off infection phagocyte . Nephelometry . and deficiency of a Duke method (puncture earlobe) 1-3 minutes coagulation factor (hemophilia) Capillary Coagulation 2-6 minutes DIC Panel (Disseminated Intravascular coagulation: Patient suffer form internal blood clotting throughout body.a test to detect primary atypical pneumonia caused by Mycoplasma pneumoniae. and exhaustion. tissue damage) Decreased: sickle cell.uses patients blood with anti-a . VIII.latex beads are used to detect Ag-Ab reaction 2. Infectious Mononucleosis . and powerful agglutinin and hemolysis (see below). MHA-TP or FTA-ABS . and staphylococcemia.Other hematology test not part of CBC Test Normal Range Clinical Significance ESR Men 0 . rash. chills. weakness. influenza or other acute respiratory infections. tumors. X. African trypanosomiasis. The antigen (Ag) antibody(Ab) reaction being detected in the aforementioned test are beyond microscopic examination.5% (ave.are tests used to confirm for syphilis caused by Treponema pallidum. XI. Bleeding Time Increased with throbocytopenia. IgD (0 .test for measles Rheumatois Factor Test . A. headache. pregnancy. 1:16.immunity acquired with out exposure acquired immunity . platelet Ivy method (puncture forearm) 1-7 minutes dysfunction. where the body attacks itself.Functions in allergic responses.A foreign substance that elicits an immune response antibody (Ab) -A protein substance developed in response to an antigen.When Rh.1%) Unknown function. actively (ie.A test to detect pregnancy by testing for the hCg hormones (Human chorionic gonadotropin) found in pregnant women and in abnormal conditions such as cancer of the ovaries and testes. Rubella HIA . Fluorescent Antibody Test . (APTT or PTT ) XII).an abnormal reaction by a patient that has recently been transfused which include fever. Rapid Immunoduffusion Test (RID) . IgG (80 %) . All serology test are done by one of the following methods: 1. ABO Ab.002%) . anxiety. Common serology test Antinuclear Antibody Test .antibodies in blood due to exposure. Pregnancy Test . gunshot wounds. 1:2. IgA (13%) found in saliva and GI tract to response to pathogens entering those areas.

From the liver. . renal disease. in the spleen. Normal adult range 2. Normal Range Concentrations in males 0. diabetes insipidus. Normal range: 8-18mg/dl(2.150 units/L Amylase 53 . diarrhea.9.Clinical Chemistry When analyzing a patient it is important to take steps to decrease random analytical variation (i.2 mg/dL (53 to 106) M mol /L) females 0. urea enters the blood and is ultimately excreted by the kidneys. Lactescent or Lypemic serum with increased triglyceride concentration will result in cloudy or turbid serum which will interfere with certain tests. diuretics Chloride 98-108mmol/L Increase: dehydration. vomiting. tobacco smoking. CK) 10-100 U/L Increased in heart attack Urea: Blood Urea Nitrogen (BUN) is the major end product of protein and amino acid catabolism and is generated in the liver through the urea cycle.increased in heart attack ( myocardial infraction) Creatine Phosphatase (CPK. and elsewhere.e. As bilirubin passes through the intestines it is converted into urobilinogen by bacterial enzymes.) Other factors that can change analytical test values are exercise. Important Clinical Chemistry Lab Tests Electrolytes: Testing Electrolytes are ions that exist in body fluids. or renal disease Potassium 3.4mmol/L) Decreased: starvation. loss of CO2 Decrease: acidosis of diabetes. Addison’s disease. and kidney disease. after steroids are used. Urobilin is a brown pigment formed by the oxidation of urobilinogen which may be formed in stool or urine after exposure to air.6 -1. The creatnine level is affected primarily by renal dysfunction.6 .is the break down product of purine ( which is the end products of nucleoprotein digestion). Increased: high protein diet. and water imbalance.123 units/L Cardiac (Heart) Enzymes. Normal range: Direct: up to 0. and thus very useful in evaluating renal function. CNS (maintains blood pH) depression. such as pneumonia. vomiting Bicarbonate 22-28mmol/L Changes in lung function. and dehydration. diet. and heart and muscle function. Creatine – is made in the liver from amino acids and used in muscle as an energy source. pregnancy. fever burns. Decrease(Hyponatremia): diarrhea.0 mg/dL (44 . anoxia and acidosis Decrease(Hypokalemia). pH. It is excreted entirely by the kidneys.2 mg/dL Uric Acid .88 mol/L) Creatinine – Creatinine is the waste product of creatine phosphate.0. a compound found in the sketel muscle tissue. shock.1.4mmol/L Increase(Hyperkalemia). physiologic effects of drugs and posture.4 mg/dL . Increased levels occur in renal disease. The major cation is Sodium (Na+) and Potassium (k+) and the major anions are Chloride (CL-) and bicarbonate (HCO3_) These four ions have great effects on hydration. prolonged fasting. Normal Range 0. diabetic ketosis and renal failure.most enzymes are increased in liver disease Test Normal Range Clinical Significance Alkaline Phoshatase (ALP) 20-105U/L Increased in liver disease Alaninetransaminse (ALT or GPT) 3-30 U/L ” Aspartate transaminase (AST) 6-25 U/L ” Lactate Dehydrogenase (LDH or LD) 125-290 U/L ” Pancreas EnzymesIncreased in pancreatitis Lipase 10 .9-6. The content of certain specimens may interfere with testing: Icteric serum with increase bilirubin will result gold yellow color and will interfere with certain tests.7 .Total: up to 1. Most uric acid is formed in the liver. low protein.5-5. ethanol. acid-base balance.5 -1. Test Normal Range Clinical Significance Sodium 135-148mmol/L Increase(Hypernatremia): dehydration. Liver Function Test Bilirubin – It is produced from hemoglobin of red blood cells by reticuloendothelial cells in bone marrow. by taking specimens at the same time of day with measuring test ran on daily basis.0 mg/dL Liver Enzymes.

0 mg/L Daily Requirement 800 mg Carbohydrate Testing Glucose (dextrose) .Blood and urine glucose testing every hour for up to five hours.Calcium . ect. Protein Testing . fat stores are metabolized which leads to the formation of ketones bodies. The body used glucose as a source of energy.180)mg/dL) glucose appears in the urine (glycosuria) which is a symptom of diabetes. Test Normal Range Clinical Significance Total Protein 6-8 g/dl Albumin 3. When the renal threshold (160 . D-xylose is used because it is not normally found in blood and urine. Serum globulins are antibodies. excitability of skeletal and cardiac muscle. Normal Range 9. tablets can be taken. Blood and urine levels are taken at 2 hours. neuromuscular conduction.is the most important carbohydrate in body metabolism. preservation of cell membrane integrity and permeability. Calcium plays a vital role in such basic physiologic processes such as blood coagulation. Norma blood serum 80 . Low Calcium or loss of Calcium salts due to vitamin D deficiency can result in osteoporosis and osteomalacia (softening of bones). but increased in diabetic patients.Testing glucose levels two hours after a patient’s meal.increased in blood glucose levels. D-xylose Tolerance Test .Two major protein groups are Albumin (about 60% of total protein) and globulins (40% of total serum proteins). Phosphorus also aids in structure of bones and teeth. Albumin help to maintain fluid balance. Levels are rarely elevated in normal patient. and increased proteins.5 umol/ L) Increased in lead poisoning.A test to detect if patient is deficient in a mucosal lactase enzyme that breaks down lactose into the simple sugars. Iron Testing 65-165 mg/dL (11. These patients experience gastrointestinal discomfort which may be followed by diarrhea.(C++) the most abundant mineral element in the human body and approximately 98 % of calcium in the adult is present in the skeleton.11. Failureof the pancreas to produce adequate insulin (which allows glucose to enter the cells of the body. but can be found in fruits such as plums. Insulin testing . also a symptom of diabetes. and needed for lactation in females. results in hyperglycemia . A normal patient should have normal glucose level after 2 hours. acid from intestines. growth hormones. kidneys or GI tract.120 mg/dL. maintenance of normal tone.6-29.2 . Lab Test Cholesterol Testing 140-250 mg /dL (< 200 is best) Increased in coronary artery disease. then giving the patient a standard load of glucose. Decreased in iron deficiency anemia due low intake of iron Hormone Testing Thyroid testing. then monitored for 5 hours.5 -5. after taking a fasting glucose level. The remaining calcium is present in extra cellular fluid and skeletal muscle. Misc. nutrition and liver function since most serum proteins are made in liver. Milk should be removed from diet or lactiad. Parathyroid testing Male & Female hormones.2 g/dl Decrease in albumin may occur in liver disease starvation and protein loss through skin. The patient in given D-xylose. Lactose Tolerance Test . Glucose Tolerance Test . Postprandial Glucose Test . Calcium absorption is enhanced by vitamin D. blood coagulation proteins and enzymes can detect state of dehydration. When the blood sugar is below normal (hypoglycemia). glucose and galactose.A test to see if absorption is taking place in small intestine because sometimes bacterial overgrows in the small intestine affecting absorption in the jejunum.

just inside the cell wall . grow best at T between 20 C.MICROBIOLOGY NOTES The Bacterial Cell Bacteria are named by genus and species.reduces growth Control of Microbes with Heat 1.g. Staph. Example: Colistin Nalidixic Acid (CNA) .5%) allows the medium to be selective for Staphylococcus since most organisms cannot tolerate that high salt content. flagella. but most at 7.closed circle of double stranded DNA which contains a. and site of action for many e. Moist Heat. Staph.1. peritrichous . Temperature requirements ( refrigeration.0.(autoclave) kills vegetative forms and spores achieves 100% kill. aerobic organism (aerobe) = require O2 for growth b.flagella over the entire surface of bacteria 5.Enhance the growth of most gram positive organisms. A selective. microaerophilic organism = require small amounts of O2 d.determines gram reaction (purple positive.normal human body temperature 300. Salmonella Shigella Agar (SS) 1. mesophiles= like mild temperatures.site of protein synthesis bacteria.thin. atrichous . thermophiles= hot lovers. amphitrichous .Media that allows the growth of groups of organisms and demonstrates certain characteristics. 2. epidermidis appears pink MacConkey Agar (MAC)) . Oxygen Requirements: a. 1. MRVP. Gram positive organisms and coliform organisms are inhibited by the bile salts mixture. made mostly of water. Gram positive organisms and coliform organisms are inhibited by the bile salts mixture.5% agar ( the gelling agent) . Lactose fermenters are hot pink to red. Genus begins with a capital letter.liquid matrix of the cell. Endospores. Antiseptics . anaerobic organism (anaerobe) = can't survive in presence of O2 c. grows many organisms 3.group of flagella at one pole 4. psychrophiles= cold lovers. Colistin and nalidixic acid is the inhibitor agent that inhibits the growth of gram negative organisms. Sterilization . Streptococcus pyogenes a. Common enrichment media 2. 100oC) 3. 4oC slows bacterial growth.g. non-lactose fermenters form colorless or very light pink colonies. e. 4.green zone due to partial destruction of RBC surrounding colony Beta ( ). Simmons Citriate. 2. Nutrient Broth (N Broth) . petri plates or test tubes semi. non-lactose fermenters green colonies. Growth of most microorganisms in inhibited by solutions whose osmotic pressure is more than 1%. differential medium for the isolation of gram negative rods (bacilli). 3. They are a smaller than the chromosome. some bacteria. acidophile= grow best at 5. neutrophiles= grow best at near neutral pH of 6.contains 1. Hektoen Enteric Agar (HE) 1. It may be transferred from one cell to another environmental conditions are unfavorable.regulates exchange of materials ribosome’s (40s + 60s) into and out of the cell 9.clear colonies with black center. Gram positive organisms and coliform organisms are inhibited by the bile salts mixture. some don't. pili. a. species begins with a lower case letter and both are underlined.7% agar.Media that contains chemicals or dyes that prevent or retard growth of certain organisms and select or allow growth of other organisms without necessarily differentiating them. 3. 2. shows hemolysis Hemolytic Patterns: Alpha (  ).8. Bactericidal 100% killing of microbes 5. Sabourad Dextrose Agar.1. polar or monotrichous .most organisms will not grow 42o except for Pseudomonas aeruginosa Factors Affecting Microbial Growth 1. tuft or lophotrichous . 4.0 or less. TSA (nutrient agar) with 5% Sheep Blood (Blood Agar Plate. aureus appears yellow. non-lactose fermenters form colorless or very light pink colonies. Lactose fermenters are yellow 4. Lactose fermenters are hot pink to red.an enrichment medium for the isolation of fungi (yeast and molds) Differential Media. for attachment to other cells (human/bacteria) or to other surfaces. The high salt concentration (7. Osmotic Pressure Requirements Practical applications: ( use of salts to pressure foods. MAC Agar does not. test tubes Media Definitions and Examples Enrichment Media. pink negative) b.clear zone due to total destruction of RBC surrounding colony Gamma ( ). 3. not just the surface. nuclei. plasmid. Steam under pressure.Media used to identify microorganisms. Examples: Salmonella. Dry Heat-( cooking food) must cook food throughout to kill microbes.reduces microbial population. and TSI Temperature and technique employed in the incubation of primary culture Most organisms multiply best at temperatures similar to those in the host: 350. grow best at -50C through 200C b. 3. Humans have 80s of lipids .no Hemolysis Chocolate Agar (Choc) . HE Agar shows H2S production.a small piece of extra chromosomal DNA.gives bacteria its shape. It is used for the Kirby. permeable membrane 8. alkylophiles= grow best in an alkaline environment of 9. Agar Mueller Hinton Agar (MH) .temperature of the surface of the body.Basic media containing additives such as blood or serum which enhance the growth of many fastidious and non-fastidious organisms. ( most fungi are incubated at 300) 42o. 15 minutes .Clear colonies without black non-lactose fermenter. a. MAC Agar does not. freezing slows growth even more) All bacteria have a To at which they grow best and have their shortest generation time = T optimum. lribosome.single or tuft at one or both poles nutrients and wastes e. pH requirements Practical Application of pH and growth of bacteria: Use of weak acids to preserve foods All bacteria have a pH optimum a pH at which they grow best and have the shortest generation time. use of sugars to preserve foods) Osmotic Pressure is the concentration of dissolved sugars or salts in a solution.proteins embedded in it . yeast and molds b.reduce microbial growth. cytoplasm.solid-contains 0.0 or more 3. Example: Mannitol Salt Agar (MSA) . dissolved d. Selective Media. Shigella.one flagella at one pole the genetic information of the cell b. i.Bacteriostatic. like ToC 45oC and above o o c. cell wall . Biochemical Media .Disinfecting .(boiling. but not a 100% kill.BAP). chromosome.1. 15 lbs psi.e.An enrichment medium of many organisms especially Heamophilus sp. solid.: intestinal bacteria antibiotics . A selective. sterilization at 121oC.Bauer sensitivity test. Examples SIM.An enrichment medium for many kinds of organisms. It is much 10. and is double resistant form of the microorganism and form when the stranded.0. SS Agar shows H2S production.100% killing of microbes in or on a surface 2.responsible for bacterial motility 3. differential medium for the isolation of Salmonella and Shigella which is a gram negative rods from feces or rectal swabs. but not a 100% kill. Exception: halophiles: require high salt concentration to grow and reproduce Control of Microbes 1.hair-like structure extending from the surface of the cell and Clostridium tetani Media Media is any substance that contains all the necessary components for growth and reproduction of bacteria Basic Media Types Liquid (broth) test tube .without flagella 6.: Clostridium botulinum 7. A selective. Most differential media is also selective.0 c.some bacteria have them. used to isolate staphylococcus species. differential medium for the isolation of Salmonella and Shigella which is a gram negative rods from feces or rectal swabs. non-lactose fermenter (H2S). and Neisseria gonorrhea which will only grow on Choc. 4.area in the cytoplasm where the chromosome is located c. to exchange plasmids: sex pili used in a process called conjugation 2. cell membrane . 3. facultative = can grow in the presence or absence of O2 2.45oC 4. 2.Grows many organisms including anaerobes and can recover small amounts of organisms in cultures.

. b. X & V Differential -To identify Haemophilus species. Acquisition of infectious microorganisms 1.5 Macfarland concentration of bacteria. Inoculate plates. Perform antibiotic sensitivity test (see picture 25) to determine if organisms are sensitive or resistant to various antibiotics: (A) MH Agar plates are completely covered with a . swarming colonies on blood or choc. TSI -To identify various gram negative rods. Crystal Violet. and Enterobacter) from Pseudomonas species.decolorize (remove purple stain from certain organisms) ****DO NOT LEAVE ON TOO LONG OR ALL ORGANISMS WILL LOSE PURPLE STAIN AND END UP STAINING RED**** e. Gram stain examinations should be done to determine: a. 3. Contact with infected or colonized material: (human to human. agar c. Aerosols. Iodine. Heat fix slide to make bacteria adhere to the slide. gross colony morphology. coli. and streak plate to get good isolation by flaming loop between each quadrant 2. scarlet fever. vectors (diseases transmitted by insects. nose. vaginal infections.) arthropods. .stains red/pink. (group A strep) and septicemia (bacteria in the blood which causes high fever) Streptococcus pneumoniae pneumonia Staphalococcus aureus boils. 5. in fact they are beneficial to the host by stimulating development of the immune system and interfering with the establishment of population of pathogenic bacteria. The carrier state: Individual with no signs or symptoms of disease may be colonized by pathogenic microorganisms and may transmit the pathogens to susceptible hosts who develop clinical disease.positive (purple) vs. 7. Decolorizer. 2. 4. Resistant = organisms will grow up to the antibiotic disc because the antibiotics did not inhibit the organisms from growing around the disc which means that the doctor could not use this drug to treat the patient infected with that bacteria.Common biochemical test: Catalase -to distinguish staphylococcus species from streptococcus species. Opportunistic infectious processes often are caused by microorganisms that are considered to be nonpathogenic for health individuals. flat colonies on MAC Agar b. ingestion of contaminated food and /or drink. Bactitracin (“A” disc) -to detect Group AB Streptococci. carbuncles.e. (B) Antibiotic discs are dropped on agar plates and incubated for 24 hours. and septicemia Escherichia coli urinary infections rotius mirabilis urinary tract infection Pseudomonas aeruginosa respiratory & urogenital infections Salmonella enteritis typhoid fever Neisseria gonorrhea gonorrhea Clostridium botulinum botulism (food poisoning) Klebsiella pneumoniae pneumonia Fungi ring worm . SIM. 5. Bile Esculin only -to detect Group D Streptococcus (not Enterococcus). Bile Esculin and Salt -to detect Group D Enterococcus. fomites (inanimate objects contaminated with microbes). morphology (shape of bacterial cell) Principle of GRAM STAIN STAINS USED TO MAKE THE BACTERIA CELLS VISIBLE a. Klebsiella. color of colony d. and by providing beneficial vitamins Colonization. urinary tract and vagina). Citrate. Oxidase -to distinguish Enterobacteracae species (E. Coagulase -to distinguish staphylococcus aureus from staphs epidermidis and other staphylococcus species. gram stain reaction .shortly after birth.stains organisms that did not retain the purple stain red/pink GRAM POSITIVE ORGANISMS-. They did not retain purple dye but took up the red counterstain. Culture quantity should be done 5. etc. and look closely for mixed organisms.stains organisms purple c. or systemic. Negative (pink) b. PATHOGENS DISEASE Streptococci pyogens strep throat. mouth. Safranin. MR-VP.mucoid vs. and athlete’s foot Haemophilis influenza meningitis ear infections Enterococcus faecalis wound infections and urinary tract infection (group D strep) Corynebacteium diptheriae causes diptheria. animal to human. Nosocomial infections are acquired in a hospital or health-care setting . The immunosuppression leading to opportunistic infection may be transient. elevation .Interpretation of Cultures 1. Hold Blood Agar Plate up to the light to look for hemolysis.enhances purple stain d. usually as the result of debilitation and temporary immune incompetence. Optochin (“P” disc) -to detect Streptococcus pneumoniae. form and elevations of colony 6. gastrointestinal tract. Colonies should be examined for characteristics such as: a. rheumatic fever. Lactose Fermentation -Determine which biochemical test to be done or setup on gram negative rods. They retain purple dye.bacteria that is on almost all surfaces of the human body that do not harm by their presence. infection forming a false membrane on any mucus surface Normal Flora Normal Flora . Set up biochemical test . GRAM NEGATIVE ORGANISMS-. (see picture in book). After incubation observe colonies for growth. Opportunistic infections. most surface areas of the human body are colonized with normal flora (i. localized. (C ) Plates are read as follows: Sensitive = organisms will not grow up to the antibiotic disc because the antibiotics inhibited the organisms from growing around the disc which means that the doctor could use this drug to treat the patient infected with that bacteria. and flies 3. 4.pneumonia. margins. . pharynx. skin.individuals who are immune suppressed by other natural or chemical means are more susceptible to infections.stains purple.

Olds. 6.References for Notes and Labs 1. Norma.. Anna. Publishers .. Polko. New York: McGraw. Paul: West Publishing Co. Raul. Medical Laboratory Procedures. R. Medical Laboratory Procedures. 2. 1999. Basic Techniques in Clinical Laboratory Science. Medical Laboratory Assistant. 4. Color of Microbiology.Instructors Manual.Hill Publishing Co. 2nded. Basic Microbiology. Inc.. Reynolds. Jacquelyn. St. Barbara. Wesley. Volk. Inc. Hilda. Chicago: Year book Medical Publishers. New York: McGraw. Hilda. Tom. Walters.. Marshall. New Jersey: PrenticeHall.Inc. Jean. Cano. 1999. Estridge. New York: Harper and Row.1990. Microbiology. 3rd ed. Tom. 8. Palko. Basic Medical Laboratory Techniques.1990. Polko. Palko. 6th ed. New York : Delmar Publishers. St Louis: Mosby Yearbook 3.Hill Publishing Co. 5. Linne. 7.