Aubf Trans Prelim

OUR LADY OF FATIMA UNIVERSITY
MEDICAL LABORATORY SCIENCE BATCH 2024

ANALYSIS OF URINE AND BODY FLUIDS
MR. ROJOHN SONNY C. CRUZ, RMT
ADAPTED FROM: POWERPOINT/LECTURE
TRANSCRIBED BY: MA. LORAH YZABELA L. TUASON
COURSE OUTLINE: PRELIMS QUALITY CONTROL

1. Introduction to Urine  Sum of efforts to achieve the highest degree of
2. Analysis of Urine and Body Fluids excellence in order for the patient and physician to
3. Renal Physiology obtain accurate and precise results in the shortest
4. Chemical Examination of Urine possible time in a reasonable cost
 Factors involved in QC in C
REFERENCE BOOK
 Standard
 Controls
 Continued education
 Attitude - Motivation
 Equipment
CLINICAL MICROSCOPY
 Reagents
One of the oldest laboratory method that is an axillary
 Laboratory personnel
branch of laboratory medicine which deals with the study
of physical, chemical and microscopic study of body
SPECIMEN COLLECTION
fluids.
Considerations in specimen collection and handling
 Adequate volume of the sample
INTRODUCTION TO URINE  Medication taken should be noted
HISTORY  Avoid collecting during menstruation
 Hippocrates - “uroscopy” 5th Century  Observe universal precaution
 1140AD – color charts was developed (20 color)

where chemical testing progressed from and the QUALITY CONTROL
“TASTE TESTING “ for glucose and “Pisse Some laboratory rules
Prophets”  All laboratory request must be filled up properly
 Frederik Dekkers – albuminuria by boiling 1694 and correctly
 17th Century – examination of urine sediments  Proper Specimen collection, transport and
 Thomas Addison – method for quantitating urinary storage
sediments  Maintenance of record books and accession
 Richard Bright – 1827 introduced urinalysis as part of books
doctor’s routine  Maintenance and calibration of equipment and
 Popularity of urine laboratory materials
 Availability  Material safety data sheet
 Source of information on body’s major metabolic  Laboratory Safety
functions
SPECIMEN COLLECTION
SIGNIFICANCE  Specimen must be collected in a clean dry container
Various diseases/ disorder can be monitored by  Specimen must be properly labeled –
urinalysis due its composition such as  Specimen must be delivered to the laboratory
 Body metabolism diseases promptly and must be processed within 2 hours
 Endocrine function  Preservation
 Renal function  Physical
 Chemical
 Purpose of preservatives • Formalin 40% - 1 drop/ 30 mL of urine; is an excellent

 Suppress bacterial growth preservative for sediments; interfere with Obermayer
 Which converts urea to a urea splitting test for indicans, Fehling’s and Clinitest, may precipitate
enzyme with urea thus interfere with microscopic examination,
 That degrades glucose may cause false ring w/ Heller’s test for albumin
 That interfere with protein test
• Formaldehyde tablet – 1 tab/ 60 mL urine; slightly
 Prevents instability of urinary solutes
increase specific gravity of urine
 Prevents degradation of orgaized sediments such
as PUS, cast etc 3. Chemical Preservatives: special
 If preservatives is added should be indicated on the • Conc. Hydrochloric acid – for epinephrine,
results noradrenaline, catecholamines, vanilymandelic acid,
steroids, ammonia, urea, total nitrogen
CHANGES IN UNPRESERVED URINE • Glacial Acetic acid – (pH 4.5)for aldosterone; (pH 2) for
 Increase bacterial serotonin
• Sodium Carbonate – for porphyrine and urobilinogen
 Increased pH
• Chloroform – aldosterone
 Decrease glucose • Sulfuric acid – preserves calcium and inorganic
 Decrease ketones constituents
• Sodium fluoride and benzoic acid – glucose analysis
 Decrease bilirubin
• Phenol – causes change in odor;
 Decrease urobilinogen
 Increase nitrite URINE COLLECTION TECHNIQUES
 Increase turbidity
Bottle method – a method that uses any receptacles to
 Disintegration of cast and RBC’s
collect the specimen provided that it is dry, clean, and
 Change of color sterile
Gauze-pad method- a gauze pad is used to collect the

METHOD OF PRESERVATION
urine and then centrifuges in a centrifuge tube with a golf
tee that will hold the gauze at a distance from the bottom
1. Physical
of the tube thus allowing clean urine to collect at the
• Refrigeration prevents the growth of bacterial and
bottom of the tube
helps preserve cast, RBC Pus, Epithelial cells. It
maintains an acidic pH for short period of time (8hrs)
Catheterization method-a rubber tube is inserted through
the urethral orifice to the urethral canal into the bladder
2. Chemical Preservatives: common
to collect a presumably pure urine specimen.
• Toluene – all around preservatives; 2 ml/ 100 mL urine;
forms a film that floats on the surface of the urine that
Supra pubic aspiration method – direct puncturing of the
keeps the contaminants out but not effective on bacteria
supra pubic region to collect urine directly from the
and molds already in the urine
urinary bladder.
• Thymol crystals – preserves glucose and sediments

Mid stream catch method – collection of urine specimen
well; but may give false + on albumin test and may
for examination at the middle part of a single continued
interfere with bile test
normal urination.
KINDS OF URINE SAMPLE/ SPECIMEN

ANALYSIS OF URINE AND BODY
1. SINGLE URINE SPECIMEN
FLUIDS
 Random Specimen – used for routine screening; any
urine sample collected any time of the day.
 Advantage: allows detection of pathologic post RENAL PHYSIOLOGY
prandial concentration of solutes
 Disadvantage: variation in dilution and FUNCTIONS OF KIDNEYS
concentration
1. Excrete waste products of metabolism
 First Morning Specimen - voided upon waking up;
2. Regulate acid-base Balance
ideal sample because of concentration w/ higher
3. Regulate Electrolyte balance
osmolarity and more acidity
4. Regulate Blood pressure
 Fasting Specimen / Second morning Urine Specimen
5. Regulate red cell production (erythropoiesis)
– second voided urine specimen after a period of
fasting. This specimen does not contain any
metabolites from ingestion prior to beginning of the
fasting period
 Suprapubic specimen - provides sample that is
sterile good for culture and cytologic examination
 Three Glass Collection – used to determine prostatic
infection; uses three containers then collecting the
first part of a single continuous urination, then
collectng the middle part on a 2 nd container, then
the last part with a 3rd container.
 Pediatric Specimen – Afternoon urine - A specimen NEPHRON
collected between 2-4 pm used for urobilinogen • Functional unit of the kidney
determination • 1-1.5 millions of nephrons in each kidneys
2. TIMED URINE SPECIMEN A. Glomerulus

 24 hours – used to measure total amount of solutes B. Renal tubules
present in the urine • Proximal Convoluted Tubules
 12 hours – used for a\Addis count • Loop of Henle
 Post prandial – collected 2 hours after meal; used to • Distal Convoluted Tubules
determine the presence of glucose and protein
 GGT specimen - fasting; ½ hour; 1 hour; 2hour; and
3 hour specimen; test for glucose and ketones to
identify renal threshold for glucose.
URINE FORMATION • Passive Transport- movement of molecules across a
1. Renal Blood Flow membrane as a result of differences in their

concentration or electrical potential on opposite sides
 1200 ml/min (depends on body size)
of the membrane.
 Renal Plasma flow 600-700 ml/min
Ex. Water,Urea,Sodium
2. Glomerular Filtration
4. Tubular Secretion
Factors that influence filtration process
• Involves the passage of substances from the blood in
A. cellular structure of the capillary walls and Bowman’s
the peritubular capillaries to the tubular filtrate
capsule
3 LAYERS
2 major functions
- capillary wall membrane
1. Elimination of waste products not filtered by the
- basement membrane
glomerulus
- visceral epithelium (podocytes)
2. Regulation of acid-base balance
B. hydrostatic pressure
C. oncotic pressure
SPECIMEN COLLECTION AND HANDLING
D. renin-angiotensin-aldosterone system
- controls the regulation of the flow of blood to and • Clean dry container
within the kidneys • Label patient’s name, date and time of collection
- responds to changes in blood pressure and plasma • Examine within 1 hour (not more than 2hours)
content • Preservation - refrigeration, freezing, chemical
 Renin production = low plasma pressure and DRUG SPECIMEN COLLECTION

plasma sodium • Chain of Custody
 Angiotension II corrects renal blood flow by: • Required amount-30-45ml (DOH 60 ml )
 Vasoconstriction of renal arterioles • Urine Temperature -32.5-37.7 C within 4mins from the
 Proximal Convuluted Tubules reabsorption of time of collection
sodium
 Release of aldosterone (Na retaining SPECIMEN EVALUATION
hormone) from the adrenal cortex • For single specimen submitted for multiple
 Triggers release of Anti Diuretic Hormone measurements, bacteriologic exam should be done
 Glomerular Filtrate-specific gravity 1.010 pH first 50 ml disposable container = 10-15 ml urine
7.4
 GFR= 120 ml/min PHYSICAL EXAMINATION
A. Color – varies,metabolic function,physical activity, diet
3. Tubular Reabsorption
 Normal = yellow
Cellular Transport mechanisms
 Yellow Pigment - urochrome named by Thudichum
• Active transport - substances to be reabsorbed
in 1864
combine with a carrier protein in the membranes of
 Orange Brown - Urobilin
the tubular cells.
 Pink to Red Pigment – Uroerythrin
• Electrochemical energy - produced by this interaction
transfers the substance across the cell membrane
Ex. Glucose, amino acids, salts, chloride, Sodium
Polyuria
URINE COLOR CHANGES WITH COMMONLY USED
• Increase Urine Volume
DRUGS
• Volume is more than 2,000ml/24hr
Levodopa(Ldopa) Red then brown • Diabetes Mellitus
• Diabetes Insipidus
• Increased salt intake and high protein diet
Metronidazole(Flagyl) Reddish Brown
• Drugs-caffeine, alcohol, thiazides, and other diuretics
Nitrofurantion(Furadantin) Brown Yellow • Intravenous saline or glucose solution

• Chronic Progressive renal failure
Phenozopyridine(Pyridium) Orange-Red(Acid pH)
Nocturia
Phenindione anticoagulant Orange/Red
• More than 500ml with a specific gravity of less
Rifampin Bright Orange Red than1.018 at night
Riboflavin Bright Yellow Oliguria

• Excretion of less than 500ml of urine daily
B. Transparency/Clarity
Anuria
 Bacterial Growth - uniform opalescence that is not
• Complete or total suppression of urine formation
removed by filtration nor acidification
 Leukocytes - white cloud(remains after acidification)
Residual Urine
 Pink Cloud - Urates
• Urine that is left in the bladder after voluntary urination
 Orange Cloud - Uric Acid
SPECIFIC GRAVITY
C. Odor
• Used in assessing the kidney’s ability to reabsorb
 Normal Odor- faint aromatic
• Detects possible dehydration or abnormalities in ADH
 Urinoid- Substance responsible for urine odor
secretion
• Aids in evaluating the concentrating and diluting
D. Volume
abilities of the kidneys
Average
• Clinical Corrections:
• Adults= 1,200-1,500 ml
• Normal value average 1.003
• Range = 600- 2,000
• Range 1.035
• Night Urine in General does not excess
• Isosthenuric- below 1.010
with a volume of 400ml
• Hyposthenuric- above 1.010
• Hypersthenuric- above 1.010
Factors that influence urine volume
• Excretion of radiographic contrast media and dextran
1. Fluid Intake
will give a very high urine specific gravity reading
2. Fluid Loss
(over 1.035)
3. Variations in secretions of ADH
METHODS: Harmonic Oscillation Densitometry
1. Urinometry - uses urinometer or hydrometer • Sound wave frequency
• Principle : water displacement or buoyancy • Automated instruments
• Disadvantage: requires 10-15ml

• Accuracy maybe checked by measuring the specific Falling Drop
gravity of • Timing the fall of a drop of body fluid of known size,
• Distilled water: 1.001 through a definite distance in a mixture
• Potassium Sulfate: 1.015 • Heavier drop will fall faster
• Specimen cold- subtract

• Specimen warm- add Osmolality
The concentration of a solution in terms of osmoles of
Reading is affected by: solute per kilogram of solvent

Osmometers:
A. Temperature – subtract 0.001 from the reading for
- Freezing point of osmometers
every 3 degrees centigrade that the specimen
- Vapor pressure osmometers
temperature is below the urinometer temperature
- add 0.001 to the reading for every3 degrees C that the
specimen temperature is above the urinometer
temperature
B. Glucose-subtract 0.004 for every gram of glucose/dL
C. Protein- subtract 0.003 for every gram of protein/dL
Dilution of urine - multiply the decimal factor by the
dilution factor to get the actual specific gravity reading.
2. Refractometry
• Refractometry determines the concentration of
dissolved particles in a specimen by measuring
refractive index
• Refractive Index- comparison of the velocity of light in
a solution and velocity of light in air
• Total Solids Meter
Refractometer maybe calibrated using the ff:

A. Distilled water: 1.000
B. 5% Nacl: 1.022
C. 9%sucrose: 1.034
Advantages:
1. Uses small amount of urine(1-2 drops)
2. Simple to operate
3. Gives Rapid Reliable Results
CHEMICAL EXAMINATION OF PARAMETERS OF DIPSTICK TESTING:

o Glucose (30secs) – Double sequential
URINE enzymatic reaction
- The most conventional method of urine o Bilirubin (30 secs) – Diazo reaction
chemical analysis is carried out through the use o Ketones (40 secs) – Na Nitroprusside Test
of a chemical impregnated plastic strip called (Legal’s Test)
REAGENT STRIP. o Specific Gravity (45 secs) – pKa change of
- Simple and rapid polyelectrolytes
o Proteins (1min) – Protein (Soresen’s)
REAGENT STRIP TECHNIQUE error of indicator
1. Dip the reagent strip briefly into a well-mixed o pH (1min) – Double indicator system
uncentrifuged urine specimen. o Blood (1min) – pseudoperoxidase activity
2. Remove excess urine by touching the edge of of hemoglobin
the strip to the container as the strip is o Urobilinogen (1min) – Ehrlich’s reaction
withdrawn o Nitrites (1min) – Greiss’s reaction
3. Blot the edge of the strip on a disposable o Leukocytes (2mins) – Leukocyte estarse
absorbent pad.
4. Wait for the specified time for the reaction I. SPECIFIC GRAVITY – assessment of kidney’s
to occur. concentrating ability.
5. Compare the color reaction of the strip pad - Defined as the density of a solution compared
to the manufacturer’s color chart under a w/ the density of a similar volume of distilled
good lighting condition. water at a similar temperature.
- Infulenced by the number and density of
CARE OF THE REAGENT STRIPS particles dissolved in a solution.
a. Store with desiccant in an opaque, tightly
closed container. CAN BE MEASURED USING:
b. Store below 30C (Cool dry place); do not 1. Urinometer (Hydrometer)
freeze. 2. Refractometer
c. Do not expose to volatile substances 3. Harmonic Oscillation Densitometry
d. DO not use past expiration dates 4. Chemical Reagent Strip
e. Do not use discolored pads
f. Remove strips only immediately before use A. URINOMETER (HYDROMETER)
and tightly reseal. - Consists of weighted float attached to a
scale that has been calibrated in terms
QUALITY CONROL OF REAGENT STRIPS of urine sp/gr.
 Always check with a (+) and (-) controls a - Calibration is done with K2SO4 (20.29g)
minimum of once every 24 hours. (or every to 1L of H2O (sp/gr = 1.015)
shift) - DISADVANTAGES: Requires large urine
 QC must also be performed every opening of volume, affected by TEMPERATURE,
a new bottle; when there is a questionable GLUCOSE and PROTEIN.
result; when there is a concern in the integrity RESULT CORRECTIONS:
of the strips. *Calibrated at 20C
 Dipstick reagent box insert must always be For every 3C rise, add 0.001 to the result
consulted for QC and QA. For every 3C drop, substract 0.001.
*For every 1g/dL of PROTEIN substract 0.003 to
2 MAJOR DISTRIBUTORS: sp/gr.
1. Rouche - Chemstrips *For every 1g/dL of GLUCOSE, substract 0.004 to
2. Siemens - Multistix sp/gr.
Sample problem: II. pH – important indicator for identification of

The urine sp/gr is determined using urinometer at crystals and determination of unsatisfactory
26C and read 1.025. What is the corrected value? spx.
NORMALLY:
B. REFRACTOMETER – principle is Refractive o Random Catch: 4 to 8 pH
Index or RI. o First Morning: 5 to 6 pH
- R.I. = comparison of velocity of light in air o Afternoon: > 7pH
w/the velocity of light in a solution. REJECT SPECIMEN IN pH IS 9.
- Compensated to temperature (no need for
correction) Principle: double indicator system
- But still requires correction for Glucose and Bromthymol Blue (reacts if pH is Basic)
Proteins. Methyl Red (reacts if pH is Acidic)
*Glu and CHON are High molecular substances
and does not relate to renal concentration ability CAUSES OF ACIDIC URINE:
but will increase specific gravity. - High Protein Diet -
CALIBRATORS: - Dehydration
o 5 % NaCl = 1.022 +/- 0.001 - Cranberries
o Distilled Water = 1.000 - Diarrhea
o 9% Sucrose = 1.034 +/- 0.001 - Diabetis Mellitus
- Acid (+) bacte
Sample problem: - Starvation (ketone build up)
Refractometer reading of sp/gr is 1.025. There are - Drugs
2g/dL of Glu and 3g/dL of CHON. What is the Methamine mandelate
corrected value? Fosfomycin tromethamine
C. HARMONIC OSCILLATION DENSITOMETRY CAUSES OF BASIC URINE:

- Based on the principle that the frequency of a - Renal Tubular Acidosis
soundwave entering a solution changes in - Hyperventilation
proportion to the density of the solution. - High Fiber (Veggies) Diet
- EX. YELLOW IRIS (international Remote Imaging - Urease (+) bacte
System) – 300 to 500 workstations - After meal
6mL of Urine = 4mL for IRIS slideless - Old specimen
microscope
2mL for IRIS Mass Gravity Meter III. PROTEINS – most indicative of renal disease
- White foam (albumin) upon shaking
D. CHEMICAL REAGENT STRIP - Normally <10mg/dL or 100mg/24hrs.
Principle: pKa change of polyelectrolytes (albumin)
Reagents: Principle: Protein (Soresen’s) Error of Indicator
MULTISTIX: Poly (methyl vinyl ether/maleic Reagents:
anhydride) bromthymol blue MULTISTIX: Tetrabromphenol blue
CHEMSTRIP: CHEMSTRIP: Tetrachlorophenol
Ethyleneglycodiaminoethylethertetraacetic acid tetrabromsulfonphthalein
bromthymol blue *Yellow indicator if (-)
- Test is sensitive to the number of ions in the *Blue to Green color if (+) SENSITIVE TO ALBUMIN
urine specimen; indicator changes in color in
relation to ionic concentration. PROTEIN CONSTITUENTS OF URINE
Blue  Green  Yellow 1. Albumin – major urinary protein
2. Serum Tubular Microglobulins
*affected by pH – add 0.005 to the sp/gr if
3. Tamm-Horsfall Protein (Uromodulin)
pH is >6.5.
4. Proteins for prostatic discharge/vaginal **ORTHOSTATIC PROTEINURIA – due to increased pressure

secretion. on the renal veins
Orthostatic Clinical
CLINICAL SIGNIFICANCE Proteinuria Proteinuria
1. Protein detection require additional st
1 morning urine - +
testing Random Urine + +
2. Clinical proteinuria > 30mg/dL
b. TUBULAR PROTEINURIA
CATEGORIES: Pre-Renal, Renal and Post-Renal - Indicates disorders affecting the renal
A. PRE-RENAL PROTEINURIA – caused by tubules.
conditions affecting the plasma prior to its - Impaired reabsorption.
reaching the kidney. CAUSES:
- Not indicative of actual renal damage - FANCONI SYNDROME
- Not detected by reagent strip for CHON - Toxic agents
because it only detects ALBUMIN. - Severe viral infections
CAUSES: C. POST-RENAL PROTEINURIA

- Intravascular hemolysis - Lower UTI/inflammation -
- Muscle injury Prostate fluid
- Inflammation and infection - Injury/Trauma - Vaginal
- Multiple Myeloma = proliferation of secretions
immunoglobulin producing plasma cells - Menstrual contamination
BENCE JONES PROTEIN – identified in serum
electrophoresis REACTION INTERFERENCES
*in urine: precipitates at 40-60 C (Cloudy) False (+)
- Highly alkaline urine interferes w/ the acid buffer
disoslves at 100 C (color change unrelated to CHON)
- Long contact of urine to the reagent pad
B. RENAL PROTEINURIA – actual renal damage - Contamination w/ quarternary ammonia
a. GLOMERULAR PROTEINURIA compounds, detergent and antiseptics
- High specific gravity
- Impaired selective filtration due to glomerular
False (-)
damage causing high molecular weight (and - Presence of non-albumin protein because the test is
negatively charged) substances to escape sensitive to albumin
through.
CAUSES: OTHER TEST FOR PROTEINS:
- Amyloidosis - Immune Complexes SSA (Sulfosalicylic Acid) Precipitation Test
- Toxic substances (SLE and Strep infect) - Cold precipitation that reacts equally on
all forms of proteins
*DIABETIC NEPHROPATHY 3mL of 3% SSA + 3mL urine
o May lead to renal failure CHON
o Indicated by MICROALBUMINURIA (which is Grading Description Range
(mg/dL)
not detected by rgnt strip)
Negative No increased turbidity <6
ALBUMIN EXCRETION RATE (AER) = expressed in Trace Noticeable turbidity 6 -30
ug/min or mg/24 hours. 1+ Distinct turbidity 30 – 100
o NORMAL AER = 0-20 ug/min 2+ Turbidity+granulation 100 – 200
o Microalbuminuria = 20-200ug/min 3+ Turbidity+gran+flocculation 200 – 400
(30-300mg/24hours) 4+ Clumps of Protein > 400
o Clinical Albuminuria = >200ug/min
MICRAL TEST – for detection of microalbuminuria IV. GLUCOSE – most frequently tested
- Strip employing Ab-enzyme conjugate that binds - Renal threshold is 160 – 180 mg/dL
albumin (Enzyme Immunoassay) Principle: Double sequential enzymatic reaction
(-) White (+) Red
Reagent: COPPER REDUCTION TEST (CLINITEST/BENEDICT’s

MULTISTIX: Glu Oxidase, Peroxidase, K Iodide TEST)
(Blue to green to brown) - non-specific test for reducing sugars
CHEMSTRIP: Glu Oxidase, Peroxidase, Principle: Copper reduction in the presence of
Tetramethylbenzidine (Yellow to Green) heat and alkali
Other Chromogens: aminopropylcarbazole (yelloworange- PROCEDURE:
brown) 5 gtts urine + 10 gtts Water + Clinitest Tablet
Ortho-toluidine (pink to purple)
Reaction interference:
False (+): Contamination with strong oxidizers and non- CLINITEST TABLET CONTAINS: CuSO4, NaCO3, Na
reducing sugars Citrate and NaOH
False (-): contamination/presence of reducing substances
sucas Vit. C
Pass-Through Phenomenon: > 2g/dL is present
OTHER SUGARS THAT COULD BE PRESENT IN URINE **Blue to Green to Yellow to Brick Red BACK TO
1. FRUCTOSE (Levulose) – Increased fruits, honey or BLUE.
syrup intake To prevent, use 2 gtts of urine.
2. GALACTOSE – Increased in infants with
galactosemia Glu Oxi Clinitest Interpretation
3. LACTOSE (Glu+Gal) – Increased in pregnancy, 1+ Neg Small amt of Glu
lactation and strict milk diet. Oxidizing agent interfering rgnt
4. PENTOSE (Xylulose) – Increased in benign 4+ Neg
strip
pentosuria Negative (+) Non glucose reducing substance
5. SUCROSE (Glu+Fru) – non reducing sugar. False (+)
in reagent strips but (-) in Copper reduction test.
Increased in intestinal disorders.
V. KETONES – results from increased FAT
METABOLISM due to inability to metabolize
CLINICAL SIGNIFICANCE OF GLUCOSURIA: carbohydrates.
HYPERGLYCEMIA RENAL DSO Principle: Legal’s Test (Sodium Nitroprusside
ASSOC. ASSOCIATED reaction)
Blood Glucose: Normal KETONE BODIES:
Increased 78 % -hydroxybutyric acid = major ketone BUT
Urine Glucose: Increased NOT DETECTED IN REAGENT STRIP.
Increased 20% Aceto-Acetic Acid = parent ketone
CAUSES: CAUSES: 2% Acetone = detected only when GLYCINE is
DM - impaired added.
Cushing Syndrome tubular
Pheochromocytoma reabsorption CAUSES:
Acromegaly such as in: - Diabetic Mellitus - Starvation
Hyperthyroidism FANCONI SYNDROME - Vomiting - Malabsorption
ACETEST TABLETS
TEST Hemoglobin Myoglobin Contains: Sodium Nitroprusside
Red/Pink Plasma Pale yellow
Sodium Phosphate
1. Plasma Test Decreased Increased CK and Lactose – for better color differentiation
haptoglobin Aldolase
2. Blondheim’s Test Precipitated Not precipitated - Can also be used for other body fluids
(Ammonium Sulfate) (-) (+) - Hygroscopic, easily absorbs moisture
- If the spx is not absorbed, use another
tablet
VI. BLOOD - (+) Tan or Pink or Violet

Principle: Pseudoperoxidase activity of Hgb - Detects 0.40mg/dL of Bilirubin in Urine
Reagents:
MULTISTIX: Diisopropylbenzene CLINICAL SIGNIFICANCE:
dehydroperoxidase tetramethylbenzidine - Hepatitis
CHEMSTRIP: Dimethyldihydroperoxyhexane - Biliary Obstruction
tetramethylbenzidine - Cirrhosis
(+) results: Uniformly green/blue = Hemoglobin/Myoglobin ICTOTEST (TABLETS)

Speckled/Spotted = Hematuria
 Less subject to interference and is
CLINICAL SIGNIFICANCE sensitive to 0.5 to 0.10 mg/dL
• Hematuria – presence of intact RBC CONTAINS:
• Hemoglobinuria – presence of  p-nitrobenzene-diazonium p-
hemoglobin pigment from RBC toluenesulfonate
destruction.  SSA
• Amounts of > 5 cells/microliter =  Sodium carbonate
significant  Boric acid
• Red colored urine needs confirmation in (+) Blue to purple color
the microscopic exam to confirm presence
of blood PROCEDURE:
10 gtts urine + Ictotest tablet + 1 gtt Water
MYOGLOBIN vs. HEMOGLOBIN After 5 secs, add another 1gtt water
HEMOGLOBINU MYOGLOBINU Read result after 60 seconds.
HEMATURIA
RIA RIA
Cloudy red Urine Clear Red Urine Clear Red Urine VIII. UROBILINOGEN - Bile pigment from
Seen In: hemoglobin degradation
- - IV hemolysis - Rhabdomyolysis  Present in small amount, <1 mg/dL
Glomerulonephr - Hemolytic anemia  Specimen of choice: afternoon urine
itis - Transfusion
- Renal Calculi reactions
(2pm to 4pm due to alkaline tide)
Principle: Ehrlich’s Reaction
Reagent:
BLONDHEIM’s TEST MULTISTIX: p-dimethylaminobenzaldehyde
5ml of spun urine + 2.8 g of NH3SO4 (80% CHEMSTRIP: 4-methoxybenzene-diazonium-
standard) tetrafluoroborate
- Stand for 5 mins
- Centrifuge or filter the soln. NOTE: Ehrlich reactive compounds-
- The supernatant is then examined for Porphobilinogen, Indican, p-aminosalicylic acid,
Blood rgnt strip. sulfonamides, methyldopa, procaine and
(+) Green to Blue ( - ) Yellow chlorprozamine
VII. BILIRUBIN - Presence in urine provides early CLINICAL SIGNIFICANCE:

indicaiton of liver disease. Detected long URINE URINE
CONDITION
before presentation of jaundice. BILIRUBIN UROBILINOGEN
- Conjugated bilirubin, water soluble Hemolytic
- Amber urine with yellow foam Disease of Neg +++
Newborn
Principle: Diazo Reaction
Liver Damage +/- ++
Reagents:
Bile Duct Neg/ Decreased
MULTISTIX: 2,4-dichloroaniline diazonium salt +++
Obstruction (normal strip)
CHEMSTRIP: 2,6-dichlorobenzene diazonium salt
WATSON-SCHWARTZ TEST Reagents:

 Differentiation of urobilinogen, MULTISTIX: p-arsanilic acid, tetrahydrobenzo-
porphobilinogen and other Ehrlich- quinolin-3-ol
Reactive Compounds. CHEMSTRIP: sulfanilamide, hydroxytetrahydro
 Extraction with CHLOROFORM and benzoquinoline
BUTANOL.
PROCEDURE: NOTES:
TUBE 1: 2mL urine + 2mL Chloroform + 4mL Na o Pink spots/edges is considered as negative
Acetate o Uniform pink is considered positive
TUBE 2: 2mL urine + 2mL Butanol + 4mL Na o (+) nitrite corresponds to 100,000
Acetate organisms/mL
Results: X. LEUKOCYTES
In presence of UROBILINOGEN  Significance: UTI/Inflammation, screen
for urine culture specimens.
Principle: Leukocyte Esterase

Reagents: Indoxylcarbonic acid ester
NOTES:
o WBC with Esterases: Neutrophil, Eosinophils,
In presence of PORPHOBILINOGEN Basophils, Monocytes
o Cells with Esterase: Histiocytes and
Trichomonas
o No esterase: Lymphocytes
o Strip can detect even lysed WBCs.
XI. ASCORBIC ACID

In presence of other Ehrlich Reactive compounds  Reducing agent
 Causes false negative reactions to: “BB
LNG”
Blood Nitrite
Bilirubin Glucose
Leukocyte
11th reagent pad: Vit C, Phosphomolybdate
HOESCH TEST (INVERSE EHRLICH REACTION)  (+) Molybdenum blue
 Rapid screening test for PBG (>2mg/dL)
Reagent: Ehrlich’s rgnt in 6M or 6N HCl MICROSCOPIC ANALYSIS OF URINE
 2 drops of urine + 2 mL of Hoesch Rgnt URINE SEDIMENT PREPARATION
 (+) Red 10 to 15 mL of Urine (ave. of 12mL)

IX. NITRITE - Rapid sreening test for Spin at 400 RCF for 5 minutes
UTI/Bacteuria 
 Considered as a valuable test for Decant urine (0.5 or 1mL remains)
detecting initial bladder infection (cystitis) 
 Performed in parallel with Leukocyte Transfer uL (0.02mL) sediment to glass slide with
esterase to determine the necessity for 22x22 coverslip
urine culture 
 SPECIMEN: First morning or 4hour urine Examine microscopically 10 LPF, 10 HPF under
Principle: Greiss Reaction reduced light
ADDIS COUNT – quantitative measure of formed MUST READ!!!

elements of urine using hemacytometer. THE ABOVE INFORMATION ABOUT THE TOPIC
o Specimen: 12-hour urine
CHEMICAL EXAMINATION OF URINE IS FROM
o Preservative: Formalin
Normal Values: THE FILE CM Review. Urinalysis and Renal
 RBC = 0 to 500,000/12hr Disorders.docx sent by Sir Rojohn.
 WBC = 0 to 1,200,000/12hr
 Hyaline Casts = 0 to 5,000/12 hr
READ AND REVIEW THE PROFESSOR’S
POWERPOINT IN CANVAS FOR COMPLETE
MICROSCOPY TECHNIQUES INFORMATION AND UNDERSTANDING OF THE
Bright Field Routine microscopy (low TOPIC.
refraction)
https://olfu.instructure.com/courses/131029/files/236
Phase- Enhanced visualization of highly
99959?module_item_id=14108749
Contrast refractile elements
Polarizing ID of chole in oval fat bodies, fatty
casts and crystals (maltese cross) Disclaimer: This trans may contain incomplete
Dark-Field ID of T. pallidum information about the topics. Use at your own risk.
Fluorescence Visualization of fluorescent Hehe
organism/substances
Interference 3D microscopy. And layer-by-layer
a.Normarsky imaging
b. Hoffman Differential/Bright field
Modulation/Birght field

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OUR LADY OF FATIMA UNIVERSITY

MEDICAL LABORATORY SCIENCE BATCH 2024

COURSE OUTLINE: PRELIMS QUALITY CONTROL

 Hippocrates - “uroscopy” 5th Century  Observe universal precaution

 1140AD – color charts was developed (20 color)

 Purpose of preservatives • Formalin 40% - 1 drop/ 30 mL of urine; is an excellent

Gauze-pad method- a gauze pad is used to collect the

• Thymol crystals – preserves glucose and sediments

KINDS OF URINE SAMPLE/ SPECIMEN

2. TIMED URINE SPECIMEN A. Glomerulus

URINE FORMATION • Passive Transport- movement of molecules across a

1. Renal Blood Flow membrane as a result of differences in their

content • Preservation - refrigeration, freezing, chemical

 Renin production = low plasma pressure and DRUG SPECIMEN COLLECTION

Nitrofurantion(Furadantin) Brown Yellow • Intravenous saline or glucose solution

Rifampin Bright Orange Red than1.018 at night

Riboflavin Bright Yellow Oliguria

METHODS: Harmonic Oscillation Densitometry

1. Urinometry - uses urinometer or hydrometer • Sound wave frequency

• Principle : water displacement or buoyancy • Automated instruments

• Disadvantage: requires 10-15ml

gravity of • Timing the fall of a drop of body fluid of known size,

• Distilled water: 1.001 through a definite distance in a mixture

• Potassium Sulfate: 1.015 • Heavier drop will fall faster

• Specimen cold- subtract

Reading is affected by: solute per kilogram of solvent

Refractometer maybe calibrated using the ff:

CHEMICAL EXAMINATION OF PARAMETERS OF DIPSTICK TESTING:

Sample problem: II. pH – important indicator for identification of

C. HARMONIC OSCILLATION DENSITOMETRY CAUSES OF BASIC URINE:

4. Proteins for prostatic discharge/vaginal **ORTHOSTATIC PROTEINURIA – due to increased pressure

CAUSES: C. POST-RENAL PROTEINURIA

Reagent: COPPER REDUCTION TEST (CLINITEST/BENEDICT’s

VI. BLOOD - (+) Tan or Pink or Violet

(+) results: Uniformly green/blue = Hemoglobin/Myoglobin ICTOTEST (TABLETS)

VII. BILIRUBIN - Presence in urine provides early CLINICAL SIGNIFICANCE:

WATSON-SCHWARTZ TEST Reagents:

Principle: Leukocyte Esterase

XI. ASCORBIC ACID

ADDIS COUNT – quantitative measure of formed MUST READ!!!

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