AUBF PRELIMS

Analysis of Urine & Body Fluids o You must monitor each test with
Safety in the Clinical Laboratory standards and controls (at least two
Renal Function levels)
Introduction to Urinalysis o New tests must be validated
Physical Examination of Urine Participating in Proficiency Testing
o External proficiency testing is mandated by
LABORATORY STANDARDS CLIA 88
CLSI (Clinical Laboratory Standards Institute) o Unknowns from CAP or other approved lab
Formerly NCCLS o Internal proficiency testing is also helpful for
Publishes national and international standard quality
documents on a variety of laboratory testing o Establishing a Quality Assessment
procedures and policies for guidance on Program Ensures
achieving better testing outcomes.
SAFETY IN THE CLINICAL LABORATORY
QUALITY ASSESSMENT
Variables Affecting the Quality of Laboratory

OSHA (Occupational Safety and Health

Administration)
Worker safety issues
CDC (Centers for Disease Control and Prevention)
Implements public health regulations, monitors
reportable diseases and trends, categorizes lab
tests

BIOLOGICAL HAZARDS
1. Chain of Infection
Source
Testing Mode of Transmission
Establishing a Quality Assessment Program Susceptible Host
o effective communication 2. Proper Handwashing and Personal Protective
o adherence to established policies Equipment (PPE)
o accurate and thorough documentation
practices Universal Precautions
o use of delta checks 1. All patients considered as possible carriers of
o timely verbal reporting of all critical
bloodborne pathogens.
values
2. Recommends wearing of gloves, face shields,
Performing Quality Control Tests
proper disposal of sharps.
o Quality Control
3. EXCLUDED: urine & body fluids not visibly
o A set of procedures and practices to
contaminated with blood
monitor the testing process and verify
the reliability, accuracy, and precision of
Body Substance Isolation
the test.
1. Modification from Universal Precaution
AUBF PRELIMS
2. Not limited to bloodborne pathogens Do not neutralize chemicals
3. Personnel at all times should wear gloves Chemical Handling
4. MAJOR DISADVANTAGE: does not Never mixed chemicals unless specified
recommend handwashing after removal of Wear goggles when preparing reagents
gloves unless visual contamination is present Do not pipette by mouth
Chemical Labeling
Standard Precautions Poisonous
P atient-care equipment Corrosive
H andwashing Carcinogenic

L inens
E nvironmental Control RADIOACTIVE HAZARDS

G loves Effects of radiation is cumulative

Degree of exposure related to:
o Time
o Distance
Personal Protective Equipment
o Shielding
1. Gloves
Wear measuring device to detect amount of
2. Fluid-resistant gowns
accumulated radiation
3. Eye and Face shields Post radioactive symbols on doors if so.
4. Centrifuge using capped tubes or cover
centrifuge when using ELECTRICAL HAZARDS
5. Special precautions to specimen submitted via Continually observe for frayed cords and
suspicious containers overloaded circuits
Handwashing Avoid unplugging equipments with wet hands or
- Hand contact as number one method of while cleaning.
infection transmission Remove source of electricity if electrical shocks
Disposal of Biological Waste are encountered or avoid transfer of the current
1. Incineration
2. Autoclaving FIRE/EXPLOSIVE HAZARDS R escue
3. Pick-up by certified hazardous waste company Store flammable A larm

chemicals properly C ontain

SHARP HAZARDS Compressed gas should E xtinguish
- Discard sharps in puncture resistant containers be located away from heat
located within work area. Fire blankets must be present in the lab
If persons clothes are burning, wrap in blanket
CHEMICAL HAZARDS to smother the flames
Chemical Spills PHYSICAL HAZARDS
Flush area with Avoid running in rooms and hallways
M ask, eye protection, face
large amount of Watch for wet floors
water shields
Bend knees when lifting heavy objects
Contaminated P atient Placement
Keep long hair pulled back
clothing should O ccupational Health & Blood
Avoid dangling jewelry
be removed Borne Pathogens
Maintain clean and organized work area
immediately G owns
Wear closed toe-shoes
AUBF PRELIMS
Renal pelvis
- Are large collection reservoir that joins calyces
RENAL FUNCTION together
Renal Physiology - It narrows as it exits the hilum to become the
Renal Blood Flow ureter.
Glomerular Filtration Renal Blood Flow
Tubular Reabsorption
Tubular Secretion
Renal Function Tests
o Glomerular Filtration Tests
o Tubular Reabsorption Tests
o Tubular Secretion & Renal Blood Flow
Tests

Parts and Function of the Urinary system

Kidney Each kidney having 1-2M nephrons

-bean shape organ that is roughly oval with a medial o Cortical nephrons
indentation. o Juxtamedullary nephrons
-Located on either side of the vertebral column and Renal arteries supplies blood to the kidney
Receives 25% of the blood pumped through the
extend from the level of the last thoracic vertebra to
heart at all times.
just above the third lumbar vertebra.
Renal blood flow = 1,200 ml/min
-Average size is 11cm x 7cm x 3cm. The left kidney is
Total Renal plasma flow= 600-700 ml/min
slightly larger than the right
-The right kidney is a little bit lower than the left, based on BSA 1.73 m
Blood enters the capillaries of the nephron:
because the liver takes some of the space above the
1. Afferent arteriole
right kidney
2. Glomerulus
-A heavy cushion of fat encases each kidney and holds it
3. Efferent arteriole
in position including connective tissue and renal 4. peritubular capillaries
fasciae. a. PCT - immediate reabsorption of
essential substances from the fluid
Hilum - A concave notch in the middle surface of the b. Loop of Henle - major exchanges of
kidney water and salt takes place between the
- Structure enter or leave the kidney through this blood and medullary interstitium.
notch. Thus, maintaining osmotic gradient
Renal cortex - The outer region (salt conc.) necessary for renal conc.
Renal medulla - The inner region c. DCT - final adjustment of urinary

Renal pyramids composition

5. vasa recta in the juxtamedullary nephron.
- A dozen or so triangular wedges that makes up
a. ALoH
much of the medullary tissue
b. DLoH
- The base of each pyramids faces outward and
the narrow papilla of each faces toward the
Renal physiology
hilum.
Calyx Cup-like structure that adjoins with the renal
papilla.
AUBF PRELIMS

a. Cellular Structure of the Capillary Wall

Capillary Wall Membrane
Basement Membrane (Basal Lamina)
Visceral Epithelium of the Bowmans Capsule
b. Hydrostatic & Oncotic Pressure
Hydrostatic Pressure in the arterioles
Oncotic Pressures in the Bowmans
Capsule
c. Renin-Angiotensin-Aldosterone System
Tubular Reabsorption
Active Transport
Movement of a substance across a cell membrane
and against an osmotic gradient
Glomerular Filtration Needs a carrier protein to transport substance;
1. Consists of 8 Requires energy
capillary lobes or Passive Transport PCT
tufts Movement of molecules o Glucose
across a membrane as o Amino acid
2. Glomerulus located
ALoH - salts in Chloride
within the Bowmans result of differences in the
DCT - Sodium
capsule electrical potential on
()Tubular Reabsorption
3. Nonselective filter of opposite sides of the
plasma substances membrane
Active Transport distinguishes excess solute
with molecular Does not need carrier protein
Does not require energy filtration and extent renal tubular damage
weight <70,000 da
PCT Water, urea Maximal Reabsorptive Capacity (Tm)
4. Factors influencing
ALoH urea, sodium - Highest level a substance is reabsorbed
the plasma filtration
DLoH & Collecting tubules before appearance of substance in the urine
process
Water
 AUBF PRELIMS
ex. Glucose is at 350 mg/min.
Renal threshold
- Plasma concentration at which active
transport stops
* ex. Glucose is at 160-180mg/dL
Countercurrent Mechanism
a. Selective reabsorption process
b. Prevents excessive water reabsorption
through the water-impermeable walls of the
ALoH
Renal Function Tests
Arginine Vasopressin or Antidiuretic Hormone Glomerular Filtration Tests
1. standard test measuring:
Decreased Decreased body
urine volume
Increased ADH
hydration a. Filtering capacity of the glomeruli
b. Measures rate at which kidneys are able to

Tubular Secretion remove filterable substances from the blood

Involves passage of substances from the blood 2. To test accuracy of the substance being measured:

in the peritubular capillaries to the tubular a. Substance analyzed must be neither

filtrate reabsorbed nor secreted by the tubules

Functions: b. Stable in a 24-hour collected urine

a. Elimination of waste products not filtered c. Consistent in the plasma level

by the glomerulus d. Availability of the substance in the body;

b. Regulation of acid base balance in the body availability of the tests for chemical

through secretion of hydrogen ions analysis of the substance

Tubular Secretion Clearance Test Substance

1. Urea
- Present in all urine specimen but replaced by
other substances
2. Inulin
- Polymer of fructose
- Extremely stable substance not reabsorbed nor
secreted by the tubules
- Not a normal body constituent and infused at a
constant rate throughout the testing period
- Seldom choice if suitable substance is already
present in the body
3. Radionucleotides
- Measures plasma disappearance of infused
substances thus eliminating need for
urine collection
- Injection of radionucleotides to:
o visualize plasma disappearance
radioactive material
o enable visualization of the filtration of
one or both kidneys
AUBF PRELIMS
4. 2microglobulin Standard Formula to calculate milliliters of plasma
- Dissociates from human leukocyte antigen and cleared per minute (C) is:
removed from the plasma by glomerular UV
filtration C=
P
5. Creatinine Urine creatinine (U) = 400 mg/dL
- Waste product of muscle metabolism and found Plasma creatinine (P) = 5.0 mg/dL
at a constant rate in the blood
Urine volume (V) = of 2500 mL, from a 24-hour
6. Cystatin C
specimen
- Low molecular weight
Glomerular filtration rate = ?
- Potential marker for long-term monitoring of
Calculation:
renal function
2500 mL mL
- Constant in serum levels V= =1.74
- Independent of age, gender, and muscle mass
60 minutes minute
(24 hours )
- Higher analysis cost 1 hour
mg mL
400 x 1.74
dL minute mL
C= =139.2
5.0 mg/dL minute
* Creatinine filtration and excretion
Calculations:
Clinical Significance in the interpretation of results for
1. Greatest source of error: improper timed
Creatinine Clearance:
specimen
1. Determines the number of functioning nephrons
2. GFR reported in mL/min.
2. Functional capacity of the nephrons
3. UV
P
Results may be used to:
U = urine creatinine in mg/dL
1. Measure extent of nephron damage
V = urine volume in mL/min.
2. Monitor effectiveness of treatment in the
P = plasma creatinine in mg/dL
prevention of further nephron damage
3. Determine feasibility of administering
Calculate the urine volume for a 4-hour specimen
measuring 1L: Cockcroft and Gault Formula:
4 hours X 60 minutes=240 minutes 140 age
C=
1000 mL mL mg
=4.17 serum creatinine ( )
240 minutes min dL

Tubular Reabsorption Tests

Also known as Concentration tests
Involves Water Deprivation Tests
a. Fishberg Test - patients deprived of
fluid for 24 hours prior to measuring the
specific gravity
b. Mosenthal Test compared the
volume of day and night urine samples
to evaluate concentrating ability
Free Water Clearance
AUBF PRELIMS
o How much water must be cleared each Urea and Creatinine: tests to differentiate urine
minute to produce a urine with the from other body fluids.
same osmolarity as with the plasma Urine Volume
o Determines the ability of the kidney to Depends on amount of water excreted by the
respond to the state of body hydration kidneys
Factors influencing its
Tubular Secretion and Diabetes mellitus Diabetes Insipidus
volume:
Definition Malfunctioned Decrease in production
Renal Blood Flow Tests of the pancreas/malfunctio or function of ADH
o Fluid intake
Disease ned insulin
1. Test to measure Urine increased Decreased to normal o Fluid loss from
Specific
tubular secretion Gravity nonrenal sources
of non- o Variations in the secretions of ADH
2. filtered substances and renal blood flow o Necessity to excrete increased amounts of

3. Test: -amino hippuric acid test (PAH) using the dissolved solids such as glucose and salts
Terms related to abnormal urine volume:
dye phenolsulfonphthalein
a. Polyuria excretion of increased
amount of urine at daytime
Titratable Acidity & Urinary Ammonia
b. Nocturia excretion of increased
1. Ability of kidney to produce acid urine depends
amount of urine at nightime
on tubular excretion of ammonia by the cells of c. Oliguria excretions of decreased
the DCT
amount of urine
2. Inability to produce acid urine: Renal Tubular d. Anuria cessation of urine flow
Acidosis Difference between Diabetes Mellitus from
3. Measurement of total hydrogen ion excretion in Diabetes Insipidus
urine
Specimen Collection
INTRODUCTION TO URINALYSIS 1. Clean, dry, leak-proof
Urine Formation 2. Should be disposable and screw-capped
kidneys continuously form urine as an 3. Wide mouth with flat bottom
utrafiltrate of plasma 4. Recommends 50mL capacity for:
Urine Composition a. Additional specimen for repeat collection
Urea (major dissolved solid) b. Enough room for swirling urine
Creatinine, uric acid
Inorganic substances: chloride, sodium, Specimen Handling
potassium in vivo and in vitro changes in urine
Others: sulfates and phosphates
Factors influencing its concentration: Specimen Integrity
a. Dietary intake After collection, urine delivered to lab promptly and
b. Physical activity tested within 2 hours
c. Body metabolism
d. Endocrine functions Specimen Preservation
e. Body positions Recommended:
May also contain cells, casts, crystals, mucus Refrigeration at 20-80C or at 40-60C
and bacteria Increases specific gravity when measuring
using Urinometer
AUBF PRELIMS
Ideal preservatives should be: Urin
a. Bactericidal e
b. Inhibit urease
c. Preserve formed elements in the sediment

collected under sterile condiitons

Types of Urine Specimens
Bacterial culture
1. RANDOM URINE SPECIMEN Specimens for left and right kidneys may be
Most commonly received
collected separately
May produce erroneous results caused by
dietary intake or physical activity prior to 8. MIDSTREAM CLEAN-CATCH URINE SPECIMEN
collection Alternative to catheterized specimen
Safer, less traumatic
2. FIRST MORNING URINE SPECIMEN Specimen less contaminated by epithelial cells
Ideal screening test and bacteriad
Essential for preventing false-negative Strong bacterial agents such as
pregnancy results hexachlorophene and povidone-iodine should
For evaluation of Orthostatic Proteinuria
not be used prior to collection
Provides a concentrated urine assuring
detection of chemicals and formed elements
9. SUPRAPUBIC ASPIRATED URINE SPECIMEN
External introduction of needle to the bladder
3. FASTING URINE SPECIMEN
Sample for bacterial culture completely free of
(SECOND MORNING SPX)
Second voided specimen after a period of extraneous contamination
For cytological examination
fasting

4. 2-HOUR POST PRANDIAL URINE SPECIMEN 10. THREE-GLASS COLLECTED URINE SPECIMEN
Patients instructed to void shortly before For prostatic infection
consuming routine meal and to collect urine 2
hours after eating
Monitors insulin therapy in patients with DM

5. GLUCOSE TOLERANCE TEST URINE

SPECIMEN
Collected to correspond with blood specimens
Tested for glucose and ketones
Tests the patients ability to metabolize 11. PEDIATRIC URINE SPECIMEN
measured amount of glucose and therefore Poses great challenge in urine collection
correlated with the renal threshold for glucose
12. URINE FOR DRUG TESTING
Chain of custody process that provides
6. 24-TIMED URINE SPECIMEN documentation for proper sample identification
Measures the kind and amount of chemicals from the time of collection to the receipt of
present laboratory results

7. CATHETERIZED URINE SPECIMEN

 AUBF PRELIMS
Physical Examination of Urine length of contact
COLOR
Varies from colorless to black RBC suspended in urine for hours will produce

Factors affecting variations in color: brown urine (oxidation of hemoglobin to

o Normal metabolic function methemoglobin)

o Physical activity brown urine may indicate glomerular bleeding
o Ingested materials Myoglobin & Hemoglobin
o Pathologic conditions - produces red urine and positive chemical test
Normal Urine Color: for blood
Dark yellow
Light yellow Hemoglobin breakdown of RED with red
Amber
Yellow plasma
Examine with good light source, against white Myoglobin breakdown of skeletal muscle
background with clear plasma
Urochrome
- pigment responsible for urine color
- established by Thudichum (1864)
- product of endogenous metabolism
produced at constant rate
- increased amounts in thyroid conditions,
fasting state, prolonged standing
Other pigments:
o Uroerythrin Non-pathogenic color of urine:
pink pigment evident in specimen after
menstrual contamination
refrigeration ingestion of highly pigmented food
attaches to urates producing pink color medications
o Urobilin ingestion of beets in genetically susceptible
imparts orange-brown color to urine that is not
individuals in alkaline urine
fresh Brown/Black
Dark Yellow
- If negative test for reagent strip for blood,
Amber
Abnormal Urine Color possible presence of melanin or homogentisic
Orange
Dark yellow/amber acid
may signify abnormal pigment Blue/Green
bilirubin (produces yellow foam) Pathogenic causes: bacterial infections and
Yellow-orange specimen intestinal tract infections resulting in increased
caused by administration of urinary indican
o phenazopyridine (pyridium Clorets
compounds) Purple staining in catheter bags:
o azo-gantrisin produces yellow foam o presence of indican or bacterial
when shaken and mistaken for infections associated with Klebsiella or
bilirubin Providencia
Red/Pink/Brown - Presence of blood
CLARITY
Factors affecting the changes of urine Refers to transparency or turbidity of a urine
in the presence of blood: specimen
amount of blood Common terminologies:
pH of urine o Clear no visible particles, transparent
AUBF PRELIMS
o Hazy few particulates, print easily Urine specific gravity of 1.025 measured from a
seen through urine specimen maintained at 140C, what is the corrected
o Cloudy many particulates, print
urine specific gravity?
blurred through urine 20 0 C14 0 C=6 0 C
o Turbid print cannot be seen through
1.0250.002
urine
1.023 corrected urine specific gravity
o Milky many precipitate or be clotted
Normal Clarity
Normally clear when freshly voided must be added for every 30C rise above the

Nonpathologic Turbidity Urine specific gravity of 1.025 measured from a

SEC on women resulting in hazy but normal specimen maintained at 260C, what is the corrected
urine urine specific gravity?
Specimen allowed to stand at room temp. 26 0 C20 0 C=6 0 C
Semen, fecal contamination, radiographic contrast 1.025+0.002
media, talcum powder, vaginal creams 1.027 corrected urine specific gravity
calibrated temp.
Pathologic Turbidity
RBC, WBC, bacteria commonly occuring Presence of protein and glucose in both urinometer
and refractometer:
Clarity of urine should always correspond to presence of protein : urine specific gravity
microscopic analysis usually rises by 0.003
presence of glucose: urine specific gravity
usually rises by 0.004

A urine specimen containing 5 g/dL of protein and 3

SPECIFIC GRAVITY g/dL of glucose has a specific gravity reading of
detects possible dehydration or abnormalities of 1.035. Calculate the corrected reading.
ADH g
density of a solution compared with the density of a
1.0350.015 5 (dL )
x 0.003 protein
similar volume of distilled water at a similar temp. 1.020 0.012( glucose)
influenced by the number of particles present and 1.008 corrected specific gravity
the sizes of the particles
aids in the evaluation of renal tubular function
Refractometer
Measures specific gravity through refractive
Urinometer or Hydrometer
index
Consists of a weighted float sinking at
Harmonic Oscillation Densitometry
o specific gravity of 1.000 in a distilled water
20 C calibrated temp. of urinometer
0 employs the principle of
For cold specimen, 0.001 must be subtracted from sound waves in Yellow Iris
the reading for every 30C that the specimen temp. is (International Remote
below the urinometer calibration temp. Imaging Systems)

Clinical Correlations
AUBF PRELIMS
specific gravity of plasma entering the Bacterial infection strong, unpleasant odor
glomerulus is usually 1.010 Diabetic ketones sweet or fruity odor
o Isosthenuria sg = 1.010 MSUD maple syrup
o Hyposthenuria below 1.010 Phenylketonuria Mousy
o Hypersthenuria above 1.010 Tyrosinemia Rancid
Isovaleric acidemia - Sweaty Feet
Methionine malabsorption Cabbage
ODOR
Contamination Bleach
Freshly voided urine faint aromatic odor
Breakdown of urea ammoniacal odor