AUBF (Macro, Chem, Micro) Flashcards
Included in PHYSICAL examination of urine
Color
Clarity
SG - now included in rgt strip
pigment present in urine that gives a yellow color (Thudichum)
Urochrome (normal pigment)
pink pigment, most evident in spx that have been REFRIGERATED, resulting in precipitation of AMORPHOUS URATES
uroerythrin
result of uroerythrin + urates in ACIDIC pH
pink urine sediment
oxidation product of UROBILINOGEN (normal urinary constituent), imparts an orange-brown (amber) color to urine that is NOT FRESH (longer standing)
Urobilin
pigment that may also be an indicator of hemolysis if extremely increased
checked: urobilinogen, bilirubin
urobilin
urine with amorphous urates in ACID pH
forms PINK precipitate
urine with amorphous phosphates in ALK pH
white precipitate
urine color:
oxidation of porphobilinogen to porphyrins
red
portwine
urine color:
heme biosynthesis problem —> porphyrias (enzyme deficiencies)
red
portwine
urine color:
melanoma
alkaptonuria
black
oxidation of melanogen (colorless pigment) to melanin (black pigment) - produced in excess
MELANOMA
condition with excreted BLACK urine
melanoma
(not alkaptonuria - only imparts black color during standing)
condition excreted as normal urine color and became BLACK during standing
alkaptonuria
homogentisic acid (metabolite phenylalanine) - imparts black color to ALK urine from person with inborn metabolism
alkaptonuria
metabolic problem in BABIES
inc pH (alk)
alkaptonuria
urine color:
pseudomonas inf
green
(pyoverdin)
urine color: clorets
blue-green
(pyocyanin)
what must be done before assessing urine clarity
MIX WELL
Reporting of urine clarity
Clear
Hazy
Cloudy
Turbid
Milky
Urine clarity:
no visible particulate
transparent
clear
Urine clarity:
FEW particulates,
print easily seen
hazy
Urine clarity:
MANY particulates,
print blurred
cloudy
Urine clarity:
print cannot be seen
turbid
Urine clarity:
may precipitate or be clotted (egg white-like) may be due to inc. lipid profile
milky
non-pathologic cause of urine TURBIDITY
squamous EC (contaminated collection)
mucus
amorphous phosphates, urates, carbonates
RCM
talcum powder
semen, spermatozoa
fecal contam
vaginal creams
pathologic cause of urine TURBIDITY
RBC
WBC
Bacteria
Yeast
Non-squamous EC (RTE cells)
Abnormal crystals
Lymph fluid (chyluria)
Lipids
differentiate hematuria from hemoglobinuria
hematuria: intact RBC microscopically
hemoglobinuria: hemolyzed RBC microscopically
eval of urine conc. (solutes present in urine)
SG
random urine sample SG
1.015-1.030
determine whether spx conc. is adequate to ensure chem test accuracy
SG
SG of water
1.000
SG of RCM
1.040
(differential to chole crystals)
SG: isosthenuric
1.010
SG: hyposthenuric
<1.010
SG: hypersthenuric
> 1.010
Current urine SG measurement methods
Refractometry
Osmolality
Reagent strip
principle of Refractometry
refractive index (read intersection betw. blue and white portion)
principle of Osmolality
changes in colligative properties by particle no.
principle of reagent strip
pKa changes of polyelectrolyte by ions present
Methods for manual SG measurement
Refractometer
Urinometer
measurement that requires temperature adjustment
urinometer
(5/9 ; F -32)
measurement that requires NO temperature adjustment
refractometer
calibration of manual SG measurement
subtract 0.003 for every 1 g of PROTEIN
subtract 0.004 for every 1 g of GLUCOSE
MACROSCOPIC screening for BLOOD seen microscopically
color
MACROSCOPIC screening for hematuria vs hemoglobinuria/myoglobinuria and pathologic and nonpathologic cause of turbidity seen microscopically
CLARITY
aka myoglobinuria
rhabdomyolysis
Hemoglobinuria vs. Hematuria vs. Myoglobinuria
Hemoglobinura: CLEAR, red
Hematuria: TURBID, red
Myoglobinuria: coca-cola like
MACROSCOPIC/CHEM SCREENING TEST for RBC, RBC casts seen microscopically
Blood
MACROSCOPIC/CHEM SCREENING TEST for casts and cells seen microscopically
protein
MACROSCOPIC/CHEM SCREENING TEST for bacteria and WBCs
nitrite
MACROSCOPIC/CHEM SCREENING TEST for WBCs, WBC cast, bacteria
Leukocyte esterase
(has the longest reading time - 120s)
MACROSCOPIC/CHEM SCREENING TEST for yeast seen microscopically
glucose
glucose in urine
glucosuria (ex. presence of yeast)
INCREASED
CHANGES IN UNPRESERVED URINE
“p(a)BaON”
- pH (alk)
- bacteria
- odor (ammoniacal)
- nitrite
DECREASED
CHANGES IN UNPRESERVED URINE
clarity
glucose
ketones
bilirubin
urobilinogen
RBCs, WBCs, Casts
Trichomonas
what happen to urine COLOR during long standing (unpreserved)? reason?
modified/darkened
Reason: oxidation or reduction of metabolites (urobilinogen —> urobilin “amber”)
what will happen to CLARITY during long standing of unpreserved urine?
reason?
turbid (decreased)
reason: bacterial growth and precipitation of amorphous materials
what will happen to GLUCOSE during standing in unpreserved urine? reason?
decreased
reason: glycolysis, bacterial use
what will happen to KETONES during standing in unpreserved urine? reason?
decreased (highly volatile, requires freshly voided urine for accurate measurement)
reason: volatilization, bacterial metabolism
what will happen to BILIRUBIN during standing in unpreserved urine? reason?
decreased
Reason: light exposure, photo oxidation to biliverdin
what will happen to UROBILINOGEN during standing in unpreserved urine? reason?
decreased
Reason: oxidation to urobilin “amber”
what will happen to RBC, WBC, casts during standing in unpreserved urine? reason?
decreased
Reason: disintegration in dilute ALK urine
what will happen to TRICHOMONAS during standing in unpreserved urine? reason?
decreased
Reason: loss of motility, death (if disintegrated, resembles WBC)
loss of motility and disintegration of trichomonas resembles?
WBCs
what will happen to ODOR during standing in unpreserved urine? reason?
increased (ammoniacal)
Reason: bacterial multiplication (urea —> ammonia)
what will happen to pH during standing in unpreserved urine? reason?
increased (alk)
reason: breakdown of urea to ammonia by urease-producing bacteria or CO2 loss
ex: alkaptonuria
what will happen to NITRITE during standing in unpreserved urine? reason?
increased
Reason:
- multiplication of NITRATE- reducing bacteria
- bacterial contamination
what will happen to BACTERIA during standing in unpreserved urine? reason?
increased
Reason: multiplication
cause of AROMATIC urine odor
NORMAL!!
cause of FOUL, AMMONIA-LIKE urine odor
Bacterial decomposition
UTI
cause of FRUITY SWEET urine odor
Ketones (Starvation, DM, Vomiting)
cause of MAPLE SYRUP urine odor
MSUD (inborn error of metabolism)
cause of MOUSY urine odor
Phenylketonuria (PKU)
cause of RANCID urine odor
tyrosinemia
cause of SWEATY FEET urine odor
isovaleric acidemia
cause of CABBAGE urine odor
Methionine malabsorption
cause of BLEACH urine odor
Contamination
cause of ROTTING FISH urine odor
trimethylaminuria
“galunggong-like” odor
Benedict’s test is only appropriate for?
glucose and other reducing sugars (except SUCROSE - nonreducing)
Principle of benedict’s test
Copper reduction
Benedict’s test reaction
Cupric ions (Cu2+) + reducing sugar —heat + alkali –> carboxylic acid + Cuprous oxide (Cu2O)
Cupric ions - blue solution
reducing sugar - in urine
Cuprous oxide - indicator that gives brick red ppt
ratio of benedict’s soln to urine
1:1
Negative (-) Benedict’s test
No color change, remains BLUE
Trace (+/-) Benedict’s test
Green opacity
No precipitate
+1 Benedict’s test
Green solution
Yellow precipitate
+2 Benedict’s test
Green to yellow solution
Yellow precipitate
+3 Benedict’s test
Muddy orange solution
Yellow precipitate
+4 Benedict’s test
Orange to brick red precipitate
Urine chem test for PROTEIN
Sulfosalicylic Acid precipitation (SSA) test
Principle of SSA
Precipitation of urine protein by STRONG acid
A cold precipitation test that reacts equally with all forms of protein
SSA test
Explain SSA test procedure
- 3 mL of 3% SSA reagent to 3 mL centrifuged urine (1:1)
- Mix by inversion and observe cloudiness/turbidity
- Grade the degree of turbidity
Protein measured if SSA test shows negative turbidity
<6 mg/dL
Protein measured if SSA test shows trace turbidity
6-30 mg/dL
Protein measured if SSA test shows 1+ turbidity
30-100 mg/dL
Protein measured if SSA test shows 2+ turbidity
100-200 mg/dL
Protein measured if SSA test shows 3+ turbidity
200-400 mg/dL
Protein measured if SSA test shows 4+ turbidity
> 400 mg/dL
Describe turbidity in SSA test if the protein is <6 mg/dL
NEGATIVE
No increase in turbidity
Describe turbidity in SSA test if the protein is 6-30 mg/dL
Trace (+/-)
Noticeable turbidity
Describe turbidity in SSA test if the protein is 30-100 mg/dL
1+
Distinct turbidity, NO granulation
Describe turbidity in SSA test if the protein is 100-200 mg/dL
2+
Turbidity, granulation, NO flocculation
Describe turbidity in SSA test if the protein is 200-400 mg/dL
3+
Turbidity, granulation, flocculation
Describe turbidity in SSA test if the protein is >400 mg/dL
4+
Clumps of protein
Classic test for differentiating UROBILINOGEN and PORPHOBILINOGEN
Watson-Schwartz Test
Reagents used in Watson-Schwartz test
Chloroform
Butanol
location of Chloroform in Watson-Schwartz test
BOTTOM
location of Butanol in Watson-Schwartz test
UPPER
Chloroform: Soluble
Butanol: Soluble
Urobilinogen
Chloroform: Insoluble
Butanol: Insoluble
Porphobilinogen
Chloroform: Insoluble
Butanol: SOLUBLE
Other Ehrlich reacting compounds
Watson-Schwartz test
Color if SOLUBLE
RED
Watson-Schwartz test
Color if INSOLUBLE
YELLOW
Analyte:
Heat & Acetic Acid
Nitric Acid test
Albumin/Protein
End result:
Heat & Acetic Acid
Nitric Acid test
Turbidity
Analyte:
Benedict’s test
Glucose and other reducing sugar
End result:
Benedict’s test
Brick-red precipitate
Analyte:
Heat Precipitation Test
Bence Jones Protein
End result:
Heat Precipitation test
Turbidity
Analyte:
Rothera
Ketones
End result:
Rothera
Red purple
Analyte:
Gunning
Ketones
End result:
Gunning
Iodoform crystals (microscopic)
Analyte:
Gerhadt
Ketones
End result:
Gerhadt
Bordeaux red
Analyte:
Seliwanoff
Levulose
End result:
Seliwanoff
Orange –> red
Analyte:
Rubner
Lactose
End result:
Rubner
Brick-red precipitate
Analyte:
Gmelin
Bile
End result:
Gmelin
Play of colors (green, red, yellow)
Analyte:
Smith
Bile
End result:
Smith
Green RING
Analyte:
Ehrlich
Urobilinogen
End result:
Ehrlich
Cherry red
Analyte:
Schlesinger
Urobilin
End result:
Schlesinger
Fluorescent GREEN
Analyte:
Obermayer
Indican
End result:
Obermayer
Indigo BLUE –> Indigo RED
Analyte:
Guaiac
Hemoglobin
End result:
Guaiac
Blue junction
Analyte:
Ammonium Sulfate
Hemoglobin
End result:
Ammonium Sulfate
HGB precipitation
Analyte:
Thormalen
Melanin
End result:
Thormalen
Prussian Blue
Analyte:
Sulkowitch
Calcium
End result:
Sulkowitch
Turbidity
Analyte:
Fantus
Chloride
End-result:
Fantus
White precipitate
Microalbumin testing (immunologic tests)
Micral test
Immunodip
Micral test principle
Enzyme Immunoassay
Sensitivity of Micral test
0-10 mg/dL
Reagents used in Micral test
Gold-labeled Ab
B-galactosidase
Chlorophenol red galactoside
Interference in Micral and Immunodip
FALSE NEGATIVE due to DILUTED URINE
ImmunoDip principle
Immunochromographics
Sensitivity of ImmunoDip
1.2 - 8.0 mg/dL
Reagents used in ImmunoDip
Ab-coated BLUE LATEX particles
Normal protein excreted by the body
<10 mg/dL
100 mg/24 hrs
Albumin level of 20-200 mg/L in urine indicating kidney damage
Microalbuminuria
Albumin level of >200 mg/L in urine
Albuminuria
Urine must be delivered and tested within ___ hours
2 hours
Causes precipitation of amorphous urates/phosphates, and other non-pathologic crystals in urine
MOST COMMON method of urine preservation
Refrigeration
This may cause some crystals to dissolve
Warming at 37C
Recommended type of urine spx
MIDSTREAM CLEAN CATCH URINE (morning, concentrated)
standard amount of urine
10-15 mL
frequently used urine vol in laboratory
12 mL
urine container capacity
50 mL
T/F
Report if urine vol is not adequate (<10 mL) but always check the medical diagnosis
TRUE
Daily urine output
1200-1500 mL (600-2000 mL)/day
decreased urine output
what are the volumes?
OLIGURIA
Infants: <1 mL/kg/hr
Children: <0.5 mL/kg/hr
Adult: <400 mL/day
cessation or failure of kidneys to produce urine
ANURIA
T/F
Kidneys excrete 2-3x more in day than night
T
Increased urine output at NIGHT
NOCTURIA
Increased urine output daily
What are the volumes?
POLYURIA
Children: 2.5-3.0 mL/kg/day
Adult: >2.5 L/day
Urine centrifugation
5 mins
400 RCF or 1,500 RPM
Volume of urine after removal of supernatant following centrifugation
0.5 - 1 mL
(uniform amt of urine and sediment)
Volume of sediment examined
20 uL (0.02 mL)
Purpose of examining urine sediment in LPO
view CASTS
ascertain the general composition of sediments
Purpose of examining urine sediments in HPO
Identification of urinary sediments
This method may affect casts reading wherein they have the tendency to locate near the edges of the cover slip. What is recommended to eliminate this?
Conventional Glass Slide method
Remedy: scanning of coverslip parameter
Point of reference in examining urine sediment
Epithelial cells (largest)
Reported as ave. no. per LPF
casts
Reported as ave. no. per 10 HPF
RBC
WBC
Reported as semiquantitative (rare, few, moderate, many / 1+, 2+, 3+, 4+)
EC
Crystals
Other sediments
Most frequently used stain in UA
Sternheimer-malbin stain
Components of Sternheimer-malbin stain
Crystal violet
Safranin O
Commercially available Sternheimer-malbin stain
Sedi stain
KOVA stain
Delineates structures and contrasting colors of the nucleus and cytoplasm
Sternheimer-malbin stain
used to confirm the presence of TAG, neutral fats, and chole
ID free fat droplets and lipid-containing cells and casts
Lipid stains (Oil red O, Sudan III) + Polarizing microscope
color of TAG and neutral fats using Lipids stains
orange-red
T/F
Oil red O and Sudan III cannot help visualize CHOLE alone. It requires polarizing microscope.
T
Used for ID of bacterial casts, which can be confused from granular casts
Gram stain
Components of Gram stain
Crystal violet (primary stain)
Iodine (mordant)
Alcohol (decolorizer)
Safranin (secondary stain)
Gram (+) bacteria color:
Gram (-) bacteria color:
Gram (+) bacteria color: purple/violet
Gram (-) bacteria color: pink/red
preferred stain for urinary EOSINOPHILS in cases of DRUG-INDUCED ALLERGIC RXN producing inflammation of the renal interstitium
Hansel stain
Components of Hansel stain
Methylene Blue
Eosin Y
Stain for Iron (hemosiderin granules: blue)
Stains positive during hemolysis and bleeding
Prussian blue stain
identifies YELLOW-BROWN granules of hemosiderin in cells and casts
Prussian blue stain
T/F
The type of microscopy used depends on the specimen type, refractive index, and ability to image unstained living cells
T
objects appear dark against a light BG
MOST FREQUENTLY USED in clin lab
Bright-field microscopy
type of microscopy that aids in Treponema pallidum ID
Dark-field microscopy
enhances visualization of elements with LOW refractive indices, such as:
HYALINE CASTS
MIXED CELLULAR CASTS
MUCUS THREADS
TRICHOMONAS
Phase-contrast microscopy
Aids in ID of cholesterol in oval fat bodies, fatty casts, and crystals
Polarizing microscopy
Allows visualization of naturally fluorescent microorg or those stained by a fluorescent dye including labeled Ag and Ab
Fluorescence microscopy
Produces a 3D microscopy image and layer by layer imaging
Interference contrast
Frequently performed independently of routine UA for detection of malignancies of the LOWER urinary tract
CYTODIAGNOSTIC URINE TESTING
Stain used in cytodiagnostic urine testing
papanicolaou stain (provides additional method for detecting and monitoring RENAL DISEASE)