CLINICAL MICROSCOPY Flashcards
Potential harmful microorganisms
BIOLOGICAL HAZARD
3 routes of infection
- Inhalation
- Ingestion
- Direct inoculation or skin contact
6 components (Chain of infection)
IREMES
1. Infectious agent
2. Reservoir
3. Exit portal
4. Mode of transmission
5. Entry portal
6. Susceptible host
PPE
- Gloves
- Fluid resistant gowns
- Eye and face shields
- Countertop shields
The primary method of infection transmission
HAND CONTACT
BEST WAY to break the chain of infection
HANDWASHING
HANDWASHING PROCEDURE
- Wet hands with warm water.
- Apply antimicrobial soap.
- Rub to form a lather, create friction, and loosen debris.
- Thoroughiy clean between fingers, including thumbs, under fingernails and rings, and up to the
When hands wrist for at least [15 (or 20) seconds] - Rinse hands in a DOWNWARD POSITION.
- Dry with a paper towel.
- Turn off faucets with a clean paper towel to apply alcohol prevent recontamination.
HANDWASHING SONG
Happy Birthday (2x)
When hands are visibly soiled
Wash hands with soap and water
When hands are NOT visibly soiled
Apply alocohol based hand rub
(ex. Sanitizer)
ALL biological waste, except _____, must be placed in appropriate containers labeled with the biohazard symbol.
URINE
Accepted BIOHAZARD label
FLUORESCENT ORANGE
Disinfection of the sink using a _____, should be perform _____. (Effective for ____; used for disinfecting countertops and spills)
1:5 or 1:10 dilution of sodium hypochlorite
Daily
1 month
Eliminates many or all pathogenic microorganisms, except bacterial spores.
DISINFECTION
Chemical spills best first aid
Flush the area with amounts of water for at least 15 minutes the seek for medical attention.
What will you do for alkali or acid burn in the eye?
Wash out eye thoroughly with running water for 15 minutes.
DO NOT NEUTRALIZE CHEMICALS that come in contact with the skin.
Acid spills on floors can be neutralized and then soaked up with wet rags or spills.
CHEMICAL HANDLING
ALWAYS ADD ACID TO WATER
NFPA number 1 quadrant
YELLOW
(Reactivity/Stability hazard)
May deteriorate/ detonate
4
2 represents?
Violent chemical change
3 represents
Shock & heat may deteriorate/detonate
0 and 1 represents
0- stable
1- unstable when heated
Second quadrant
WHITE quadrant (Specific hazard)
OXY- oxidizer
ACID- acid
ALK- alkali
COR- corrosive
W- use no water
Radiation
3rd quadrant
BLUE quadrant (HEALTH hazard)
0, 2 and 3 represents?
0- normal material
2- hazardous
3- extreme danger
1 and 4 represents
1- slightly hazardous
4- deadly
4th quadrant
RED (flammability hazard)
Will not burn
0
1 and 2 represents
1- above 200^F
2- below 200^F
3 and 4 represents
3- below 100^F
4- below 73^F
DEGREE OF HAZARDS (hazard index)
“No SMS Ex’s”
0- NO/Minimal hazard
1- Slight hazard
2- Moderate hazard
3- Serious hazard
4- Extreme/Severe
Electrical hazard
*DO NOT OPERATE equipment with wet hands.
*All electrical equipment is grounded in a 3-cronsed plug to avoid electric shock.
*If electrical shock occurs, never touch the person er the equipment involved.
- Turn off the circuit breaker
- Unplug the equipment
- Move the equipment using a nonconductive glass or wood object
All laboratory personnel must be involved in laboratory fire drills at least ____
ANNUALLY
When a fire is discovered?
RACE
RESCUE anyone in immediate danger
ALARM (Activate the institutional fire alarm system)
CONTAIN (Close all doors to potentially affected areas)
EXTINGUISH/ EVACUATE (Attempt to extinguish the fire, if possible; exit the area) ALWAYS CHOOSE EXTINGUISH!!
To operate a Fire extinguisher?
PASS
PULL the pin
AIM the base of the fire
SQUEEZE handles
SWEEP nozzle side to side
Ordinary combustibles; PAPER, cloth, rubbish, plastic, WOOD
Type of fire?
Type of extinguisher?
Type A fire
WATER, dry chemical, loaded steam
Cooking media: grease, oils, tats
Type of fire and extinguisher?
Type K fire
Liquid designed to prevent splashing and cool the fire
Flammable metals: mercury, magnesium, sodium, lithium
Type D fire
Metal X, sand; dry powder; fought by fire fighters only
Flammable liquids: grease, gasoline, paints, oil
Type B fire
Dry chemical, carbon dioxide, halon foam
Electrical equipment and motor switches
Type C fire
Dry chemical, carbon dioxide, halon
Detonation (Arsenal fire)
Type E fire
Allowed to burn out and nearby materials protected
Best fire extinguisher for electrical fire?
HALON (BC)
The most common all purpose extinguishers
ABC (dry chemical extinguishers)
Kidney weights approximately ____ & measures ____
150 g
12.5 cm x6 cm x2.5 cm
Ureter length ___
Urethra - ___ cm long in women and ___ cm long in men
About every ______, small amounts of urine are emptied into the bladder from the ureters.
Urine is actually a fur biopsy of the kidney
25 cm long
4 cm long in women
24 cm long in men
10-15 seconds
Basic strustural & functional unit of the kidney
Nephrons
1. Cortical nephrons (85%)
- removal of waste products and nutrients reabsorption
2. Juxtamedullary
- urine concentration
1 to 1.5 million nephrons PER kidney
Consists of glomerulus and renal tubules
ORDER OF URINE FORMATION:
- Glomerulus
- Proximal convoluted tubule (PCT) -NEAR
- Loop of Henle (LH)
- Distal convoluted tubule (DCT)- FAR
- Collecting duct (CD)- last part of nephron
- Calyx
- Renal Pelvis
The kidney receives ____ of the total cardiac output.
25%
ORDER of RENAL blood flow
- RENAL ARTERY
- blood in
-unfiltered - Afferent arteriole
- approaching - Glomerulus
- plasma filtration of substances - Efferent arteriole
- exiting - Peritubular capillaries
- capillaries surrounding renal tubules (reabsorption and secretion) - Vasa recta in LH (selective reabsorption)
- Renal vein
- blood out
- filtered
2 kidneys
Total renal blood flow:
1,200 mL/min
Total renal plasma flow:
600-700 mL/min
The “working portion” of the kidney
Coil of approx. 8 capillary lobes (capillary tuff) located w/in the Bowman’s capsule
★Attached to the glomerular basement membrane are the podocytes (epithelial cells)
★ Resembles a SIEVE “salaan”
Non-selective filter of plasma substances with MW of <70,000 Da - easy pass
Approximately 1% of the filtered plasma volume is actually excreted as urine
GLOMERULUS
GLOMERULAR FILTRATION BARRIER
- Capillary endothelium with its large open pores “ Fenestrated”
- Trilayer basement membrane (lamina rara interna, lamina densa, lamina rara externa)
- Filtration diaphragm found between the podocytes of Bowman’s space
What are the glomerular filtrate?
SWAGU
Salts
Water
Amino acids
Glucose
Urea
Glomerular filtrate SG
- 010
A protein that is not filtered by the glomerulus because of its negative charge.
Has a molecular weight of 69, 000 Da
Can be positive if the pH is _____
ALBUMIN
<4.9
First function to be affected in renal disease
TUBULAR REABSORPTION
RENAL THRESHOLD FOR GLUCOSE
160-180 mg/dL
Alter urine concentration
PCT, LH, DCT, CD
The major site for reabsorption
PCT (65%)
Highly impermeable to water
Ascending LH
During Hyponatremia what will happen to amino acids, glucose and urea?
Aldosterone _____ (increase or decrease)
ADH ______
100% amino acids are reabsorbs
Glucose reabsorbs (<RTG)
40% of urea will be reabsorbs.
Aldosterone- INCREASED
ADH - INCREASED
Hypernatremia
SWGU will be excreted in the urine.
Aldosterone -DECREASED
ADH- DECREASED
0% amino acids
More than or equal renal threshold for GLUCOSE
60% urea will be excreted
What are the substances that are ACTIVELY transport by the PCT?
Glucose
Amino acids
Salts
What is the substance that is ACTIVELY transport by the ASCENDING LH?
Chloride
What is the substances that is ACTIVELY transport by the PCT and DCT?
Sodium
What is/are the substances that is/are PASSIVELY transport by the PCT, ascending LH and CD?
WATER
What is/are the substances that is/are PASSIVELY transport by the PCT, ascending LH?
UREA
What is/are the substances that is/are PASSIVELY transport by the ascending loop of henle?
SODIUM
Regulates WATER REABSORPTION in the DCT and CD?
Anti-Diuretic hormone (ADH/VASOPRESSIN)
ADH deficiency
DIABETES INSIPIDUS
DI- “Dami Ihi”
ADH excess
Syndrome of inappropriate ADH secretion (SIADH)
Increased Body Hydration ___ ADH ____ Urine volume
Decreased, Increased
Decreased body hydration ____ADH ____urine volume
INCREASED, DECREASED
Regulates SODIUM REABSORPTION
ALDOSTERONE
RAAS is activated by:
Decreased Na, Decreased BP
Dilation: afferent
Constriction: efferent
Activated RAAS
Increased BP
Constriction: Afferent
Effects of Angiotension II
- Release of Aldosterone S ADH (Increased Sodium & water reabsorption)
- Vasoconstriction (Increased blood pressure)
- Corrects renal blood flow
Juxtaglomerular apparatus consists of the JG cells produces ____ in the afferent arteriole
RENIN
Macula Densa ________ of the DCT
Detects decrease in BP
ACTION OF RAAS
- Dilates the afferent arteriole & constricts the efferent arteriole
- Stimulates sodium reabsorption in the PCT
- Triggers the adrenal cortex to release aldosterone to cause sodium reabsorption & potassium excretion in the DCT and CD
- Triggers release of anti-diuretic hormone by the hypothalamus to stimulate water reabsorption in the CD
2 Major Functions of tubular secretion
- Regulation of the acid-base balance in the body through secretion of hydrogen ions (in the form of NH4 and H2P04).
- Elimination of waste products not filtered by the glomerulus
Major site for removal of non-filtered substances.
PROXIMAL CONVOLUTED TUBULE
Failure to produce an acid urine due to inability to secrete hydrogen ions
RENAL TUBULAR ACIDOSIS
What is the relationship between pH and Hydrogen ions?
INVERSELY PROPORTIONAL
RENAL TUBULAR ACIDOSIS
Increased hydrogen ions in the blood
Decreased hydrogen ions in the urine
Increased pH = alkaline
Used to evaluate glomerular filtration.
Measure the rate at which the kidneys are able to remove a filterable substance from the blood.
CLEARANCE TEST
What clearance test is obsolete, oldest, not accurate, present in all urine specimen (40% is reabsorbed)
UREA
Most common but not reliable indicator to patient suffering from muscle wasting diseases.
CREATININE (ENDOGENOUS)
GOLD STANDARD test for glomerular filtration
INULIN (EXOGENOUS)
better marker of renal tubular function than of GFR
Beta2-microglobulin (MW: 11,800 Da)
Creatinine Clearance Formula
Ccr (mL/min)= UV/P x 1.73m^2
Where:
Ccr= Creatinine clearance
U = Urine creatinine (mg/dL)
P= Plasma creatinine
V= Urine volume (mL/min)
A= Body surface area
Normal values
Male 107-139 mL/min
Female 87-107 mL/min
Is a measure of the completeness of a
24-hour urine collection
CREATININE CLEARANCE
By far the greatest source of error in any clearance procedure utilizing urine is the use of ______
IMPROPERLY TIMED URINE SPECIMENS
Around 7-10% of creatinine is secreted by the renal tubules.
Variables used in Estimated GFR formula developed by cockgroft and gault
Age
Sex
Body weight in kg
Formula:
Ccr= (140–age)(body wight in kg)/72x serum creatinine in mg/dL
Results x 0.85 (if female)
Modification of Diet in Renal disease (MDRD) system formula variables:
Ethinicity
BUN
Serum albumin
used to evaluate tubular reabsorption
CONCENTRATION TESTS
Patient is deprived of fluid for up to 24 hours
# Urine S.G. after 12-hour restricted fluid diet is about 1.022 or more
# Urine S.G. after 24-hour restricted fluid diet is about 1.026 or more
Obsolete test
- Fishberg test
Patient maintains normal diet and fluid intake
Compare day & night urine in terms of volume & S.G.
Obsolete test
- Mosenthal test
Influenced by the number & density of particles in a solution
Recently used tests
1. Specific gravity
Influenced by the number of particles in a solution
More preferred than S.G. determination
More precise than osmolarity because it does not vary with temperature Methods include freezing point osmometry & vapor pressure osmometry
NV =1-3x (275 to 900 mOsm/kg) than of serum (275 to 300 mOsm/ kg)
- Osmolality
most commonly used; reference method for tubular secretion and renal blood flow
p-aminohippuric acid (PAH) test
obsolete; results are hard to interpret
Phenolsulfonphthaiein (PSP) test
inferred diagnoses from urine evaluation
Hippocrates, Aristotle and the ancient Egyptians
4 temperaments/ humors:
- Sanguineous (blood)
- Choieric (vellow bile)
- Phlegmatic (phlegm)
- Melancholic (black bile)
The __________ of urine was used by the Babylonians and Egyptians to detect diabetes
Hindu physicians noticed that ________ attracted ants
“taste test”
“honey urine”
Uroscopy; first documented the importance of sputum examination
Hippocrates
Albuminuria by boiling urine
Frederik Dekkers
Wrote a book about “pisse prophets” (charlatans)
Thomas Bryant
Examination of urine sediment
Thomas Addis
Introduced urinalysis as part of doctor’s routine patient examination
Richard Bright
Urochrome
Ludwig Thudichum
Cerebrospinal fluid
Domenico Cotugno
Phenylketonuria
Ivan Folling
Alkaptonuria
Archibald Garrod
Cystine calculi
William Wollaston
Benedict’s reagent
Stanley Benedict
URINE COMPOSITION
_____% water
_____% solids (______ total solids in 24 hours)
95-97% water
3-5% solids (60% total solids in 24 hours)
Total solids
_____ grams organic
_____major organic compound
_____2nd organic compound
35 grams
UREA
CREATININE
_____grams inorganic compound
_____major inorganic compound >_____ >______
_____principal salt
25 grams
Chloride > Sodium > Potassium
NaCl
For routine and qualitative urinalysis
* Ideal for cytology studies (ONLY IF with prior hydration, & exercise 5 mins before collection!
- RANDOM/OCCASIONAL / SINGLE
Ideal specimen for routine urinalysis and pregnancy testing (hEG)
Often preferred for cytology studies/ cyto diagnostic urine testing
Most concentrated and most acidic - allows well preservation of cells and casts
For evaluation of orthostatic pruteinuria.
Patient voids before going to bed, and immediately on rising from sleep collects urine specimen
Alternative for cyto
FIRST MORNING
!!concentrated>hypertonic>shrink cells
Hypocentilation>respiratory acidosis
2nd voided urine after a period of fasting
For glucose determination
SECOND MORNING /FASTING
For diabetic screening or monitoring
Preferred for testing glucose
2-hour post-prandial
Optional with blood samples in glucose tolerance test
Glucose tolerance
At least 2 voided collection
Series of blood and urine samples are collected at specific time intervals to compare concentration of a substance in urine with its concentration in the blood
Used in the diagnosis of diabetes
Fractional specimen
For routine screening and bacterial culture
Patient should thoroughly cleanse his glans penis or her urethral meats before collection
Less contaminated by epithelial and bacterial cells
MIDSTREAM CLEAN CATCH
May be urethral or ureteral
For bacterial culture
CATHETERIZED
Abdominal wail is punctured, and urine is directly aspirated from the bladder
Bladder urine for anaerobic bacterial culture and urine cytology
Most sterile
SUPRAPUBIC ASPIRATION
Use of soft, clear plastic bag with adhesive
Sterile specimen obtained by catheterization or suprapubic aspiratica
Urine collected from diaper is NOT recommended for testing
PEDIATRIC SPECIMEN
For prostatic infection
THREE -GLASS TECHNIQUE
if the # of WBC and bacteria in the 3rd spx is 10x GREATER than that of the 1st
PROSTATITIS
Serves as a CONTROL for bladder & kidney infection.
2nd specimen
If control is (+) for WBCs and bacteria, the results from the 3rd specimen are considered invalid
If may laman <UTI></UTI>
The four-glass method consists of bacterial cultures of the
initial voided urine (VB1), midstream urine (VB2), expressed prostatic secretions (EPS), and a post-prostatic massage urine specimen (VB3). Urethral infection or inflammation is tested for by the VB1, and the VB2 tests for urinary bladder infection. The prostatic secretions are cultured and examined for white blood cells. Having more than 10 to 20 white blood cells per high-power field is considered abnormal.
STAMEY-MEARS TEST EOR PROSTATITIS
At start time, patient empties bladder into toilet; then all subsequent urine is collected At end time, patient empties bladder into collection container
Requires preservative - it depends on the test performed
24-hour (Ex: 8 AM -> 8 AM)
For addis count
12-hour (Ex: 8 AM -> 8 PM)
for nitrite determination
Urine remains in bladder for at least 4 hours before voiding
4-hour
(First morning urine- best for nitrite determination)
For UROBILINOGEN determination
Afternoon (2-4 PM)
Process providing documentation of proper sample ID from the time of collection to the receipt of laboratory results.
CHAIN OF CUSTODY
Required urine volume for DRUG test
30-45 ml
Drug test Container capacity
60 ml
Optimal temperature of urine in DRUG test
32.5- 37.7°C (checked within 4 minutes)
Added to the toilet water reservoir to prevent specimen adulteration
Blueing agent
Urine containers should have a wide base, and has an opening of at least ______ cm. The wide base prevents spillage, and a _____cm opening is an adequate target for urine collection.
4 cm
24-hr urine containers should hold up to _____ and may be colored to protect light sensitive analytes.
Addition of urine before the start of 24-hour collection period causes ________ results.
Failure to include urine at the end of 24-hour collection period causes ______ results
3L
False increased
False decreased
When both a routine UA and culture are requested on a catheterized or midstream collection, what will you do first?
CULTURE should be performed first to prevent contamination of the specimen
Following collection, urine specimens should be delivered to the laboratory promptly and tested within ________
(Strasinger, Harr); ideally within _____ minutes (Turgeon)
Physical, chemical and microscopic characteristics of a urine specimen begin to change _________
Within 2 hours
Ideally- 30 minutes
AS SOON AS THE URINE IS VOIDED
CHANGES IN UNPRESERVED URINE
Increased analytes?
Decreased analytes?
Increased analytes
-pH
-Bacteria
-Odor
-Nitrite
Darkened/Modified
-Color
Decreased analytes
Clarity
Glucose
Ketone
Blood
Urobilinogen
RBC/WBC/Casts
Trichomonas- resembles WBC
Least affected analyte in unpreserved urine?
PROTEIN
What is the ideal urine preservative?
Ideal urine preservative does not exist (Sirasinger)
Does not interfere with chemical tests
Raises SG by hydrometer
Precipitates amorphous phosphates and urates
Preservative of choice for routine UA & urine culture
Prevents bacterial growth for 24 hours
REFRIGERATION (2-8 degree celsius)
Excellent sediment preservative
Reducing agent, interieres with chemical tests for glucose, blood, leukocytes & copper reduction
Rinse specimen container with formalin to preserve cells and casts
Preservative of choice for Addis count
FORMALIN (formaldehyde)
Freserves protein & formed elements well. Does not interfere w/ routine analyses other then pH
May precipitate crystals when used in large amcunts «cloudy urine
Keeps pH about 6 0
Bacteriostatic at 18 g/L;
For culture transport, C&S
Interferes with drug & hormone analyses
BORIC ACID
Boric acid and HCl
-preserves albumin and 5-HIAA, but always choose HCl
Preserves glucose & sediments well
Interferes with acid precipitation test for protein
THYMOL
Floats on urine surface;
Clings to pipettes & testing materials
Best all-around preservative
Toluene (Toluol)
Prevents glycolysis
Good preservative for drug analysis
Inhibits reagent strip tests for glucose, blood & leukocytes
May use sodium benzoate instead of _____ for reagent strip testing
Sodium fluoride
Causes an odor change
Phenol
Convenient when refrigeration not possible
Commercial preservative tablets
Components of Saccomanno fixative
(50% ethanol + 2% carbowax)
PHYSICAL EXAMINATION OF URINE
Normal range (24 hours) ______
Average (24 hours) _______
Night urine output_______
Day: Night ratio______
Container capacity (UA)_______
Required for routine UA_____
Normal range (24 hours) 600-2000 ml
Average (24 hours) 1200-1500 ml
Night urine output <400 ml
Day: Night ratio 2-3:1
Container capacity (UA) 50 ml
Required for routine UA 10-15 ml (ave.12 ml)-urinometry and reagent strips
Increased urine volume
POLYURIA
> 2.000 mL/24 hrs (in adults - Henry
2.5 L/day (in adults - Strasinger)
2.5-3.0 mL/kg/day (in children)
Causes:
Increased fluid intake
Diuretics, nervousness
DM (sweet) ⬆️ SG (glucose)
DI (tasteless) ⬇️ SG (diluted urine)
Decreased urine volume
OLIGURIA
<500 mL/24 hrs (in adults - Henry)
<400 mL/day (in adults - Strasinger)
<1 mL/kg/hr (in infants)
<0.5 mL/kg/hr (in children)
Causes:
Dehydration
Renal diseases
Renal calculi or tumor
Complete cessation of urine flow
ANURIA
<100 mL/ 24 hrs (Graff)
Causes:
Complete obstruction (stones, tumors)
Toxic agents
Decreased renal blood flow
Excretion of more than 500 mL of urine at night
NOCTURIA
> 500 ml/night
S.G. < 1.018
Causes:
Pregnancy
Renal diseases, bladder stones
Prostate enlargement
Any increase in urine excretion
DIURESIS
Causes:
Excessive water intake (polydipsia)
Diuretics therapy, hormonal imbalance
Renal dysfunction, drug ingestion
Rough indicator of the degree of hydration and should correlate with urine S.G.
URINE COLOR
Normal urine color
Colorless to deep yellow
⬆️fluid intake=Pale yellow=⬇️SG
⬇️fluid intake=Dark yellow⬆️SG
Abnormal urine color
Red/red-brown (most common)
URINE COLOR DETERMINATION
Look down through the container against a white background (also works for determining urine clarity but not the best way)
Major pigment in urine
Lipid-soluble pigment that is a product of endogenous metabolism
Production is directly proportional to metabolic rate
UROCHROME
⬆️ in thyrotoxicosis, fever, starvation, fasting
Pink (or red)
Derived from melanin metabolism
May denosit in amornhous urates and uric acid crystals
UROERYTHRIN
(Dark yellow/orange-brown)
Derived from oxidation of colorless urobilinogen
Present in old specimens.
UROBILIN
Recent fluid consumption, polyuria, dilute random specimen
Colorless to pale yellow
Pyuria/leukocyturia (⬆️ WBCs)
Milky white
Methemoglobin (acidic urine)
Hemogentisic acid (alkaline urine): Alkaptonuria
Melanin (upon air exposure): Melanuria
Phenol derivative, Argyrol, Methyldopa/ Levodopa, Metronidazole (Flagyl)
Brown/Black
Porphyrins
Burgundy/ Purplish red/ PORTWINE
(If there’s no portwine in the choices, choose RED)
Concentrated specimen: strenuous exercise, dehydration, fever, burns.
First morning specimen
Excessive urobilin, bilirubin, carotene
Dark yellow to amber
Bilirubin has been oxidized to biliverdin
Yellow-green
Yellow-brown
Pseudomonas infection
Green
Phenol
Indican
Blue-green
Pink, Red urine
RBCs (Cloudy/ smoky red): Hematuria blood in urine
Hemoglobin (Clear red) pigments
Myoglobin (Clear red/reddish-brown/cola-colored/tea-colored)
Porphobilin (derived from porphobilinogen)
Beets (alkaline urine in genetically susceptible persons), menstrual contamination
Fuchsin (aniline dye from foods and candy)
Rifampin - most body fluids are red and TB medication
Drug that causes green color of urine
Phenol poisoning
Bright orange-red
Bright yellow
Rifampin
Riboflavin (multivitamins)
URINE CLARITY DETERMINATION
Thoroughly mix the specimen
Examine the specimen while holding in front of a light source
View through a newspaper print - BEST WAY
No visible particulates, transparent
Clear
May PRECIPITATE or be CLOTTED
Milky
Many particulates, print BLURRED through urine
CLOUDY
PRINT CANNOT be seen through urine
TURBID
Few particulates, print EASILY seen through urine
HAZY
Pathologic causes of urine turbidity
- RBCs, WBGs
- Bacteria (uniform turbidity not cleared by acidification or filtration)
- Yeasts (1DM)
- Non squamous epithelial celis
- Abnormal crystals, lymph fluid (chyluria), lipids
Non-pathologic cause of urine turbidity
- Squamous epithelial cells (⬆️ in females).
- Amorphous urates (pink sediment)
- Amorphous phosphates & carbonates
(white or beige precipitate) - Vaginal cream, semen, fecal contamination, radiographic contrast media, talcum power
Present in ACIDIC urine
Amorphous urates
Radiographic contrast media
Present in ALKALINE urine
Amorphous phosphates, carbonates
Soluble with Heat
Amorphous urates, uric acid crystals
Soluble in Dilute acetic acid
RBCs, amorphous phosphates, carbonates
Insoluble in Dilute Acetic Acid
WGCs, bacteria, yeast, spermatozoa
Soluble in Ether
Lipids, lymphatic fluid, chyle
Normal ODOR of urine due to presence of volatile acids from food
AROMATIC
FAINTLY
DISTINCT
FRAGRANT
Odorless urine
ACUTE TUBULAR NECROSIS (acute renal failure)
UTI, old urine odor
FOUL
AMMONIACAL
PUNGENT
Ketones (DM, starvation, vomiting)
FRUITY
Sweet
Maple syrup urine disease (MSUD)
CARAMELIZED SUGAR
CURRY
MAPLE SYRUP
MOUSY, MUSTY, BARNY
Phenylketonuria (PKU)
Rancid butter
Tyrosinemia
Sweaty feet, acrid
Isovaleric acidemia, glutaric acidemia
Menthol-like
Phenol containing medications
Cabbage, hops
Methionine malabsorption (Oasthouse syndrome)
Cystine disorder
Sulfur odor
Trimethylaminuria
Rotting Fish (GG)
Ingestion of onions, garlic, and ASPARAGUS, UTI
PUNGENT
Fetid
Swimming pool
Hawkinsinuria
3-hydroxy-3-methylglutaric aciduria
Cat urine
Multiple carboxylase deficiency
Tomcat urine
GLUCOSE
Principle:
Reading time:
Positive color:
Principle: Double Sequential enzyme reaction
Reading time: 30 seconds
Positive color: Green to brown
_________
Principle: Diazo reaction
Reading time:
Positive color:
BILIRUBIN
Reading time: 30 seconds
Positive color: Pink to violet
_________
Principle:
Reading time: 40 seconds
Positive color:
KETONES
Principle: Sodium Nitroprusside reaction
Positive color: PURPLE
_________
Principle: Greiss Reaction
Reading time:
Positive color:
NITRITE
Reading time: 60 seconds
Positive color: UNIFORM PINK
PROTEIN
Principle:
Reading time:
Positive color:
Principle: Protein error of indicators
Reading time: 60 seconds
Positive color: Blue-green
pH
Principle:
Reading time:
Positive color:
Principle: Double indicator system
Reading time: 60 seconds
Positive color: orange (pH 5.0) to blue (pH 9.0)
BLOOD
Principle:
Reading time:
Positive color:
Principle: Pseudoperoxidase activity of hemoglobin
Reading time: 60 seconds
Positive color:
- Uniform green/blue (Hgb/Mb)
- Speckled /spotted (intact RBCS)
UROBILINOGEN
Principle: Ehrlich reaction
Reading time: 60 seconds
Positive color: Red
_________
Principle:
Reading time: 120 seconds
Positive color:
LEUKOCYTE
Principle: LEUKOCYTE ESTERASE
Positive color: PURPLE
Principle:
Reading time: 45 seconds
Positive color:
SG
Principle: pKa change of polyelectrolyte
Positive color: Blue (1.000) to yellow (1.030)
Specimens must be returned to room temp before chemical testing by reagent strips, why?
Enzyme reactions on the strips perform best at room temp
- Dip the reagent strip briefly (____________) into a well-mixed uncentrifuged urine specimen at RT.
- Remove excess urine by _____________ to the container as the strip is withdrawn.
- _________________
- Wait the specified amount of time for the reaction to occur.
- Compare the color reaction of the strip pads to the manufacturer’s color chart in good lighting.
- No longer than 1 second
- Touching the edge of the strip
- Blot the edge of the strin on a disposable absorbent pad.
Care of reagent strips
COOL, DRY AREA
a. Store with dessicant in an opaque, tightly closed container.
b. Store below 30C (RT), do not freeze
Automated reagent strips reader principle
REFLECTANCE PHOTOMETRY
Automated reagent strips reader principle
REFLECTANCE PHOTOMETRY
the darker the color of the reagent pad, the lesser the light reflection (inversely related)
A measure of the amount of dissolved substances in a solution
Density of solution compared with density of similar volume of distilied water at a similar temp.
Influenced by number and size of particles in a solution
SPECIFIC GRAVITY
SG of random urine
1.003-1.035
1st morning urine SG
≤ 1.020
24-hour urine SG
1.016-1.022
If SG is <1.003
If SG is >1.040
Not a urine (except in D.I) 1.001
Radiographic dye present-sharp image
SG = 1.010
SG < 1.010
SG > 1.010
ISOTHENURIA
HYPOSTHENURIA
HYPERSTHENURIA
URINOMETRY (urinometer/hydrometer)
-calibration temp _____
-requires temp correction:
______ for every 3 °C that the specimen temp is BELOW the calib temp
______ for every 3 °C that the specimen temp is ABOVE the calib temp.
-requires correction for GLUCOSE and PROTEIN
1 g/dL Glucose= _______
1 g/dL Protein= ________
Urine Volume required =_________
Calibration:
[Potassium sulfate solution]
[SG reading should be _____ ]
Disadvantage:
calibration temp: 20 °C
-0. 001 for every 3 °C that the specimen temp is BELOW the calib temp
+0.001 for every 3 °C that the specimen temp is ABOVE the calib temp.
1 g/dL Glucose= -0.004
1 g/dL Protein= -0.003
Urine Volume required = 10-15 ml
SG reading should be 1.015
Disadvantage: uses large urine volume
When using the urinometer, an adequate amount of urine is poured into a proper-size container and the urinometer is added with a ______ motion.
The scale reading is then taken at the ______ of the urine meniscus.
SPINNING
BOTTOM
Refactometer is also known as?
RF
TS METER (total solids)
Put 1-2 drops of sample on the prism
Refractometry is an INDIRECT method based on _________.
Compensated to temperature________
Advantage________
Refractive index
RI= light velocity in air/light velocity in solution
Compensated to temperature (15-38 °C) or 60 °F-100 °F
Advantage: NO NEED FOR TEMPERATURE CORRECTION
REQUIRES CORRECTION FOR GLUCOSE AND PROTEIN
Calibration Reading
1. DISTILLED/deionized H2O=___________
2. 3% NaCl= ___________
4. 5% NaCl= ___________
5. 7% NaCl=___________
5. 9% Sucrose=_________________________
- 1.000 ± 0.001
- 1.015 ± 0.001
- 1.022 ± 0.001
- 1.035 ± 0.001
- 1.034 ± 0.001
Both refractometer and urinometer requires correction for ____________.
Refractometer reading is ______ than that of the urinometer by 0.002
GLUCOSE and PROTEIN
LOWER (Rf<U by 0.002)
SG DILUTION
- Specimens with very high S.G. readings can be diluted and retested
- To obtain the actual S.G., multiply the _____ of S.G. by the ______.
Decimal portion x dilution factor
Example: Urine specimen diluted 1:4 has a reading of 1.014. What is the actual S.G. reading?
Actual SG= 0.014 x 4= 0.056 (1. 056)
REAGENT STRIP for SG
REAGENT:
False +:
False -:
Add _____ to reading when pH ≥ 6.5 due to interference with brothymol blue indicator
Not affected by glucose, protein & radiographic dye (Henry)
REAGENT:
- Poly (methyl vinyl ether /maleic anhydride) BROMTHYMOL BLUE
- Ethylene glycol diaminoethyl ether tetraacetic acid BROMTHYMOL BLUE
False +: High concentration of protein (Strasinger)
False -: Highly alkaline urines (>6.5)
0.005
Obsolete method
Based on frequency of soundwave entering a soln. changes in proportion to the density of soln.
Ex: Yellow IRIS (International Remote Imaging System)
HARMONIC OSCILLATION DENSITOMETRY (H.O.D.)
IRIS DIAGNOSTICS
____ required urine volume
____ for IRIS Slideless microscope
____ for IRIS Mass Gravity Meter
Models 300 and 500 workstations
6 mL = required urine volume
4 mL (of 6 mL) = for IRIS Slideless microscope
2 mL (of 6 mL) = for IRIS Mass Gravity Meter (for S.G. determination - by using H.O.D.)
______ refers to the “sourness” of a solution, whereas _______ refers to its “bitterness”
ACIDITY
ALKALINITY
Important in the identification of crystals and determination of unsatisfactory specimens
pH
A blood pH <5.8 or >7.8 will result in death
Normal urine RANDOM pH_____
First morning pH_____
When pH is ≥ 9.0 ______
4.5-8.0
5.0-6.0
UNPRESEVED URINE
Causes of ACIDIC urine
- Diabetes Mellitus (⬆️ ketone bodies)
- Starvation (⬆️ ketone bodies)
- High protein diet
- Cranberry juice - treatment for UTI (antibaderial)
- Emphysema, dehydration, diarrhea, acid-producing bacteria (E. coli), medications
Causes of Alkaline Urine
- Renal tubular acidosis
- Vegetarian diet
- After meal - due to alkaline tide
- Vomiting - metabolic alkalosis
- Old specimens, hyperventilation, presence of wrease-producing bacteria
_______occurs after meals due to withdrawal of H ions for the purpose of secretion of H ions, Cranberry juice contains quinic acid that causes urinary excretion of hippuric acid (antibacterial).
ALKALINE TIDE
pH
REAGENTS:
Interferences:
Correlation with other tests:
Reagents: METHYL RED & BROMTHYMOL BLUE
Interferences: Runover from adjacent pads, old specimens
Correlation with other tests:
NITRITE
LEUKOCYTE
MICROSCOPIC
Most indicative of renal disease
Produces ______ in urine when shaken
PROTEIN
WHITE FOAM
NORMAL URINARY PROTEIN____
Mild/minimal proteinuria ____
Moderate proteinuria_____
Large/heavy proteinuria____
NORMAL URINARY PROTEIN
<10 mg/dL or <100 mg/ day (Strasinger), <150 mg/ day (Henry)
Mild/minimal proteinuria
<1 g/day
Moderate proteinuria
1-3 or 4 g/day
Large/heavy proteinuria
>3 or 4 g/day
Major seruin protein found in the urine
______ of plasma albumin enters the uitrafiltrate
______ of all filtered protein is reabsorbed
ALBUMIN
<0.1%
95-99%
PROTEINS in normal urine consist of
_____ ALBUMIN and _____ globulins.
1/3 albumin [95-98% reabsorb, <0.1% ultrafiltrate]
2/3 globulins
Caused by conditions that affect the plasma PRIOR to its reaching the kidney
Will not detected in routine UA
NOT indicative of actual renal disease
PRE-RENAL (BEFORE) or OVERFLOW PROTEINURIA
a. Intravascular hemolysis = ____
b. Muscle injury = _____
c. Severe infection & inflammation = ____
a. Hemoglobin
b. Myoglobin
c. ⬆️ APR’s
Proliferation of Ig-producing plasma cells
MULTIPLE MYELOMA
Immunoglobulin light chains (identical: K - K, γ-γ)
Tests = ______
Urine = precipitates at ______ (cloudy) & dissolves at _____ (clear)
BENCE- JONES PROTEIN
Tests: Serum electrophoresis, immunofixation electrophoresis
40-60 °C; 100 °C
(‘true renal disease”)
RENAL PROTEINURIA
_____________
Decreased glomerular filtration
May lead to renal failure
A. Glomerular Proteinuria
1. Diabetic nephropathy
Indicator of Diabetic Nephropathy?
MICROALBUMINURIA
- proteinuria undetectable by routine reagent step
Albumin Excretion Rate (AER) = in ug/ min or in mg/24 hours
Normal AER = ______
Microalbuminuria =_______
Clinical albuminuria=_________
Normal AER = 0-20 ug/min
NEGATIVE routine rgt. strip
Microalbuminuria = 20-200 ug/min (30-300 mg/24 hrs)
NEGATIVE routine rgt. strip
Clinical albuminuria= >200 ug/min
POSITIVE routine rgt. strip
Test for microalbuminuria
A strip employing antibody-enzyme conjugate that binds albumin
MICRAL TEST
Reagents: Gold-labeled antibody, -galactosidase, Chlorophenol red galactoside
Sensitivity: 0 - 10 mg/mL
Interference: False (-) = Dilute urine
PEINCIPLE of MICRAL test
ENZYME IMMUNOASSAY
IMMUNODIP TEST FOR MICROALBUMINURIA
Principle:
Sensitivity:
Reagents:
Interferences:
Principle: Immunochromographics
Sensitivity: 1.2-8.0 mg/dL
Reagents: Antibody coated blue latex particles
Interferences: False (-)= Dilute urine
ALBUMIN: CREATININE RATIO - CLINITEST MICROAL BUMIN STRIPS/MULTISTIX-PRO
Principle:
Reagents:
Sensitivity:
Interference:
Principle: Sensitive albumin tests related to creatinine conc, to correct for patient hydration.
Reagents
• Albumin: diodo-dihydroxydinitrophenyl tetrabromosulfonphthaleir.
• Creatinine: copper sulfate, tetramethylbenzidine, disopropylbenzenedihydroperoxide
Sensitivity:
Albumin = 10 - 150 mg/L
Creatinine = 10 - 300 mg/ dL 0.9 - 26.5 mmol/L)
Interferences: Visibly bloody/abnormally colored urine.| Creatinine = Cimetidine - False (+)
-Proteinuria when standing due to increased pressure to renal veins
-Increased venous pressure causes renal congestion and glomerular changes
-Monitored every 6 months and re-evaluated as necessary
Orthostatic / Cadet / Postural proteinuria
- young adults are mostly affected
Negative (-) FIRST MORNING
Positive (+) 2 hours after standing
ORTHOSTATIC PROTEINURIA
Negative (+) FIRST MORNING
Positive (+) 2 hours after standing
CLINICAL PROTEINURIA
⬆️ BP disorder occur during pregnancy
PRE-ECLAMPSIA
Other causes of glomerular proteinuria
Nephrotic syndrome
Toxic agents
Dehydration
Strenuous exercise
Hypertention
Amyloidosis
Pre-eclampsia
Originally discovered in workers exposed to cadmiun dust (a heavy metal).
Normaily filtered albumin can no longer be reabsorbed
Tubular Proteinuria
- Fanconi’s syndrome
- Toxic agents heavy metals
- Viral infections
Post- Renal Proteinuria (after) causes:
- Lower UTI/ inflammation
- Menstrual contamination
- Injury / trauma
- Vaginal secretions
- Prostatic fluid / spermatozoa
Protein rgt. Strips
Reagents:
False (+):
False (-):
Indicator is SENSITIVE to _______
Correlations:
Reagents:
TETRABROMphenol blue, citrate buffer at pH 3.0
Tetrachlorophenol TETRABROMosulfonphthalein, citrate buffer at pH3.0
False (+):
-high SG
-highly buffered alkaline urine
-pigmented specimen
False (-): Proteins other than albumin, microalbuminuria
Sensitive to ALBUMIN
CORRELATIONS
- Blood nitrite
- Leukocytes
- Microscopic
A cold precipitation test that reacts equally with all forms of protein.
Aka: Exton’s test
SULFOSALICYLIC ACID PRECIPITATION TEST
SSA reagent= _______
Exton’s reagent (3% SSA + sodiurn sulfate)
Procedure:
3 ml of 3% SSA + 3 mL centrifuged urine —10 mins incubation—> (+) Cloudiness
or
3 ml of 7% SSA + 11 mL centrifuged urine —10 mins incubation—>
(+) Cloudiness
No increase in turbidity
If viewed from top, circle is visible in test tube bottom
Grade:
Range (mg/dL) {stras}:
Range (Henry):
Grade: Neg
Range (mg/dL) {stras}: <6
Range (Henry): 5 mg/dL
Noticeable or perceptible turbidity.
If viewed from top, circle not visible in test tube bottom
Can read newsprint through mixture
Grade:
Range (mg/dL) {stras}:
Range (Henry):
Grade: Trace
Range (mg/dL) {stras}: 6-30
Range (Henry): 20 mg/dL
Distinct turbidity with no granulation Cannot read newsprint through mixture
Grade:
Range (mg/dL) {stras}:
Range (Henry):
Grade: 1+
Range (mg/dL) {stras}: 30-100
Range (Henry): 50 mg/dL
Turbidity with granulation but NO flocculation.
Grade:
Range (mg/dL) {stras}:
Range (Henry):
Grade: 2+
Range (mg/dL) {stras}: 100-200
Range (Henry): 200 mg/dL
Turbidity with granulation AND flocculation
Grade:
Range (mg/dL) {stras}:
Range (Henry):
Grade: 3+
Range (mg/dL) {stras}: 200-400
Range (Henry): 500 mg/dL
Clumps of protein
Grade:
Range (mg/dL) {stras}:
Range (Henry):
Grade: 4+
Range (mg/dL) {stras}: >400
Range (Henry): 1.0 g/dL or 1000 mg/dL
COMPARISON OF REAGENT STRIP AND SSA PROTEIN TEST RESULTS
Strip result: POSITIVE (+)
SSA result: NEGATIVE (-)
Highly buffered alkaline with no albumin present - false-positive reagent strip
Highly buffered alkaline with albumin present - false-negative SSA test
(To differentiate, acidify urine to pH ~ 5.0 and retest)
COMPARISON OF REAGENT STRIP AND SSA PROTEIN TEST RESULTS
Strip result: NEGATIVE (-)
SSA result: POSITIVE (+)
Proteins other than albumin present
False (+)
=Radiographic contrast media (delayed reaction)
=Drugs and/or drug metabolites (tolbutamide, penicillins, cephalosporins, sulfonamides)
(Examine precipitate microscopically - drugs and radiographic dye form crystalline precipitates; whereas protein precipitates are amorphous)
Large volume of urine can produce a _______ protein reaction despite significant proteinuria because the protein present is being excessively diluted…
S.G. should be considered in evaluating urine protein because a ____ protein in a dilute specimen is more significant than in a concentrated specimen.
Negative
Trace
Most frequently tested in urine
GLUCOSE
CLINICAL SIGNIFICANCE OF URINE GLUCOSE
⬆️ blood glucose
⬆️ urine glucose
Causes:_______
HYPERGLYCEMIA-associated
Causes:
1. Diabetes Mellitus
2. Cushing’s syndrome (⬆️cortisol)
3. Pheochromocytoma (⬆️catecholamines)
4. Acromegaly (⬆️ growth hormone)
5. Hyperthyroidism (⬆️T3, T4)
Normal blood glucose
⬆️ urine glucose
RENAL-ASSOCIATED
Causes:
1. Impaired tubular reabsorption of glucose
2. Fanconi syndrome
“ Defective tubular reabsorption of glucose and amino acids”.
Is it possible for an individual to have hyperglycemia without glucosuria?
Yes! It is possible for an individual to have hyperglycemia without glucosuria when the glomerular filtration rate is decreased due to certain diseases. Only limited amounts of glucose are able to
LL pass into the ultra-filtrate, and the tubules are able to reabsorb
all the glucose presented to them.
GLUCOSE rgt. Strip
Reagents:
False (+):
False (-):
Correlations:
Reagents:
Glucose oxidase, Peroxidase, Potassium iodide (blue to green to brown)
Glucose oxidase, Peroxidase, Tetrameihylbenzidine (yellow to green)
False (+): Oxidizing agents, detergents
False (-): High levels of ascorbic acid, ketones, high S.G., LOW TEMP, improperly preserved specimen
Glucose strip was the 1st “dip and read” reagent strip developed by Miles, Inc., in 1950
Sensitivity = 100 mg/dL
Other chromogers:
-Aminopropylcarbazole (yellow to orange-brown)
-o-toluidine (pink to purple)
Correlations: Ketones and protein
Nonspecific test for reducing sugars
COPPER REDUCTION TEST (CLINITEST / BENEDICT’S TEST)
Principle used in Copper reduction test for glucose
Copper reduction
CuSO4 (copper sulfate)—-> (+)Cu2O (copper oxide)
Blue—->Brick red
Reporting of Benedicts test
___= clear blue color, biue precipitate may form
___= reddish-yellow color, brick red or red precipitate
(-)
4+
___=bluish- green color
___=yellow-orange color, yellow-orange precipitate
Tr
3+
____= green color, green or yellow precipitate
____=yellow to green color, yellow precipitate
1+
2+
False positive
Reducing agents (ascorbic acid, uric acid)
Tip (same action as the test principle)
False negative
Oxidizing agents (detergents)
Tip (opposite of the test principle)
CLINITEST TABLET PROCEDURE
________+ 10 gtts H2O+ Clinitest tablet
-> Read reaction 15 secs after bubbling stops
5 gtts urine (5 drops)
Occurs when > 2 g/di sugar is present
Pass-through phenomenon:
• Blue > Green > Yellow > Brick red»_space;» Blue or Green-brown
• Due to reoxidation of cuprous oxide to cupric oxide and other cupric complexes (green)
• To prevent pass through, use 2gtts urine (use separate color chart to interpret the reaction)
The tablets contain:
CuSO4, =
Na citrate =
NaCO3 =
NaOH=
The tablets contain:
CuSO4, = main reacting agent
Na citrate - for heat production
NaCO3 = eliminates interfering 02
NaOH= for heat production
Glucose oxidase = 1+positive
Clinitest= negative
Small amount of glucose present
Glucose oxidase = 4+positive
Clinitest= negative
Possible oxidizing agent interference on reagent strip
Glucose oxidase = negative
Clinitest= positive
Non-glucose reducing substance present
Possible interfering substance for rgt. Strip (ex. Ascorbic acid)
Result from increased fat metabolism due to inability to metabolize carbohydrates
KETONES
Ketones renal threshold
70 mg/dL
Ketones are seen in:
Type 1 DM
Vomiting
Starvation
Malabsorption
Major ketone but not detected in reagent strip
Beta-hydroxybutyric acid
Parent ketone (1st ketone body formed)
Acetoacetic acid (AAA)/Diacetic acid (20%)
Acetone
2%
Rgt. Strip for ketone
Principle:
Reagents:
Interferences:
Correlations:
Principle:
Acetoacetic acid (acetone) + Na nitroprusside (&glycine)———-» (+) PURPLE
Reagents:
Na nitroprusside/nitreferricvanide, Clycine
False (+): Pthalein dyes, pigmented red urinc, levodopa, drugs with suifhydryl groups
False (-): Improperly preserved specimens
Correlations: GLUCOSE
ACETEST (Tablet)
________ urine + Acetest tablet
- -> (+) Purple color after 30 seconds
Composition = Sodium nitroprusside, Disodium phosphate, Glycine and Lactose
1 gtts urine
CLOUDY RED URINE
Sensitive early indicator of renal disease
Seen in: _______
Microscopic: ______
HEMATURIA
Seen in:
Glomerulonephritis
Renal calculi, tumors
Strenuous exercise trauma
Anticoagulant therapy
Intact RBC’s
Hemoglobinuria
Color of urine:
Seen in:
Microscopic:
Clear red urine
Seen in: INTRAVASCULAR HEMOLYSIS
Transfusion reactions
Hemolytic anemia
Severe burns
Brown recluse spider bites
No RBC’s seen
Heme portion of the hgb is TOXIC TO THE RENAL TUBULES
MYOGLOBINURIA
Must be at least _______, to show clear red (red-brown) urine.
Seen in:
Must be at least 25 mg/dl, to show clear red (red-brown) urine.
RHABDOMYOLYSIS
- Muscular trauma
- Crush syndrome
- Extensive exertion
- CHOLESTEROL-LOWERING STATIN MEDICATIONS
Heme portion of the myoglobin is toxic to the renal tubules
> 1.5 mg/dL= renal failure risk
What is more toxic to the renal tubules?
a. Heme
b. Hemoglobin
c. Myoglobin
C. Myoglobin
True or False:
LYSIS of RBCs in the urine usually shows a mixture of hemoglobinuria and hematuria.
TRUE
RED or PINK plasma
⬇️ haptoglobin levels
Blondheim’s test (ammonium sulfate test) ——-> PRECIPITATED
BLOOD (-)
Procedure:
Urine + 2.8g NH4Sulfate (80% satd.)
Allow the mixture to sit for 5 mins
Filter/Centrifuge
Test supernatant for blood with a reagent strip
HEMOGLOBIN
PALE YELLOW PLASMA
⬆️ CK and aldolase activity
NOT PRECIPITATED by Ammonium sulfate
BLOOD (+)
MYOGLOBIN
Rgt. Strip for BLOOD
Principle:
Reagents:
Interferences:
Correlations:
Principle: PSEUDOPEROXIDASE ACTIVITY OF HEMOGLOBIN
Reagents: =
Diisoprobylbenzene dehydroperoxide tetramethylBENZINE
Dimethyldihvdroperoxyhexane tetramethyl BENZIDINE
False +
Strong oxidizing agents, bacterial peroxidases, menstrual contamination.
False -
High SG, created cells, formalin, captopril, high concentrations of nitrite, ascorbic acid (>25 mg / dL), unmixed specimens
Uniform green / blue color =Hemoglobin / Myoglobin
Speckled / spotted
= Hematuria (Intact RBCs)
Chemstrip contains iodate overlay that eliminates ascorbic acid interference
Ilemoglobin level of 10 mg/ dL produces a positive protein reagent strip reaction
Correlations: Protein, Microscope
Early indication of liver disease
BILIRUBIN
- Tea-colored/amber/beer brown urine with yellow foam
Clinical significance:
Hepatitis
Cirrhosis
Biliary obstruction (gallstones, carcinoma)
Rgt. Strip for BILIRUBIN
Principle:
Reagents:
Interferences:
Correlations:
Principle: DIAZO REACTION
Bilirubin diglucuronide (CB) + Diazonium salt ————> Azodye
Reagents:
2,4-dichloroaniline diazonium salt
2,6-dichlorobenzene diazonium salt
False (+): Highly pigmented urines, phenazopyridine, indican, metab. of Lodine
False (-): Specimen exposure to light, high conc. of nitrite, ascorbic acid (>25 mg/dL)
(+) Tan or Pink to Violet (but VIOLET is the best answer)
Correlations: UROBILINOGEN
ICTOTEST (Tablet)
_________ + Ictotest tablet + 2 gtts H20
–> {+) _________ color after 60 seconds
ICTOTEST (Tablet)
10 gtts urine + Ictotest tablet + 2 gtts H20 –> {+) Blue to purple color after 60 seconds
- Confirmatory test; more sensitive than strip with less interference
Bile pigment that resuited from hemogiobin degradation
UROBILINOGEN
Normal value of UROBILINOGEN in urine?
Specimen____
<1 mg/dl or Ehrlich unit
Afternoon urine (2-4 pm)
Rgt. Strip for urobilinogen
Principle:
Reagents:
False (+):
False (-):
Correlations:
Principle: Ehrlich reaction
Urobilinogen (and Ehrlich-reactive compounds) + PDAB ——-> (+) Red
Reagents:
o-dimethylaminobenzaldehyde (PDAB or Ehrlich reagent)
4-methovbenzene-diazonium-tetrafluoroborate (specific for UBG)
False (+):
Ehrlich-reactive comp. (porphobilinogen, indican, methyldopa, procaine sulfonamides, p-aminosalicylic acid chlorpromazine), pigmented urine
False (-):
Old specimens, preservation in formalin high concentrations of nitrite
Correlations: BILIRUBIN
Differentiate urobilinogen (UBG), porphobilinogen (PBG) and other Ehrlich-reactive compounds (ERC).
Uses extraction with organic solvents:____&_____
WATSON- SCHWARTZ TEST
Chloroform and Butanol
Souble in CHLOROFORM & BUTANOL
Urobilinogen(UBG)
Insoluble in BUTANOL and CHLOROFORM
Porphobilinogen (PBG)
Soluble in BUTANOL
Insoluble in CHLOROFORM
Ehrlich reactive compounds (ERC)
(Inverse Ehrlich reaction)
Rapid screening test for porphobilinogen only. (≥ _____ mg/dL)
+ color:
HOESCH TEST
(≥2 mg/dL)
RED
Extravascular hemolytic disease
(Pre-hepatic hemolytic disease)
Blood:
Urine Bilirubin (CB):
Urine urobilinogen:
Blood: ⬆️ UB
Urine Bilirubin (CB): NEGATIVE
Urine urobilinogen: POSITIVE
Liver damage (hepatic jaundice)
Blood:
Urine Bilirubin (CB):
Urine urobilinogen:
Blood: ⬆️UB/CB
Urine Bilirubin (CB): +/-
Urine urobilinogen: POSITIVE
Bile duct obstruction
(Post-hepatic or obstructive jaundice)
Blood:
Urine Bilirubin (CB):
Urine urobilinogen:
Blood: ⬆️CB
Urine Bilirubin (CB): POSITIVE
Urine urobilinogen: -/+ (Strip= NORMAL)
Rapid screening test of UTI or bacteriuria
Specimen:
NITRITE
Nitrate converters are generally Grain-negative bacilli, such as the Enterobacteriaceae
Specimen: 4 hour collection or first morning urine (referred)
Rgt. Strip for NITRITE
Principle:
Reagents:
False (+):
False (-):
Correlations:
Principle: Greiss reaction
P-arsanilic acid (or sulfanilamide) + Nitrite ——-> Diazonium salt
Diazonium salt + Tetrahydrobenzoquinolin——->
(+) Uniform pink
Reagents:
Multistix = p-arsanilic acid, tetrahydrobenzo(h)-QUINOLIN-3-ol
Chemstrip = Sulfanilamide, hydroxytetrahydro benzoQUINOLINE
False (+): Improperly preserved specimens, highly pigmented urine
False (-): Nonreductase-containing bact, insufficient contact time bet bacteria & urinary nitrate, lack of urinary nitrate, large quantities of bacteria converting nitrite to nitrogen, antibiotics, high ascorbic acid, high SG
Pink spots/edges = considered NEGATIVE
(+) Nitrite corresponds to 100,000 organisms/ mL
If the nitrite test area shows a negative reaction, UTI cannot be ruled out
(Some UTIs are caused by Gram (+) cocci & yeasts - they lack nitrate reductase enzyme)
Dietary nitrate can be found in green vegetables
Correlations: protein, leukocytes, microscopic
Significance:
Urinary tract infection or inflammation
Screening of urine culture specimens
LEUKOCYTES
Rgt. Strip for Leukocytes
Principle:
Reagents:
False (+):
False (-):
Correlations:
Principle: Leukocyte esterase
Indoxylcarbonic acid——> Indoxyl + Acid indoxyl + Diazonium salt ——> (+) Purple
Reagents: =
Derivatives pyrrole amino acid ester, Diazonium salt
Indoxylcarbonic acid ester, Diazonium salt
False (+): Strong oxidizing agts, formalin, highly pigmented urine, nitrofurantoin
False (-): High concentrations of protein, glucose, oxalic acid, ascorbic acid
With esterase: Neutrophil, Eosinophil, Basophil, Monocyte, Histiocyte, Trichomonas
No esterase: Lymphocyte
Strip can detect lysed WBCs.
Trichomonas, Chlamydia, yeast, & interstitial nephritis produce pyuria w/o bacteriuria
Correlations: protein, nitrite, microscopic
Water-soluble vitamin
11th Reagent Pad
Brands:
C-stix=_____
STix=_____
others: VChem. Urispec GP + A, and Merckoquant
ASCORBIC ACID
Ascorbicacid (2 5 mg/dL) + Phosphomolybdate ——> (+) Molybdenum blue
Brands:
C-stix= 10 seconds
STix= 60 seconds
GC-MS= more accurate quantitative method
Ascorbic acid FALSE NEGATIVE reactions on: BB LNG
Blood
Bilirubin
Leukocyte
Nitrite
Glucose
