Special tests In urine

Salah Aldeen Abass Ibrahim

MSc clinical chemistry
Introduction
Urine is ultrafiltrate of plasma produced after
successful tightly controlled filtration,Reabsorption
and secretion process which take place in renal
system specifically nephrones.Contains mostly
water 💧 along with waste products (urea ,creatinine,
uric acid,Bilirubin, end products of
metabolism) ,undesired metabolites (electrolytes
Na,K,Ca,Mg, Zn,P,Fe,Ag,Mn,Cl,HCO3 ,Hydrogen
ions),glucose, abnormal proteins and scaping good
ones like amino acids and albumin.
After being secreted to collecting duct of renal
tubules urine travel to the bladder through pare of
ureters left ureter from the left kidney and right from
the right kidney,and stay in that bladder until being
released by urination which is controlled by physical,
mechanical and mostly by Neurological actions
through urethra.
So urine is a mirror reflecting what is going on in your
plasma and due to the simplicity and noninvasive
collection of urine it's considered a routine,
simple,cost-effective and highly informative
laboratory test which is performed for screening
purposes mostly and also to diagnose, monitor
and/or follow-up and treatment of patients
• The most routinely carried out investigations on urine
includes:
• Inspection of urine color since urine color varies
depending on the overall health condition and the
quantity & quality of what that urine contains.
• Inspection of urine odor since normal urine has
aromatic smell that can change into other smells
according to what is dissolved in urin.
• Clarity of the urine also important because turbid
urine means that there is undisolved particles, cells or
organisms in urine.
• Chemical tests for pH, specific gravity (how much
concentrated this urine is ) protein (mostly albumin)
ketones, Glucose Bilirubin and urobilinogen.
• Microscopic examination of urine sediment to quantify
and evaluate the particles components of the urine
• So routine test are general ,flash screening look into
urine and according to tests results and patient’s
particular illness further close-up and much
intensive work on urine can be done to reveal the
truth of excluding or supporting a suggested
diagnosis.
• And remember special tests are carried out only
when the routine one is inconclusive
Newborn Screening
Tests
performed primarily to detect and monitor newborns for (inborn
errors of Metabolism)IEMs. because many of these disorders cause
the buildup of unmetabolized toxic food ingredients, it is important
that the defects be detected early
in life. Levels of these substances are elevated more rapidly in
blood than urine.
and includes : -
Amino Acid Disorders
Porphyrin Disorder.
Mucopolysaccharide Disorders.
Purine Disorders
Carbohydrate Disorders
Amino acids disorders
• The amino acid disorders with urinary screening
tests include:
• phenylketonuria (PKU)
• :PKU is caused by failure to inherit the gene to
produce the enzyme phenylalanine hydroxylase.The
gene is inherited as an autosomal recessive trait.the
disorder lead to accumulation of phenylpyruvic acid
which causes mousy odor. chloride reaction
performed by tube test. The addition of ferric
chloride to urine containing phenylpyruvic acid
produces a permanent blue-green color.
Tyrosyluria:accumulation of tyrosine in blood due to
metabolic or inherited defects ,the urine may
contain excess tyrosine or its degradation products,
p-hydroxyphenylpyruvic acid and p-
hydroxyphenyllactic acid.detected in urine by using
nitroso-naphthol
1. Place five drops of urine in a tube.
2. Add 1 mL of 2.63N nitric acid.
3. Add one drop of 21.5% sodium nitrite.
4. Add 0.1 mL 1-nitroso-2-napthol.
5. Mix.
6. Wait 5 minutes(orange red color).
Melanuria tyrosine has other metabolic pathways
responsible for the production of melanin,
thyroxine, epinephrine, protein, and tyrosine
sulfate.Deficient production of melanin results in
albinism while Increased urinary melanin darkens
the urine. The darkening appears after the urine is
exposed to air.
Alkaptonuria urine from patients with
this condition darkened after becoming alkaline
from standing at room temperature. so it called
alkali lover,” or alkaptonuria, defect in the
phenylalanine-tyrosine pathway and occurs from
failure to inherit the gene to produce the enzyme
homogentisic acid oxidase.Paper and thin layer
chromatography procedures are available for
quantitating homogentisic acid. The silver nitrate
test for homogentisic acid (dark color appears after
adding 10% NH4OH .
Branched-Chain Amino Acid Disorders

Maple syrup urine disease (MSUD) is rare and caused by an

IEM, inherited as an autosomal recessive trait. The amino acids
involved are leucine,isoleucine, and valine. the accumulation
keto acids in the urine produces strong odor resembling maple
syrup.The 2,4-dinitrophenyl hydrazine (DNPH) urine screening
test (turbid urine)
Organic Acidemias : symptoms of the organic acidemias include
early severe illness, with vomiting& metabolic acidosis;
hypoglycemia; ketonuria; and increased serum ammonia.The
three most disorders are isovaleric, propionic, and
methylmalonic acidemia.
Isovaleric acidemia possess a characteristic odor of “sweaty
feets) which is caused by the accumulation of isovalerylglycine
due to a deficiency of isovaleryl coA in the leucine pathway.
Tryptophan Disorders

hydroxyIndoleacetatic acid 5-
HIAA and indicanuria are the
m9st common tryptophan
metabolism disorders .can tested
in urine by Adding nitrous acid
and 1-nitroso-2-naphthol to urine
that
contains 5-HIAA causes the urine
to turn purple to black,.
Cystinuria failure to reabsorb in kidney leat to excess and
detected by using nitroprusside. Reduction of cystine by
sodium cyanide followed by the addition of cyanide
nitroprusside produces a red-purple color (ketones and
homocystine can cause false positive results) .
Homocystinuria
Defects in the metabolism of the amino acid methionine
produce an increase in homocystine throughout the
body.following a positive cyanide-nitroprusside test
confirmatory silver nitroprusside should be done thus only
homocystine is reacted.
Porphyrin Disorder
Porphyrins are the intermediate compounds in the
production of heme.Disorders of porphyrin metabolism are
collectively termed porphyrias. They can be inherited or
acquired from RBCs and hepatic malfunctions or exposure to
toxic agents.
. Red color urine can be seen in porphyrias The two screening
tests for porphyrinuria use the Ehrlich reaction and
fluorescence under ultraviolet light in the 550- to 600-nm
range. The Ehrlich reaction can be used only for the detection
of ALA and porphobilinogen. Acetyl acetone must be added
to the specimen to convert the ALA to porphobilinogen prior
to performing the Ehrlich test. The fluorescent technique
must be used for the other porphyrins.
Urine for reducing
substances
Positive copper reduction test result combined with a
negative reagent strip glucose oxidase test result is
strongly suggestive of a disorder of carbohydrate
metabolism (melituria including
glactosuria,lactosuria and fructose).
Urine for quantitative
electrolytes analysis
Electrolytes as sodium, potassium, calcium,
magnesium, phosphate and others can be measured
in urine samples to rule out or support particular
illness of renal or other organs origin.
And for flowup of treatment and monitoring for
critical care patients with kidney disease or heart
failure,and acid-base disorders.
Urine for drugs

• * Purpose: To detect the presence of drugs or their

metabolites in urine.
• * Methods: Immunoassays and gas chromatography-
mass spectrometry (GC-MS).
• * Drugs detected: Opiates, amphetamines, cocaine,
marijuana, PCP, barbiturates, benzodiazepines, and more.
• * Factors affecting detection: Drug type, frequency of
use, individual metabolism.
• * Uses: Pre-employment screening, post-accident
testing, court-ordered testing, medical monitoring.
Urine for osmolality
test that measures the concentration of particles in
your urine. These particles include electrolytes, urea,
and other substances. It's a valuable tool for
evaluating kidney function and diagnosing electrolyte
imbalances, and diabetes insipidus.
Urine osmolality is typically measured in milliosmoles
per kilogram of water (mOsm/kg). A higher
osmolality indicates a more concentrated urine,
while a lower osmolality indicates a more dilute
urine.
Urea and Creatinine in
urine
• Clinical Significance of Urea and Creatinine in Urine
• * Urea: High levels can indicate kidney failure or
liver disease.
• * Creatinine: Elevated levels suggest impaired
kidney function.
• * U/C ratio: Helps differentiate between prerenal
and postrenal causes of acute kidney injury.
Urine for pregnancy
testing
• * Principle: Detect hCG in urine.
• * Techniques:
• * Immunochromatography: Most common, uses antibodies to
detect hCG.
• * Enzyme-linked immunosorbent assay (ELISA): More
sensitive,.
• * Types: Home tests (OTC) and professional tests.
• * Timing: Best taken first thing in the morning.
• * Accuracy: Can be influenced by factors like timing and test
quality.
• * Confirmation: Positive results should be confirmed by
ultrasound.
Urine for cytology
screening
• * Purpose: To detect abnormal cells in urine, primarily used
for bladder cancer screening.
• * Technique: Cells are collected, stained, and examined under
a microscope.
• * Indications: Hematuria, urinary symptoms, risk factors for
bladder cancer, such as( smoking, exposure to certain
chemicals, or a family history of the disease), follow-up.
• * Detections: Bladder cancer, infections, inflammation,
benign conditions.
• * Limitations: False negatives, false positives, may require
further testing.
Urine for culture and
sensitivity
* Purpose: To identify bacteria or fungi in a urine sample and
determine their susceptibility to antibiotics.
* Common Uses: Diagnosis and treatment of urinary tract
infections (UTIs).
* Sample Collection: Clean-catch midstream, catheterization,
or suprapubic aspiration.
* Laboratory Techniques: Inoculation in Blood
agar ,CLED ,MacConkey agar, incubation at (37°C) for 24-48
hours., colony counting, identification, and sensitivity testing.
* Results: Positive (bacteria or fungi identified) or negative (no
microorganisms found).