Group 4

URINE SCREENING FOR

METABOLIC DISORDERS
Two Category That Causes The
Appearance Of Abnormal Substances In
The Urine

1. Overflow Type
• Results from disruption of normal metabolic
pathway
2. Renal Type
• Caused by malfunctions in the tubular
reabsorption mechanism
Associated with urinary
overflow
1. Fats metabolism
2. Protein metabolism
3. Carbohydrates metabolism
4. Inborn error metabolism
 Failure to inherent the gene to produce enzyme which leads
to enzyme disruption
5. Organ Malfunction
Abnormal Metabolic Constituents Or Conditions
Detected In The Routine Urinalysis
COLOR ODOR CRYSTALS
Homogentisic Phenylketonuria Cystine
acid
Melanin Maple Syrup Leucine
Urine Disease
Indican Isovaleric Tyrosine
Acidemia
Porphyrins Cystinuria Lesch-Nyan
Cystinosis disease
Homocytinuria
 Major Disorders Of Protein And Carbohydrate Metabolism Associated With
Abnormal Urinary Constituents As To Functional Defect
OVERFLOW
METABOLIC RENAL
INHERITED
Phenylketonuria Infantile tyrosinemia Hartnup disease
Tyronisemia Melanuria Cystinuria
Alkaptonuria Indicanuria
5-Hydroxyindoleacetic
Maple syrup disease acid
Organic acidemias Porphyria
Cystinosis
Porphyria
Mucopolysaccharidoses
Galactosemia
Lesch-Nyan disease
Newborn screening tests
1. Performed primarily to detect and monitor
newborns for IEMs
2. Almost 29 metabolic disorders are required
to be tested
3. Disorders must be detected early in life
because it can cause build up of
unmetabolized toxic food ingredients
Six conditions tested

1. Congenital Hyperthyroidism
2. Congenital Adrenal Hyperplasia
3. Galactosemia
4. Phenylketonuria
5. Maple Syrup Urine Disease
6. Glucose-6-phosphate dehydrogenase
deficiency
Screening test

SPECIMEN COLLECTION
 Blood collection on infants heel
TESTING
 MS/MS-Tandem Mass Spectrophotometry
• Capable of screening the infant blood sample
for specific substance associated with particula
IEMs
Phenylalanine-tyrosine disorders
PKU
Tyrosyluria
Alkaptonuria
Melanuria
Phenylalanine-tyrosine disorders

1. Phenylketonuria
 Most common and well known
aminoaciduria.
 Caused by the failure to inherit gene
coding the enzyme Phenylalanine
hydroxylase.
 Urine is mousy odor.
 Results in severe mental retardation if
left undetected.
Baby with PKU
Screening test for PKU

Guthrie’s test
 Blood from heel stick placed
on filter paper disk
 The disk was placed in the
culture of Bacillus subtilis
incorporated with inhibitor
beta-2 thienylalanine.
 Phenylalanine counters the
action of the inhibitor, thus
promoting the bacterial
growth.
 Positive result: Bacterial
growth around the disk.
test for PKU

•Ferric chloride Tube Test

 1 mL of urine of placed
in a tube
 Add five drops of 10%
FeCl3
 Positive result: Blue-
green color
Phenylalanine-tyrosine disorders

2. Tyrosyluria
 Result from the
urinary overflow of
Tyrosine
(Tyrosinemia) in the
plasma.
 Acquired in both
liver disease and
hereditary
disorders.
 Rancid Butter – odor
of urine
Types of tyrosinemia

1. Type I
 Caused by the deficiency
of the enzyme
Fumarylacetocetate
hydrolase (FAH).
 Type I produces a
generalized renal tubular
disorder and progressive
liver failure in infants
soon after birth.
Types of tyrosinemia

2. Type II
 Lack of enzyme
Tyrosine
aminotransferase.
 Development of
corneal erosion and
lesion on the palms.
Types of tyrosinemia

3. Type III
 Lack of enzyme p-
hydroxyphenylpyruvic
acid dioxygenase.
 This may lead to
mental retardation.
test for tyrosyluria

1. Nitroso-napthol Test
 (+) Orange-red
2. 2-4 DNPH
 (+) Yellow
3. 10% FeCl3
 (+) Transient Green
Phenylalanine-tyrosine disorders

3. Alkaptonuria
 Caused by the
failure to inherit
gene to produce
Homogentisic acid
oxidase.
 Urine darkens after
becoming alkaline.
 Brown pigments
deposit can lead to
arthritis and liver
disease.
Test for alkaptonuria

1. 10% FeCl3 tube test

 (+) Transient Blue
2. Benedict’s/ Clinitest
 (+) Yellow precipitate
3. Homogentisic acid test
 (+)
Phenylalanine-tyrosine disorders

4. Melanuria
 Result from the
urinary overflow of
Tyrosine
(Tyrosinemia) in the
plasma.
 Acquired in both
liver disease and
hereditary
disorders.
 Rancid Butter – odor
of urine
Branched chain amino acid
disorders
MSUD
Organic Acidemias
BRANCHED CHAIN AMINO ACID
DISORDERS
Differ from other
amino acids by having a
methyl group that
branches from the main
aliphatic carbon chain.
Significant laboratory
finding is the presence of
ketonuria in a newborn.
BRANCHED CHAIN AMINO ACID
DISORDERS
1. Maple Syrup Urine Disorder
 Inherited as autosomal recessive trait
 Involves Leucine, Isoleucine, and Valine
 Failure in transamination of these3 amino
acids.
 Deposition of keto acids in the blood and
urine.
 Maple syrup odor of urine.
 Failure to thrive after approximately1 week.
Test for msud

1. 2,4-Dinitrophenylhydrazine
(DNPH) Reaction
 Not specific for MSUD
(PKU has high keto acids
too).
 Large doses of Ampicillin
interfere.
 Positive strip result =
Positive DNPH
 (+) Yellow precipitate
BRANCHED CHAIN AMINO ACID
DISORDERS
2. Organic Acidemia
 Characterized by:
 Severe illness
 Vomiting
 Metabolic acidosis
 Hypoglycemia
 Ketonuria
 Serum ammonia
 ISOVALERIC PROPIONIC METHYLMALONIC
ACIDEMIA ACIDEMIA ACIDEMIA
Isovaleryl
Errors in the metabolic pathway
coenzyme A
converting isoleucine, valine,
CAUSE deficiency in the
threonine, and methionine to
leucine pathway.
succinyl coenzyme-A.

sweaty-foot
URINE
urine odor
TEST MS/MS MS/MS MS/MS
P-nitroaniline + Urine (methylmalmonic acid)
RXN
 emerald green
Three Disorders Under Acidemias
Tryptophan disorders
Indicanuria
5-HIAA
Tryptophan disorders

1. Indicanuria
 Certain intestinal disorders may increase the
amount of tryptophan that are converted into
indole.
 Excess indole reabsorbed and converted to
indican and then excreted in the urine
 Also known as Blue Diaper Syndrome, when
indican is excreted in the urine it is colorless
until oxidized to the dye indigo blue by
exposure to air
Tryptophan disorders

2. Hartnup Disease
 Inherited defect that
affects not only the
intestinal reabsorption
of tryptophan but also
the renal tubular
reabsorption of other
amino acids resulting
in amino aciduria.
Tryptophan disorders

3. 5-Hydroxyindoleacetic acid
 Normally the body uses most of the serotonin
and only small amounts of its degradation
product 5-HIAA are available for excretion in the
urine.
 Carcinoid tumors involving argentaffin cells
develop, excess amounts of serotonin are
produced resulting in elevation of urinary 5-HIAA
levels.
TEST FOR 5-HIAA
1. Silver nitropprusside test
 Random or First morning specimen
 (+)
 Normal daily excretion: 2-8mg
 Excretion>25mg/day : argentaffin cell tumors
Cystine disorders
Cystinuria
Cytinosis
Homocystinuria
Cystine disorders

1. Cystinuria
 Marked by elevated amounts of the amino acid
cystine in the urine.
 Increased urinary cystine is caused by the
inability of the renal tubules to reabsorb cystine
filtered by the glomerulus.
 Modes of inheritance: reabsorption of cystine,
lysine, arginine and tyrosine is affected.
 Reabsorption of cystine and lysine is affected.
TEST FOR CYSTINURIA

1. Cyanide nitroprusside
 (+) red purple color
2. Microscopic examination
 Observation of cystine crystal in first morning
urine.
Cystine disorders

2. Cystinosis
 Prevents the release of cystine into the cellular
cytoplasm for metabolism.
 It affects many areas of the body including the
cornea, bone marrow, lymph nodes and internal
organs.
 Major defect in the renal tubular reabsorption
mechanism also occurs.
 CYSTINE DISORDERS
•Infantile Nephropathic Cystinosis
 Rapid progression of renal failure
 Late on set Nephropathic Cystinosis
 Gradual progression to renal failure
 Nonnephropathic Cystinosis
 Relatively benign but may cause some ocular
disorders
Cystine disorders

3. Homocystinuria
 Defects in the metabolism of methionine produce
an increase in homocystine throughout the body.
 Increased homocystine can result in failure to
thrive, cataracts, mental retardation,
thromboembolic problems and death.
 Cyanide nitroprusside (+) and Silver
nitroprusside(+) = homocystinuria.