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ENZYME TESTS
 What is an enzyme?
 A biological catalyst
 It increases the rate of reaction by lowering the activation
energy
 They are proteins
 Enzymes can be denatured by temperatures, pH
 Enzymes bind to reactants ( substrates) . Where the enzyme
and the substrate are bind is called the (active site).
 The active site and substrate shape are complementary which
enable them to bind.

Source:
http://www.medic8.com/blood-disorders/blood-test/types/blood-enzyme-tests.html
 Graph showing activation energy

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 Enzyme Tests
Enzymes are not only used to increase the rate at which a reaction
takes place, but it also used in the medical field to diagnose various
illnesses. Some of the enzymes that are used in the medical diagnosis
are:
• Alkaline Phosphatase
• Acid Phosphatase
• Serum Amylase
• Alanine Transaminase
• Aspartate Transaminase

 The prefix of the word tell the substrate which will be worked on and the
suffix –ase is then added to give the name of the enzymes.
 Alkaline phosphatase (Alp)
 It is called Alkaline Phosphatase because of its ability to function in
alkaline conditions

 Alkaline Phosphatase is found throughout the body.

 It is mostly found in the liver, bone , placenta and the intestines. Each
produce a slight variation in alkaline phosphatase called ISOENZYME
(each of two or more enzymes with identical function but different
structure).

 In the laboratory, alkaline phosphatase is measured as the total amount

or the amount of each of the four isoenzymes. Methods to measure them
separately are based on these differences.

 The normal activity of Alkaline Phosphatase is 13-38 U/L which is found

in the serum (a component of blood)
 If the activity goes below and above the normal then the patient might
be diagnosed with Liver or Bone disease.

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 Each Isoenzyme reacts differently chemically

 Alkaline phosphatase level maybe affected by:

 the intake of fatty food
 pregnancy
 growing children
 liver problem
 menopause
 age

 Bone and liver disease increase the level of alkaline phosphatase up

to five times more
 Preparation of the
 The patient should fast for 8 to 10 hours prior to the test
patient
 Should note if the patient is pregnant

 Should also note if the patient have bone disease such as:
 Rickets
 Any history of heart failure
 Celiac disease caused by malnutrition

 Protein deficiency
It Acid phosphatase
hydrolyses the phosphate group

Itis also found throughout the body such as blood, bone

and prostate glands.

Itscalled Acid Phosphatase because of its ability to

function in a highly acidic environment.

The normal activity of acid phosphatase is 2.5-12 U/L

which is found in the serum

Abnormality of this activity (above or below the level) signal

the onset or progression of prostate cancer.
 Its concentration is 100 times higher in the prostate glands

 Acid phosphatase level may vary depending on age, medication (antibiotics)

 It is also a valuable presumptive test for the screening of swabs collected

from sexual assault survivors and as well as for the testing of stains found on
clothing and bedding.

Preparation of the patient

 The patient should not do rectal exam 2 to 3 days prior to test

 No prostate massage for 2 to 3 days prior to test

 No fasting is required


 Serum amylase
Found in the blood

 Amylase hydrolyses amylose in starch into sugar

 It is found in the pancreas and salivary glands. When the

pancreas is diseased or inflamed, amylase releases into the
blood and a test can be done to measure the level of this
enzyme in your blood. Amylase can also be measured with a
urine test.
 It is also perforated peptic ulcer, torsion of an ovarian cyst,
strangulation ileus, macroamylasemia and mumps. Amylase
may be measured in other body
fluids,including urine and peritoneal fluid.

 The normal activity is 19-80 U/L in the serum. If it goes below

or above the normal activity pancreatic disease or mumps will
Preparation of the patient
 Amylase in the blood maybe increase by:
 birth control, aspirin, morphine

 Patient should fast at least 5 hours before the test

 Will be educated why the test is recommended

Alanine

transaminase (ALT)
Called Serum Glutamic Pyruvic Transaminase (SGPT) or
Alanine aminotransferase (ALAT)

 Found in the serum of the blood

 Alanine is an amino acid

 During transamination the amino group is removed from the

amino acid where it is attached to  α-ketoglutarate where  α-
keto-acid and glutamate are formed.

 Glutamate is the new amino acid form



 Commonly linked to liver

 Small amount are found in the kidney, heart, pancreas and

muscles

 The normal activity of alanine transaminase is 3-17 U/L . If

this activity goes below or above Hepatitis disease is
diagnosed.

 This test is often done with other tests such as Aspartate

aminotransferase (AST), Alkaline Phosphatase, Lactate
Dehydrogenase(LDH)
 
Preparation of the
Avoid strenuous exercise before doing the test
patient
 Medications alter the results so the patient is required to stop
taking them such as Metronidazole and Tinidazole for several
days before having an ALT test. done.

 Allergies should be notified

 Should also note if patient is pregnant and smoke cigarettes

 Aspartate Transaminase
This enzyme disobey the rule of transamination
(AST)
 The amino group that is detached from the amino acid does not
combine with Alpha Ketoglutamate but Oxaloacetate to form
Aspartate

 Formerly Aspartate Transaminase was called Serum Glutamic

Oxaloacetic Transaminase (SGOT).

 Usually found in red blood cells, liver, heart, muscle tissues,

pancreas and kidney.
 When Hepatocellular disease( liver disease) or coronary occlusive
heart disease( where there is an obstruction in the flow of blood) is
suspected AST test is done.

 AST is also useful for the diagnosis of myocardial infarction (MI) along
with the observation of the enzyme creatine phosphokinase and
lactate dehydrogenase. 

 The normal activity of aspartate transaminase is 7-19 U/L in the

serum. While in the cerebrospinal fluid its 7-49 U/L
Preparation oftothe
 Explain the procedure patient
the patient

 Discuss with the patient the need and reason for frequent
venepuncture in diagnosing myocardial infarction (MI) ( is that
the AST level uses within 6 to 10 hours after MI, peaks at 12
to 48 hours and return to normal after 3 to 4 days) 

 Inform the patient not to do any strenuous exercise (because

it may cause increase levels  of AST)

 Also if the patient is on any medication that may alter the

results (such as cholinergic agents, hepatotoxic medication,
verapamil, methyldopa, isoniazid, opiates, erythromycin)
inform the patients not to take any 12 hours before the test.

 No fasting is required
 How is the test done?

 For all the enzymes that we have mentioned serum is needed to test
for the level of the activity of that enzymes. Serum is the liquid that
appears as a straw yellow that can be separated from the blood
without any clotting factors. Plasma differs from serum in that it
contains erythrocytes ( red blood cells) , leukocytes( white blood
cells), thrombocytes (platelets) , dissolved proteins and minerals. It is
the clotting factors that makes the difference between serum and
plasma. So a blood sample must be taken from patient to a test on the
serum.
 How to do a blood test?
2. The doctor then review the patient’s
1. The doctor would introduce document and then explain why the test is
him self to the patient and recommended even though patient would
state what will be done. have been educated about it before. If the
patient agrees he then proceeds.
 4. The doctor gathers all the supplies he
3.The doctor then washes needs; tourniquet, alcohol swab that contains
his hand with antimicrobial 70% alcohol, sterile needle, red top tube and
bandage.
soap then dry them. Then
He allow the patient to sit in a chair with an
put his gloves on. arm rest where he search for the vein while
communicating with the patient
5. A tourniquet is than tied around 6. The patient then forms a
the upper arm of the patient to fist. The patient should not
make the vein more visible. pump while doing so.
7. He then locate the area 8. He then cleans the area with
where the vein is again. the alcohol swab for 30 seconds.
 9.The sterile needle is then inserted
in the vein where 5-10mL of blood is 10. The area is cleaned again after
drawn in a red top tube. The the needle is removed. Pressure is
tourniquet is removed while the blood applied to the area. The area is
is collecting in the tube. then covered with a band aid

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 12. The tube is the labeled
11. The needle is then with the patient’s name, the
disposed of in a sharp’s box. type of test, date and time.

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 Handling of samples

 The blood samples are then stored at 30-37 degrees Celsius

which are then transported to the laboratory as quickly as possible

 The tube is allowed to stand for 30 minutes which allows the blood
to clot to separate the serum from the blood cells prior to
centrifugation.

 The serum should be separated from the blood within two hours of
collection.
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clotting.jpeg
 The blood in the tube is then placed in the centrifuge for 10
minutes where the blood components are further separated
into serum, white blood cells and red blood cells.

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 Interpretation of
 When the serum has been removed from the other blood
results
components; it is reacted with various chemical to test for the
enzymes activity.

 Once the enzymes (the ones that was discussed earlier) activity
has been detected it is compared to the normal activity (control).
However, the results may vary from one individual to next and
laboratory to laboratory.

 With alkaline phosphatase test results may varies because of age,

gender, certain medications, pregnancy, malnutrition and protein
deficiency. So all of these factors should be taken in to
consideration when diagnosing bone and liver disease.
 Acid phosphatase the age of the male should always be concerned,
health, and activities. If these are not taken into account diagnosing of
prostate cancer can be wrong.

 For serum amylase the patient should not eat several hours before the
test, the eating habit should be considered, diet, age and places
frequently visited in diagnosing pancreatic disease and mumps.

 In Alanine transaminase patient should avoid strenuous exercise,

certain medications , and allergies should be noted. So when
comparing the results to the normal those can be taken into
consideration for Hepatitis.
 Finally, for Aspartate Transaminase activities of the individual
should be considered, pregnancy, chronic renal dialysis, Beri
Beri, acute renal disease age and gender before diagnosing
heart attack and hepatitis.
 references
• Article from medlineplus.http
://www.nlm.nih.gov/medlineplus/ency/article/003473.htm
• WedMD
http://www.webmd.com/digestive-disorders/alanine-aminotransfe
rase-alt
• Cigna
http://www.cigna.com/healthwellness/hw/medical-tests/alanine-
aminotransferase-hw20645
• W.H.O Laboratory Services
http://www.who.int/hiv/pub/imai/om_8_laboratory_services.pdf
• XpressBio Life Science Products
http://www.xpressbio.com/sites/default/files/3460-01-XPRESSBIO-
Alanine%20Transaminase-ALT-Enzymatic%20Assay%20Manual.pdf