AMYLASE LIPASE LACTATE DEHYDROGENASE (LDH) TROPONIN I

(DiaSys) (DiaSys) (DiaSys) (Finecare)

Wavelength 410 nm 571 nm 340 nm
Measurement Kinetic Kinetic Linear kinetics
 Serum  Whole blood
 Serum  Seru,
Sample  Heparin plasma  Serum
 Heparin plasma  Heparin plasma
 Urine  Plasma
Function: Function: Cardiac Troponin I (cTn I) is a
Lipases are enzymes which Lactate dehydrogenase (LDH) is cardiac muscle protein with a
Function:
hydrolyze glycerol esters of an enzyme, consisting of five molecular weight of 22.5
a-Amylases are hydrolytic
long fatty acids. different isoenzymes, which kilodaltons.
enzymes which break down
catalyze the interconversion of
starch into maltose
 The enzyme and its cofactor L-lactate and pyruvate.  Troponin complex:
colipase are produced in the - Troponin T (Tn T)
Origin:
pancreas.  Present in the cytoplasm of all - Troponin C (Tn C)
1. Pancreas
 Lipase is secreted in small human tissues. - Troponin I (Tn I)
Summary (Pancreatic amylase)
amounts by: Higher concentrations in: > Play a fundamental role in the
> released to intestinal tract
1. Salivary glands 1. Liver transmission of intracellular
2. Salivary glands
2. Gastric, pulmonary, and 2. Heart calcium signal actin-myosin
(Salivary amylase)
intestinal mucosa 3. Skeletal muscle interaction in the heart.
> secreted to saliva
Lower values in:  The human cTn I has an
 Bile acids and colipase --> 1. Erythrocytes additional amino acid residues
Elimination/Excretion:
form micellar complexes with 2. Pancreas on its N-terminal that do not
Kidney --> urine
the lipids and binds lipase on 3. Kidney exist on the skeletal forms thus
the substrate/water interface. 4. Stomach making it a specific marker to
 indicate cardiac infarction.
 Elevation of amylase activity in
serum = rise of urinary
amylase activity *in relation to Acute Myocardial
 For diagnosis of pancreatic  For investigation of pancreatic Infarction
disorders disorders cTn I CK-MB
 Increased LDH
 Detecting the development of  Acute pancreatitis - 2-50 fold After 3-6 hrs 4-6 hrs
1. Myocardial infarction
complications increase in the upper onset
2. Cancer
 Acute pancreatitis - reference limit within 4-8 Returns 5-7 days 36-48 hrs
3. Diseases of liver, blood, or
increased blood amylase hours after the beginning of to normal
muscle
activity within few hours after abdominal pain. Remains
Clinical
onset of abdominal pain. Peak: 24 hours elevated
Significance  Not organ-specific
Peak: appx. 12 hours Decrease: 8-14 days for 6-10
 Determination of its
Returns within reference range: days
isoenzymes or other
After 5 days  Other diseases with increased Peak 14-20 hrs
enzymes (ALP, ALT, AST) is
 Non-pancreatic diseases with lipase values:
necessary for differential
high amylase levels: 1. Chronic pancreatitis  Level of cTn I is very low in
diagnosis.
1. Parotitis 2. Obstruction of pancreatic normal healthy people, and
2. Renal insufficiency duct not detected in patients with
 Confirmation of acute skeletal muscle injury.
pancreatitis through lipase
measurement.
Method Enzymatic photometric test Enzymatic color test Optimized UV-test Fluorescense immunoassay
The substrate 4,6-ethylidene-(G7)- A synthethically produced lipase The FineCare cTn I Rapid
Principle
p-nitrophenyl-(G1)-a-D- substrate (1,2-o-dilauryl- Quantitative Test uses a sandwich
maltoheptaoside (EPS-G7) is
cleaved by a-Amylases into
various fragments.
These are further hydrolyzed in a
second step by a-Glucosidase
producing glucose and p-
nitrophenol. The increase in
absorbance represents the total
(pancreatic and salivary) amylase
activity activity in the sample.
racglycero-3-glutaric acid-(6-
methylresorufin) ester) is added to
a micro-emulsion which is
specifically split by lipase in the
presence of colipase and bile
acids. The combination of lipase
and bile acids make this specific
and reliable for pancreatic lipase
without any reaction due to lipolytic
enzymes or esterases. The
reagent composition has been
thoroughly optimized to avoid
serum matrix effects. The
generated methylresorufin ester is
spontaneously degraded to
methylresorufin. The absorbance
by this red dye is directly
proportional to the lipase activity in
the sample.
immunodetection method. When
sample is added into the sample
well of the Test cartridge, the
fluorescence-labeled detector anti-
cTn I antibodies on the sample
pad bind to cTn I antigens in blood
specimen and they form immune
complexes. As the complexes
migrate on the nitrocellulose matrix
of test strip by capillary action, the
complexes of detector antibodies
and cTn I are captured to anti-cTn
I antibodies that have been
immobilized on test strip. Thus the
more cTn I antigens in blood
specimen, the more complexes
accumulated on test strip. Signal
intensity of fluorescence of
detector antibodies reflect the
amount of captured cTn I.
Shortened version:
The FineCare cTn I Rapid
Quantitative Test uses a sandwich
immunodetection method to detect
cTnI antigens in blood samples.

R1:
 Good’s buffer (pH 7.15)
 Sodium chloride (NaCl)
 Magnesium Chloride (MgCl2)
Reagents  a-Glucosidase
R2:
 Good’s buffer (pH 7.15)
 EPS-G7
R1:
 Good’s buffer (pH 8.0)
 Taurodesoxycholate
 Desoxycholate
 Calcium chloride
 Colipase (porcine)
R2:
 Tartrate buffer (pH 4.0)
 Taurodesoxycholate
 Color substrate
The test cartridge contains
fluorescence-labeled detector
antibodies and immobilized
capture antibodies. When a
sample is added, the detector
antibodies bind to cTnI antigens to
form complexes that migrate on
the nitrocellulose matrix and
accumulate on the test strip. The
intensity of the fluorescence signal
reflects the amount of cTnI
antigens in the sample.
Components
Material Provided
 Test cartridge in a sealed
R1:
pouch with dessicant
 N-Methyl-D-Glucamine (pH
 ID Chip
8.4)
 Detection buffer
 L-Lactate
 Pipette tip
 Leaflet with instructions for use
R2:
 NAD+
Material Required But Not
Provided
 FineCare FIA System
 Transfer Pipette Set (100 uL

 Reagents contain sodium
azide as preservative. Do not
swallow, avoid contact with
skin and mucous membranes.
 Reagent 1 contains animal and > P337+P313: If eye irritation
Warnings &
biological material. Handle the
Precautions
product as potentially
infectious.
 Saliva and skin contain a-
Amylases. Never pipette by
mouth and avoid skin contact.
size)
 Specimen collection containers
 Centrifuge (for serum/plasma
specimen only)
 Timer
 Reagent 2 Warning
> H319: Causes serious eye
irritation
> P280: Wear PPE
> P305+P351+P338: Rinse
cautiously with water for several
minutes. Remove contact lenses, if  Bring specimens to room
present and easy to do. Continue temperature before testing.
 Reagent 1 contains sodium
rinsing.  Frozen specimens must be
azide as preservative. Do not
completely thawed and mixed
swallow, avoid contact with
persists, get medical well prior to testing.
skin and mucous membranes.
advice/attention.  Specimens should not be
 Patients with gammopathy
 Reagent 1 contains sodium frozen and thawed repeatedly.
might give falsified results.
azide as preservative. Do not  Only clear, non-hemolytic
swallow, avoid contact with specimens can be used.
skin and mucous membranes.
 Reagent 1 contains animal
material. Handle the product
as potentially infectious.
 Many other clinical reagents
contain lipase or high
 concentration of detergents.
Avoid contamination and carry
over.

Conversion
U/L --> ukat/L: 0.0167 U/L --> ukat/L: 0.0167 U/L --> ukat/L: 0.0167
Factor:
SERUM/PLASMA
 Ascorbic acid
 Bilirubin (Conjugated and
unconjugated)
 Ascorbic acid
 Hemoglobin
 Ascorbic acid  Bilirubin (Conjugated and
 Triglycerides
 Bilirubin (Conjugated and unconjugated)
URINE
Interfering unconjugated)  Triglycerides
 Ascorbic acid
substances  Hemoglobin  Sulfapyridine
 Conjugated bilirubin
 Triglycerides  Sulfasalazine
 Boric acid
 N-acetylcystein (NAC)  Hemoglobin (interferes at low
 Glucose
concentrations)
 Hemoglobin
 Protein
 Sodium-Oxalate
 Urobilinogen
SERUM/PLASMA Conventional S.I.
Conventional S. I
Conventional S.I. (U/L) (ukat/L) Normal <0.30 ng/mL
Reference (U/L) (ukat/L)
(U/L) (ukat/L) Women <247 <4.12 At risk of AMI ≥0.30 ng/mL
Range ≤ 60 ≤ 1.00
Women Men <248 <4.14
<100 <1.67
/Men
 URINE
Conventional S.I.
(U/L) (ukat/L)
Women <447 <7.45
Men <491 <8.18
Step 1: Preparation
 Before testing, activate “use” in
setting then save it.
 Ensure that the lot number of
the Test Cartridge matches ID
Chip and Detection Buffer.
Insert ID Chip into Finecare
FIA System.

Test Step 2: Sampling

Procedure  Draw 75 uL of whole
blood/serum/plasma with a
transfer pipette and add into
the Detection Buffer tube

Step 3: Mixing
 Close the lid of Detection
Buffer tube and mix the
sample mixture thoroughly by
shaking it about 10 times.
Step 4: Loading
 Pipette 75 uL of sample
mixture and load it into the
sample well of the Test
Cartridge.

Step 5: Testing
Two Modes:
 Standard Test mode
- Insert the Test Cartridge onto the
holder right after adding sample
mixture to the sample well.
- Press “Test” to start testing.
 Quick Test mode
- Set the timer and count down
right after adding sample mixture
into the sample well and leave it at
room temperature for 15 minutes.
- Then, insert the Test Cartridge
onto the holder.
- Press “Test” to start testing.

 Results are displayed on main

screen or be printed by
pressing “Print”.
 Discard the used Test Catridge