Blakes Practical Two Flashcards
1
Q
Liquid GGT clinical significance on pointe scientific
A
-used in the diagnosis and treatment in liver disease
- such as Alcoholic cirrhosis, primary and secondary tumors
- Elevated GGT levels appear more pronounced in cases of obstructive jaundice and metastatic neoplasms.
2
Q
GGT reaction
A
L-y-Glutamyl-3-carboxy-4-nitroanilide + Glycylglycine ————–
3
Q
GGT reaction by word
A
GGT in the sample catalyzes the transfer of the glutamyl group from GLUPA- C to glycylglycine according to the above reaction. The amount of 5-amino- 2-nitrobenzoate formed is proportional to GGT activity and may be measured kinetically at 405nm.
4
Q
To prepare the working reagent then of GGT
A
Reagents are supplied as ready to use liquids. To prepare working reagent, mix 5 parts of R1 reagent with 1 part R2 reagent.
5
Q
Reagent storage and stability of GGT
A
Store reagents at 2-8°C. The reagents are stable until the expiration date if stored as directed. The working reagent is stable for 21 days at 2-8°C. NOTE: The R2 reagent is temperature sensitive and can be affected by prolonged exposure to room temperature. Return reagent to 2-8°C as soon as possible after use.
6
Q
reagent precautions GGT
A
- This reagent is for in vitro diagnostic use only.
- Do not use the reagent if the initial absorbance of the working reagent
is greater than 0.800 when measured at 405 nm against water or if the reagent fails to meet the stated parameters of performance - Do not pipette by mouth. Avoid ingestion and contact with skin as toxicity has not been established.
- Reagents in this kit contain sodium azide as a preservative. Sodium azide may form explosive compounds in metal drainlines. When disposing of reagents through plumbing fixtures, flush with copious amounts of water. For further information, refer to “Decontamination of Laboratory Sink Drains to Remove Azide Salts,” in the Manual Guide-Safety Management No. CSC-22 issued by the Centers for Disease Control, Atlanta, Georgia.
7
Q
GGT specimen collection and storage
A
- Use serum only. GGT activity is inhibited by most anticoagulants.
- It is recommended that specimen collection be carried out in accordance with NCCLS document M29-T2. No method can offer complete assurance that human blood samples will not transmit infection. Therefore, all blood
samples should be considered potentially infectious. - Serum GGT is reported stable in serum for up to seven days when stored at
2-25°C, up to one month when stored at 4°C, and up to one year at (-20°C)
and protected from evaporation.7 - All specimens and controls should be handled in accordance with good
laboratory practices using appropriate precautions as described in the CDC/NIH Manual, “Biosafety in Microbiological and Biomedical Laboratories
8
Q
GGT interferences
A
- Most anticoagulants used in blood collection tubes inhibit GGT activity.8
- Anti-epileptic drugs (phenytoin and barbituates) may falsely elevate GGT
levels.9,10 - Bilirubin to the level of 20 mg/dl has been found to exhibit negligible
interference (< 5%) in this assay. - Hemoglobin from 100-500 mg/dl has been found to show minimal depression
(approximately 5-7%) of recovered GGT activities.
NOTE: GGT level was 45 U/L for the bilirubin study and 48 U/L for the hemoglobin study.
9
Q
GGT expected values
A
Male: 8-37 U/L at 30°C, 9-54 U/L at 37°C
Female: 6-24 U/L at 30°C, 8-35 U/L at 37°C
10
Q
GGT linearity
A
Linearity: 0-800 U/L.
11
Q
LDH clinical significance
A
Increased levels of LD are associated with myocardial infarction. Levels
reach a maximum approximately 48 hours after the onset of pain and persist
about ten days. The degree of elevation is of value in assessing the extent
of damage and in developing a prognosis. LD elevations are also observed
in liver disease, pernicious anemia, in some cases of renal disease, and in
some cases of skeletal muscle trauma.
12
Q
LDH reaction and in words
A
L-Lactate + NAD+ with enzyme LDH————> Pyruvate + NADH + H+
Lactate dehydrogenase catalyzes the oxidation of lactate to pyruvate with
simultaneous reduction of NAD to NADH. The rate of NAD reduction can be
measured as an increase in absorbance at 340nm. This rate is directly
proportional to LD activity in serum.
13
Q
To prepare working reagent LDH
A
Reagents are supplied as ready to use liquids. To prepare a working
reagent, mix 5 parts Buffer (R1) with 1 part Co-enzyme (R2)
14
Q
Reagent storage and stability LDH
A
Reagent Storage and Stability
Reagents are stable until stated expiration if stored as directed. If a single
working reagent is required, prepare only the amount required to complete
current day’s testing. Discard remaining reagent after completion of testing.
Protect from light. Avoid microbial contamination
15
Q
LDH precautions
A
Precautions
1. This reagent is for in vitro diagnostic use only.
2. The reagents contain sodium azide (0.1%) as a preservative. Do not
ingest. Avoid skin and eye contact. Sodium azide may react with lead
and copper plumbing fixtures giving rise to explosive metal azides.
Flush with large volumes of water when disposing of the reagent.
3. All specimens and controls should be handled in accordance with good
laboratory practices using appropriate precautions as described in the
CDC/NIH
16
Q
LDH specimen and storage
A
- Non-hemolyzed serum is recommended. Red cells contain large
concentrations of LD. 5 - The serum should be removed from the clot promptly.
- Samples should be assayed soon after collection. LD in serum is reported
stable for two to three days at room temperature.9 - Do not freeze or expose the serum to high temperatures (37°C) as this may
inactivate thermolabile LD isoenzymes. 10 - Specimen collection should be carried out in accordance with NCCLS M29-
T2.11 No method can offer complete assurance that human blood samples
will not transmit infection. Therefore, all samples should be considered
potentially infectious
17
Q
Interferences of LDH
A
- Certain drugs and substances affect LD activity. See Young, et al. 12
- Bilirubin to the level of 20 mg/dl has been found to exhibit negligible
interference (≤ 5%) in this assay. - Hemolysis has been shown to significantly interfere with the assay at levels
as low as 100 mg/dl
18
Q
LDH linearity
A
Assay: 0-1000 U/L.
19
Q
LDH expected values
A
Male 50-166 U/L (30°C) 80-285 U/L (37°C)
Female 60-132 U/L (30°C) 103-227 U/L (37°C)
20
Q
GGT reference range and stability
A
range= 5-40U/L
Specimen stability: Best to maintain stability at RT, but can be stored for up to 14 days at refrigerated or frozen temperatures
21
Q
GGT primary role
A
Primary role is the detection and differential diagnosis of hepatobiliary disease.
22
Q
GGT examples of conditions where the highest elevations are
A
Examples of conditions where the highest elevations of GGT occur would be extra hepatic obstructions I.e. gallstones or a tumor blocking the bile duct (5 to 30X the upper reference range)
23
Q
Moderate GGT elevations
A
Moderate elevations can occur in injury caused by chronic alcoholism or drug ingestion
24
Q
GGT is useful in differentiating liver disease from what
A
Useful in differentiating liver disease from bone disease when it is elevated in the absence of an elevated ALP
25
Amylase clinical significance
The determination of amylase activity in serum is most commonly performed
for the diagnosis and treatment of diseases of the pancreas.
26
Amylase reaction
10 CNPG3 with enzyme a-amylase --------------------> 9 CNP + CNPG2 + 9G3 + G
-Amylase hydrolyzes the 2-chloro-p-nitrophenyl-
a-D-maltotrioside
(CNPG3) to release 2-chloro-nitrophenol and form 2-chloro-p-nitrophenyl-a-
D-maltoside (CNPG2), maltotriose (G3) and glucose (G). The rate of
increase in absorbance is measured at 405 nm and is proportional to the a-
amylase activity in the sample
27
A-amylase reagent storage
Do not use if:
1. The absorbance of the working reagent is greater than 0.600 when
measured at 405 nm against water in a cuvette with a 1 cm path length.
2. The reagent fails to meet stated parameters of performance.
3. The reagent is turbid or displays other evidence of bacterial contamination.
28
amylase interferences
1. A number of drugs and substances affect the determination of amylase.15,16
Young et al have published a comprehensive list of such substances.17
2. Macroamylase in the specimen can cause a measured hyperamylasemia,
that could lead to a false diagnosis of acute pancreatitis. However, no
clinical symptoms are usually associated with macroamylasemia.18
3. Bilirubin (30mg/dl) and hemoglobin (500mg/dl) have each been found to
have a negligible effect on this procedure.
4. Lipemic samples up to 1000 mg/dl have been reported to have no effect on
serum amylase determinations.1
29
Specimen Collection and Handling amylase
1. Unhemolyzed serum is the specimen of choice. Specimens should be
collected as per NCCLS document H4-A3.13
2. Anticoagulants, such as Citrate and EDTA, bind calcium that is needed for
amylase activity. Plasma with these anticoagulants should not be used.
3. Amylase in serum is reported stable for one week at room temperature (18-
25 °C) and for two months when stored refrigerated at 2-8 °C.
30
amylase expected values and linearity
Serum: 25-125 U/L
Linearity: 0-2,000 U/L
31
Amylase physiology
-Catalyzes the hydrolysis of alpha-glycosidic bonds in order to release various sugars from starches (releases sugars from starches)
-Produced in the acinar cells of the pancreas and salivary glands
32
amylase clinical significance
-Highest elevations occur in pancreatitis and inflammation of the salivary glands
-To diagnose acute pancreatitis, you should measure both amylase and lipase, Amylase peaks at 12 hours after onset and returns to normal after 3-4 days
33
Lab methods for amylase
-Laboratory Methods
-Couples enzyme reactions resulting in the measurement of NAD+ at 340nm
-Testing is available to measure the activities of p-amy and s-amy
34
Amylase reference range
27-131 IU/L
35
High levels of amylase
Acute pancreatitis (4-6X the upper limit of normal), pancreatic duct obstruction, pancreatic abscess, cholecystitis, mumps, intestinal obstruction, mesenteric thrombosis
36
Low levels of amylase
Cystic Fibrosis, Hepatitis, Cirrhosis
37
Alinity I side
* Alinity i-side:
* Immunoassay
* Thyroid studies
* Hepatitis
* Procalcitonin
* Bhcg, PSA, etc
38
Alinity I side bulk solutions.
Uses three bulk solutions.
Each bottle is labeled with the expiration date. Inventory tracking and consumable replacement are performed from the supplies screen.
39
Alinity I side bulk solutions.
Pre-trigger solution (1L bottle)
Trigger solution ( 1 L bottle)
Concentrated wash buffer (2-L bottle)
40
Pre trigger solution
A solution that contains 1.32% H2O2 solution that separates the acridinium dye from the conjugate that is bound to the microparticle to the microparticle complex. This action prepares the acridinium dye for the addition of Trigger solution.
Pre-Trigger solution is sensitive to light, is stored at a temperature of 2C to 8C, and is stable on the system for 16 days
41
Concentrated wash buffer
A solution that contains phosphate buffered saline and antimicrobial agents
This solution is diluted tenfold by the system and then is pumped to sample and reagent pipettor assemblies and to wash zones during assay processing
This solution is stored at a temperature of 15-30C and is stable on the system for 30 days
42
Trigger solution (1 L bottle)
A solution that contains 0.35N sodium hydroxide solution that produces the chemiluminescent reaction that provides the final read.
- This solution is stored at a temperature of 2-30C and is stable on the system for a maximum of 28 days
- Some assays require a shorter onboard stability period. For more information, see the assay documentation.
43
Reaction vessels
are disposable containers in which the CMIA reaction occurs
- The operator can add RVs at any time. A full bag of RVs, or partial bag of RVs, can be added into the RV hopper. Do not overfill the RV hopper.
- Inventory tracking and consumables replacement is performed from the supplies screen.
44
Processing center
is the main activity area of the processing module. samples and reagents are dispensed and mixed in reaction vessels (RVs) in the process and pretreatment paths where assay processing is preformed.
45
Processing center path
1.) Process path
2.) Pretreatment path
3. pipetting hardware
4. RV loader
46
Process path
The process path is covered, circular path that provides incubation at a controlled temperature, liquid aspiration, and wash points as necessary for assay processing
47
Pipetting hardware
-Pipettors detect, aspirate, transfer, and dispense samples and reagents into reaction vessels (RVs).
-There are three pipettors and active wash cups in the Alinity I processing center.
1.) sample pipettors
2.) Reagent pipettors
3.) Wash cups
48
Process path hardware
Components located around the process path cover are used to perform the assay processing activities
49
Pretreatment path
The pretreatment path is a covered, circular path that provides incubation at a controlled temperature for pretreatment assay protocols
- After the pretreatment assay protocol is completed, the sample is transferred to the process path.
50
the three components on the pretreatment path that are used to perform assay processing activities
1.) Pretreatment path motor
2.) Pretreatment vortexer
3.) Pretreatment Unload diverter
51
CMIA reaction process
Chemiluminescent micro-particle immunoassay is a detection technology used to measure analyte concentration Chemiluminescent microparticle immunoassay detection technology is used to determine the pressence of antigens, antibodies, and analytes in a sample
52
CMIA reaction sequence
is the order of interactions between the analyte in the sample and the reactants. The sequence is specific to the assay protocol
53
CMIA process steps
1. The sample and the paramagnetic microparticles coated with capture molecules are dispensed into the reaction vessel (RV). The vortexer mixes the reaction mixture.
2. The reaction mixture incubates. The analyte in the sample binds to the capture molecules on the paramagnetic microparticles and forms an immune complex.
3. A magnet attracts the paramagnetic microparticles (which are bound to the specific analyte) to a wall of the RV. The wash zone assembly washes the reaction mixture to remove unbound materials. Additional assay processing can now occur.
4. The pipettor dispenses a chemiluminescent, acridinium-labeled conjugate into the RV. The conjugate binds to the immune complex to complete the reaction mixture. The vortexer mixes the reaction mixture.
54
CMIA step two reaction
55
Consumables C-series
-The Alinity c-series uses three bulk solutions,
-Bulk solutions are liquid solutions that are provided in large quantities for use during sample, processing.
-Each bulk solution bottle is loaded on the bulk solution door.
Each bottle is labeled with the expiration date. inventory träcking and consumable replacement is performed from the Supplies Screen.
56
Alinity C-side bulk reagents: ICT reference solution ( 975 mL in a 1 L bottle)
-A midconcentration standard solution that is aspirated and analyzed by the ICT module before and after each sample. The solution provides a reference potential that is used in result calculation.
-ICT Reference Solution is stored at a temperature of 15°C to 30°C
and is stable on the system for 90 days.
57
Alinity C-side bulk reagents: Acid wash (0.5L bottle)
-An acidic wash solution that is used by the cuvette washer to clean the cuvettes after sample analysis.
-Acid Wash is stored at a temperature of 15°C to 30°C and is stable on the system for 30 days.
-CAUTION: Follow all safety precautions as described on product-specific labels, in
the product documentation, and in product-specific Safety Data Sheets.
58
Alinity C-side bulk reagents: Alkaline wash (0.5L bottle)
-An alkaline wash solution that is used by the cuvette washer to clean the cuvettes after sample analysis.
-Alkaline Wash is stored at a temperature of 15°C to 30°C and is stable on the system for 30 days.
-CAUTION: Follow all safety precautions as described on product-specific labels, the product documentation, and in product-specific Safety Data Sheets
59
Reaction carousel hardware
The reaction carousel hardware components position the cuvettes for sample and reagent dispense, mixing. photometric or potentiometric analysis, and cuvette washing.
1. Mixers
2. ICT unit
3. Lamp
4. ICT high-concentration waste area
5. Cuvette washer
6. Water bath overflow and waste area
7. Cuvette segments
8. Reaction carousel
9. High-concentration waste pump
60
Processing centers
The processing center is the main activity area of the processing module. Samples and reagents are dispensed and mixed in a reaction carousel where assay processing is performed.
1. Pipetting hardware: Aspirates and dispenses sample and reagents
2. Reaction carousel hardware: Positions the cuvettes for sample and reagent dispense, mixing, photometric or potentiometric analysis, and cuvette washing
61
Pipettors
Pipettors detect, aspirate, transfer, and dispense samples and reagents into cuvettes.
There are three pipettors:
1. Sample pipettor
2. Reagent 1 pipettor
3. Reagent 2 pipettor
62
Reaction carousel
-Samples and reagents are dispensed and mixed in a reaction carousel where assay processing is performed.
-The reaction carousel has 17 cuvette segments surrounded by a 37°C water bath. Each cuvette
I segment contains 11 cuvettes for a total of 187 cuvettes.
28
63
Cuvette washer
The cuvette washer is a device with eight nozzles that clean and dry each cuvette before and after each cuvette is used.
64
High concentration waste pump
The high concentration waste pump works with the cuvette washer to aspirate waste from the cuvettes to the optional high-concentration waste bottle or the drain.
65
Photometric measurements
-Photometric technology is the measurement of the amount of light that a sample absorbs.
-Beer's Law establishes the mathematical relationship between the absorbance of the solution and the concentration of the analyte.
-The optical system on the processing module is a direct photometry system that directs and aligns the light from the source lamp, through the water bath and the cuvette, to the optics unit.
66
The optical system
The absorbance of the solution changes as
-The optical system directs and aligns only the light that originates from the source lamp and simultaneously measures the intensity of 16 different wavelengths.
-The absorbance of the solution changes as the reaction progresses. Measurements occur either when all the reactant is depleted and the reaction is stable (end-point assays) or when the reactant reaches a stable rate (rate assays).
67
The C-series uses
The c-series uses the optical measurement to obtain absorbance readings and then converts them to assay-specific analyte concentration units or assay-specific qualitative interpretations.
68
Potentiometic measurements
-The c-series uses the potentiometric detection technology to measure the electrical potential in a sample.
-Integrated Chip Technology (ICT) methodology uses solid-state, ion-selective electrodes that are contained in one chip (ICT module) to measure sodium (Na+), potassium (K*) and chloride (CI-).
69
ICT reference solutions
- ICT Reference Solution and ICT samples are delivered to the ICT module where the following measurements are captured:
* The potential difference between the sample and the ICT Reference Solution for each electrode (reference, Cl-, K+, and Nat).
* The potentiai of each electrode in contact with the ICT Reference Solution
* The potential of each electrode in contact with the sample
The ICT measurements are obtained in millivolt readings that are used by a data reduction calculation to calculate the final result concentration.
(el The final millivolt readings are converted to assay-specific analyte conversion units for reporting.
70
Mixers
The Alinity c processing module has two mixers that mix sample with reagent in the cuvette. After each mixing operation, the exterior of the mixer is washed in the wash cup located beneath the mixer.
71
ICT unit
The integrated chip technology (ICT) unit has an
ICT probe and an ICT module. It is used to perform the indirect potentiometric analysis of sodium (Nat), potassium (K+), and chloride (CI).
72
Tumor or neoplasm
Abnormal/uncontrolled proliferation of cells.
73
Benign tumor
Tumor that remains confined to its primary site.
74
Malignant tumor
Tumor that is capable of invading surrounding
normal tissue and metastasizing (spreading) through
the circulatory and lymphatic systems to distant
body sites—also called “Cancer”.
75
Cancer
a malignant neoplasm or tumor
76
Tumor marker
a substance synthesized by the tumor or by the
host in response to a tumor that can be used to
detect the presence of the tumor
77
Desirable Characteristics of a tumor
marker
* Specific for cancer
* Always present with tumor
* Amount of marker produced should correlate well with the
tumor load
* The half-life of the marker should be short so serum levels
must drop producing undetectable concentrations when
patient is in remission.
* Levels of marker should have prognostic value
* Assay for the marker should be analytically sensitive, specific,
accurate, precise, easy to perform, inexpensive, and rapid.
78
Uses of tumor markers
* Screening for disease
* Aid in diagnosis for symptomatic patients
* Aid in clinical staging
* Measurement of tumor burden
* Monitoring response to therapy
* Detecting recurrence of disease
* Prognostic indicator
79
Types of tumor markers
* Enzymes and isoenzymes
* Hormones, neurotransmitters and their metabolites
* Receptors (estrogen, progesterone, androgens,
corticosteroids)
* Proteins (immunoglobulins, glycoproteins,
carcinoembryonic proteins or oncofetal antigens)
* Genetic markers (Oncogenes and suppressor genes)
* Other markers (amino acids)
80
Carcinoembryonic antigen (CEA)
* Belongs to the family of glycoproteins.
* It is normally produced during fetal development, but
the production of CEA stops before birth.
– It is not usually present in the blood of healthy adults
81
CEA Elevated in
– Colon cancer
– Lung cancer
– Gastric cancer
– Breast cancer
– Pancreatic cancer
– Ovarian cancer
– Uterine cancer
82
CEA is the most
* CEA is the most widely used tumor marker for
colorectal cancer
83
Prostate Specific Antigen (PSA)
Serine protease produced exclusively by the epithelial cells in
the prostate.
* PSA is the only tissue-specific marker identified so far, but is
not specific for prostate cancer.
– Found in small amount in normal prostate
– Also elevated in Benign Prostatic Hyperplasia (BPH)
* Only tumor marker recommended for screening for prostate
cancer and is a useful diagnostic tool.
* PSA is used to screen, stage, monitor treatment, and
recurrence of prostate cancer.
84
PSA exist in what form
what type of test
* PSA exists in both free and complex (e.g. PSA
complexed to alpha1-antichymotrypsin)form in
serum.
* Immunoassays are available for total, free and
complex PSA.
* Measuring the free PSA percentage (free/total x 100)
or free PSA to PSA-ACT ratio may to differentiate b/w
Prostate cancer and BPH.
85
Human Chorionic Gonadotropin (HCG)
* Pregnancy associated hormone, normally secreted by the
synctiotrophoblastic cells of the placenta.
– Dimer composed of alpha and beta subunits
* Alpha subunit is found in FSH, LH, and TSH
* Beta subunit is specific to hCG
* hCG is commonly measured to confirm pregnancy
* hCG is also a useful marker for tumors of the placenta
(trophoblastic tumors), and germ cell tumors of the
ovaries and testes.
86
Alpha-Fetoprotein (AFP)
* AFP is a glycoprotein that is normally produced by the fetal
yolk sac and fetal hepatocytes during embryonic
development.
* Structurally related to Albumin
* Production of AFP declines rapidly after birth and healthy
adults have negligible/undetectable levels in serum.
* Can be used during maternal serum screening for neural
tube defects and for Down syndrome.
87
AFP most useful what
may be elevated during
combination of
* AFP is the most useful serum marker for
diagnosis and management of hepatocellular
carcinoma (HCC).
* May also be elevated during pregnancy and in
cases of benign liver diseases (e.g. hepatitis)
* The combination of hCG and AFP is used to
classify and stage germ cell tumors.
88
Estrogen Receptor (ER)
* ER is a 70 kDa protein found in the nuclei of mammary and
uterine tissues.
* Transcription factor.
* Measurement of ER/PR in breast tumor cytosol is used to ID
those pt’s who will MOST likely benefit from hormone therapy.
* Approximately 55-60% of pt’s whose primary tumor
demonstrated ER will respond to hormone therapy.
89
Estrogen Receptor (ER)
* ER is a 70 kDa protein found in the nuclei of mammary and
uterine tissues.
* Transcription factor.
* Measurement of ER/PR in breast tumor cytosol is used to ID
those pt’s who will MOST likely benefit from hormone therapy.
* Approximately 55-60% of pt’s whose primary tumor
demonstrated ER will respond to hormone therapy.
90
CA 125
* Glycoprotein expressed by epithelial ovarian tumors and
other pathological and normal tissues of mullerian duct
origin.
* CA 125 is most useful as a marker for ovarian cancer
– However, elevation is seen in a number of nonovarian
cancers (not specific)
* Useful in monitoring treatment of disease in ovarian
cancer patients.
91
CA 19-9
* Blood group antigen.
* Produced by normal pancreatic and biliary ductular cells and
by colon, endometrial, gastric and salivary epithelia.
* Marker for colorectal and pancreatic cancer.
* Approved by FDA to monitor patients with pancreatic cancer.
* May also be increased in diseases associated with biliary tract
obstruction and pancreatitis.
92
CMIA step two reaction
5. The wash zone assembly washes the reaction mixture to remove unbound materials.
6. The Pre-Trigger Solution nozzle dispenses the Pre-Trigger Solution (hydrogen peroxide) into the reaction mixture. The vortexer mixes the reaction mixture. The Pre-Trigger Solution:
Creates an acidic environment to prevent the early release of energy (light emission
Helps to prevent any clumping in microparticles
*Separates the acridinium dye from the conjugate that is bound to the microparticle complex. This action prepares the acridinium dye for the next step.
The CIA optical system performs a background read.
7. The Trigger Solution nozzle dispenses the Trigger Solution (sodium hydroxide) into the reaction mixture. The Trigger Solution creates an alkaline environment that, with the exposure to peroxide in the Pre-Trigger Solution, causes the acridinium dye to undergo an oxidative reaction. The oxidative reaction causes a chemiluminescent reaction to occur. N-methylacridone forms and releases energy (light emission) as N-methylacridone returns to its ground state.
The CMIA optical systern measures the chemiluminescent emission (activated read) over a predefined time period to determine a result.
93
Tumor markers can be measured by
be detected by 1) chemical, 2)immunologic, or 3) molecular techniques
“Early detection, better chance of cure”
94
ALP: tumor marker
-Quantification: helpful in evaluating metastatic cancer with bone or liver involvement
–increased Osteoblastic lesion [prostatic cancer with bone metastasis]
– Increased Osteoclastic lesion [breast cancer with bone metastasis]
– with increased 5’-NT or γ-GTT, elevated ALP is of liver origin
– PALP: 1st identified Regan isoenzyme (oncodevelopmental marker)
95
LD non specific tumor marker
-Variety of cancers; liver cancer, Hodgkin’s lymphoma, acute leukemia, germ cell testicular
cancer (non-seminoma), seminoma, neuroblastoma, other carcinoma (breast, lung..)
– Serum LD correlated with tumor mass (solid)
– Serum value rarely considered from tumor staging, but testicular cancer and melanoma.
E NZYMES
96
Hepatobiliary Diseases
-Biliary tree obstruction induces the synthesis fo ALP by hepatocytes.
* About three fold elevation of its activity by extrahepaic obstruction than intrahepatic.
* Infectious hepatitis: mild increase or normal ALP activity.
* Drug theraphy could incread ALP activity.
97
Bone Diseases
Bone ALP produced by osteoblast
* Hypophosphatasia: severe bone disease and impaired bone growth
* Paget disease (osteitis deformans)
* Vit D deficiency
* Osteoporesis
* Bone cancer
* Primary and secondary hyperparathyroidism
98
Paget disease (osteitis deformans)
what enzyme is involved
ALP
A chronic disorder that results in enlarged and deformed bones (also known as osteitis deformans).
99
LD is found in
Invariably found only in the cytoplasm of the cell
* Increased levels are found in hepatic, cardiac, skeletal muscle and in several
hematological disorders
In hematological (like hemolytic anemia and megalobastic anemia) and oncology
(liver cancer germcell tumor (LD-1), LD monitoring is important.
100
Neuron Specific Enolase
-Found in neuronal tissue and cells in neuroendocrine system.
[Cancer originated in the neuroendocrine system]
– Released into the blood circulation because of cell lysis, not secretion.
– NSE also released into CSF with neuronal injury
– SCLC, neuroblastoma, pheochromocytoma, carcinoid, medullary carcinoma of the
thyroid, melanoma and pancreatic endocrine tumor
101
PROSTATIC ACID PHOSPHATASE (PAP)
not as sensitive as PSA for detection of early cancer
– Still important for the patient who does not secrete PSA
– Predicting recurrence after radical prostatectomy
102
PSA Prostate specific antigen assay
* Sandwich immune assays:
* Different assay or even same assay with different reagents
* PSA exists in both free & complex form in serum. (e.g. PSA complexed to ACT (alpha1-
antichymotrypsin))
* Immunoassays are available for total, free and complex PSA.
* Measuring the free PSA percentage (free/total x 100) or free PSA to PSA-ACT ratio may to
differentiate b/w Prostate cancer and BPH.
103
T HE UROKINASE- PLASMINOGEN ACTIVATOR (UPA) SYSTEM
Inactive single polypeptide! cleavage (b/w K158 & I159) ! Active form [A chain
(interact with cell surface receptor)] [B chain (catalytic active)]
Plasminogen ! --> plasmin --> degrade ECM & activate MMPs
104
Measured its catalytic activity (original) replaced by ELISA
or ER status and equally powerful as nodal status
Breast cancer, colorectal cancer. [may be for general cancer]
105
Cathpsins
Lysosomal protease enzyme
Cathepsin 1) B, 2) D, 3) L
– Synthesized as large molecule precursor
– Addition to protease activity, release growth factors
106
CB: activate
Activate uPA & MMPs
– Expression and location altered in tumors relative to normal tissue
– Increased expression: 1) breast, 2)colorectal, 3) gastric, 4) lung, 5) prostate carcinomas, 6) glioma,
7) melanoma, 8) osteroclastomas
– Altered location: 1) colon carcinoma, 2) thyroid cancer, 3) glioma, 4) breast epithelial tumor
