25/26 - Basic Clinical LABS Flashcards
(100 cards)
1
Q
Why are Labs Ordered?
A
To Confirm a DIAGNOSIS
or to distinguish amoung different forms of a disease/pathology
To assess the severity of a condition
To monitor progress / therapy
To detect side-effects / complications
2
Q
Typical Labs ordered in an ANNUAL CHECKUP
A
CBC
Lipid + Glucose Panel
Liver Fxn Panel
Blood tests for kidney fxn
Urinalysis
Thyroid / C-reactive Protein / HbA1C
Vitamin D / Homocysteine / PSA
3
Q
CBC
Typical Lab ordered for Annual Checkup
A
Complete Blood Count
includes:
WBC / RBC
WBC differential
Hematocrit / Hemogloblin
Platelet Count
4
Q
Lipid + Glucose Panel
Typical Lab ordered for Annual Checkup
A
Total Cholesterol
LDL + HDL
LDL/HDL Ratio
Glucose
5
Q
Liver (Hepatic) Function Panel
Typical Lab ordered for Annual Checkup
A
blood levels of:
Total Protein
Albumin / Bilirubin
Liver Enzymes
6
Q
Blood tests for KIDNEY FUNCTION
Typical Lab ordered for Annual Checkup
A
Serum CREATININE
- *BUN**
- *blood urea nitrogen**
7
Q
Urinalysis
Typical Lab ordered for Annual Checkup
A
Examine specimen for:
color / clarity / odor / pH / spgr
protein / glucose / ketones
Also microscopic analysis for:
bacteria / crystals / blood cells
8
Q
Assays for VITAMIN status Focus on:
A
DIRECT measurement of the
vitamin / cofactor / precursor
in biological fluids or blood cells
or measurement of:
Urinary Metabolites
Biochemical Function
that requires the vitamin / cofactor
ex. G6P for thiamine deficiency
9
Q
Vitamin A
How is it clinically assayed?
A
Fat Soluble
Vision / growth / Reproduction
night blindness
Fluorometric
HPLC / RIA
Photometric
10
Q
Fat Soluble Vitamins
How are they clinically assayed?
A
(ADEK)
HPLC for ALL of them
- RIA** for all except *vitamin E
- not used much anymore due to cost / hazard / license*
Photometric for all except vitamin D
11
Q
Vitamin D
How is it clinically assayed?
A
Fat soluble vitamin
Ca2+ metabolism / bones / teeth
rickets / osteomalacia
Competitive Binding Protein = CPB
HPLC / RIA
no photometric assay
12
Q
Vitamin E
How is it clinically assayed?
A
Fat-soluble Vitamin
Antioxidant for unsat-lipids
lipid peroxidation / fragile RBC
Erythrocyte Hemolysis
HPLC, NOT RIA
Photometric
13
Q
Vitamin K
How is it clinically assayed?
A
Fat-Soluble Vitamin
blood clotting / osteocalcins
increased clotting time + hemorrhages
ProThrombin Time
HPLC / RIA
Photometric
14
Q
Thiamine
Water Soluble Vitamin, how is it CLINICALLY ASSAYED?
A
- *B1**
- beri beri*
HPLC / Fluorometric / microbial
Transketolase
15
Q
Riboflavin
Water Soluble Vitamin, how is it CLINICALLY ASSAYED?
A
B2
redox / dermatitis, photophobia
Enzyme
HPLC / Fluorometric / Microbial
16
Q
Pyridoxine
Water Soluble Vitamin, how is it CLINICALLY ASSAYED?
A
B6
AA + phospholipid metabolism / anemia + convulsions
Tyrosine Decarboxylase
HPLC / no fluoro / Microbial
17
Q
Niacin
Water Soluble Vitamin, how is it CLINICALLY ASSAYED?
A
B3
REDOX / pellagra
Fluorometric / Microbial
no HPLC, too common in the immune system
18
Q
Folate
Water Soluble Vitamin, how is it CLINICALLY ASSAYED?
A
B9
synthesis of Nucleic+Amino Acids / megaloblastic ANEMIA
same assays as B12
IMMUNOASSAY
CPB = competitive protein binding
no HPLC, common in immune system / no fluoro
Microbial
19
Q
Cyanobalamin
Water Soluble Vitamin, how is it CLINICALLY ASSAYED?
A
B12
AA + Lipid metabolism / Pernicious+megaloblastic ANEMIA
same assays as B9, folate
IMMUNOASSAY
CPB = competitive protein binding
- *Microbial**
- no HPLC, common in immune system / no fluoro*
20
Q
Biotin
Water Soluble Vitamin, how is it CLINICALLY ASSAYED?
A
B7
Carboxylation / dermatitis
AVIDIN BINDING
Enzymes
PHOTOmetric
Microbial
21
Q
Pantothenic Acid
Water Soluble Vitamin, how is it CLINICALLY ASSAYED?
A
B5
Central metabolism / burning feet syndrome
Enzymes
CPB
PHOTOmetric
Microbial
22
Q
Ascorbate
Water Soluble Vitamin, how is it CLINICALLY ASSAYED?
A
C
connective tissue / scurvy
PHOTOmetric
HPLC
23
Q
What Vitamin can be analyzed by
Erythrocyte Hemolysis?
A
Vitamin E
antioxidant / lipid peroxidation
24
Q
Which Vitamin can be analyzed by
PROTHROMBIN TIME?
A
Vitamin K
blood clotting / hemorrhages
25
**Which vitamins can be analyzed by**
**IMMUNOASSAY?**
**_FOLATE + Cyanobalamin_**
B9 / B12
both also CPB & Microbial assays
26
**Which vitamin can be analyzed by**
**AVIDIN BINDING?**
**_BIOTIN_**
B7
carboxylation / *dermatitis*
27
**Oral Glucose Tolerance Test**
What is done **prior / during the test?**
**OGTT**
**Controlled diet** for **3 days prior** to test
After an **overnight fast**, patient is to swallow a solution with
**_75 grams of glucose_**
Then **blood samples** are drawn at regular intervals and
**assayed for glucose**
28
**Oral Glucose Tolerance Test**
**What does it measure?**
What is a **normal response?**
Use a **_BLOOD DRAW_** to measure
**_Glucose concentration over time_**, after the OGTT load
**_Normal Response_**
**Peak @30 min**,
return to **fasting @2hours**
fasting = 700-1050 mg/L
29
**OGTT**
What is a **Diabetic/abnormal Response?**
What values indicates **Diabetes?**
in an abnormal diabetic response,
BOTH **Starting Glucose** & **Glucose PEAK** is **HIGHER**
***_slow return to fasting level_***,
or not dropping to NORMAL fasting (700-1050mg/L)
* *_DIABETES_**
* *_\>_ 2000 mg/L**, after **2 hours** after the OGTT load
30
**Main differences in clinical test results for**
**Diabetic vs Non-diabetic**
* *_DIABETES_**
* *_\>_** **2000 mg/L, 2hours post OGTT**
* *LATER & HIGHER Peak**
* *Fasting level is also HIGHER**
**_Normal / *Non-diabetic*:_**
**Peak @ 30 min**
Return to **fasting @ 2 hours**, (normal = 700-1050 mg/L)
31
**Why do we NOT use urinary glucose levels?**
Why is BLOOD GLUCOSE preferred?
The fasting blood glucose level can be measured easily with **little interference**, and it **correlates well with the severity of the disease**.
In healthy individuals, _Glucose MUST **exceed \>1800 mg/L**_
before excessive glucose **appears in the urine**
In **Diabetic** patients, the renal threshhold can increase to **\>3000mg/L**
**_Urinary glucose is NOT a sensitive marker_**
* Also, excessive glucose excretion can be caused by:*
* *Pregnancy / Rickets / Osteomalacia**
32
What is often shown in **PREDIABETES**?
* *_IFG**_ and/or _**IGT_**
* AT RISK for **_T2DM_*** **_+_** ***_stroke + heart attacks_***
## Footnote
**_IMPAIRED FASTING GLUCOSE_**
FBG = **1000-1250 mg/L**
*\>1260 mg/L is diabetes*
**_IMPAIRED GLUCOSE TOLERANCE_**
blood glucose level is **1400-2000 mg/L**
AFTER a **2-hour OGTT**
*\>2000 is diabetes*
33
**3 Ways to DIAGNOSE DM by blood glucose levels**
What levels?
**_Tests should be CONFIRMED on a SUBSEQUENT DAY_**
1) **FPG \> 1260** mg/L
* (prediabetes = 1000-1250)*
2) Casual plasma **B****G****\> 2000** mg/L
3) **2hr OGTT** w/ post load value **\> 2000** mg/L
34
**What are the 3 Ketone Bodies?**
involved in KETOSIS
**Acetone**
**_B-hydroxybutyrate_**
**_Acetoacetate_**
**_Organic acids_**, if their levels rise TOO HIGH --\> **blood pH = ACIDIC**
35
**At what LEVELS is KETOSIS seen?**
What **symptoms** are seen?
Seen with levels of **_30-70_** mmol/L
**Acetone** on the **breath** may be detectible
36
**When does Acidotic Ketosis occur?**
when the **_Blood pH DROPS BELOW 7.3_**
**B-hydroxbutyrate + acetoacetate**
Organic acid, concentrations are TOO HIGH
overcome the **buffering action of the serum**
37
**What are the 2 TYPES of**
**BLOOD GLUCOSE METERS?**
and what do they **measure?**
FIrst we need to get a **single drop of blood**, from **FINGERSTICK**
**_Electrochemical Glucose monitor_**
uses glucose oxidase -\> **hydrogen peroxide** -\> o2
**small electrical current**
**_Photometric Glucose Monitor_**
similar to above, but **color change is measured**
38
**What does HbA1c measure?**
And how is it **correlated with Diabetes?**
_**AVERAGE BG** over the last **2-3 Months**_
**HIGH HbA1c :** directly proportional to : **degree of HYPERglycemia**
indicates a *_loss of BG control or pt's compliance_*
assayed by:
**electrophoresis
ion exchange / afffinity chromatography**
39
**What is NITROGEN BALANCE**?
24-hour urine collection used to measure
What are its **Positive / Negative values?**
**_Dietary Nitrogen INTAKE_** - ***_Excretion or Losses_***
(mainly from AA's in protein) - (mainly from URINE)
**Positive Value** = **Growth + Development + Pregnancy**
* **_Negative Value = INCREASED catabolism of AA's + Purines_***
* may call for dietary intervention*
40
**Where are AA's filtered out from?**
**What occurs if the transport is saturated or defective?**
AA's are filtered in the **_KIDNEY_**,
*reabsorbed in renal tubules by* **saturable/active transport systems**
If this system is ***_saturated or defective_***,
the **AA's** are left in the urine and are **excreted**
**_AMINOACIDURIA_**
results
41
**What causes PRIMARY Acidurias?**
**_MUTATIONS IN ENZYMES_**
that are involved in **AA metabolism**
*loss in activity of enzyme* --\> RAISES concentration of **precursors**
which will appear in the **urine**
42
**What is SECONDARY Aciduria?**
**_DAMAGE TO KIDNEY_**
from *various causes:*
**Viral infection** / **APAP toxicity** / **Rickets**
Genetic defects in **enzyme/transporters in kidney**
43
**What are SCREENING TESTS for**
**Aminoacidurias?**
**_Screening Tests_**
cheap & fast, *not quantitative*
specimen can come from _urine / blood / **CSF**_
**TLC w/ ninhydrin spray**
**Photometric** (UV absorbance)
**Guthrie** (microbiological)
**FeCl3** test for PKU
44
**What are QUANTITATIVE TESTS**
**for AminoAcidurias?**
**_Quantitiative Tests_**
*slower, not so cheap + needs **special equipment***
may use _urine or blood_ w/ pre-treatment
**_HPLC + Ion-Exchange Chromatography_**
detection by **UV / MS** for liquid chromatographic methods
**_Capillary Electrophoresis_**
UV / MS detection
45
**OTHER Organic ACIDURIAS**
other secondary metabolites
Diabetes & Ketone bodies = Acetoacetate + B-hydroxybutyrate
Defects in **propionate metabolism**
**B12** *deficiency* -\> HIGH levels of **propionate + methylalonate**
* *_MSUD_** = maple syrup urine disease
* defect in AA metabolism* -\> high levels of certain KETO ACIDS
46
**What type of Analysis/Test is done for**
**VOLATILE ANYLYTES?**
**_GC / MC_**
for small **volatile analytes = you can SMELL IT**
_Ex. Maple Syrup - MSUD_
*_NOT used for amino Acids_*
47
**What are anlytes for KIDNEY FUNCTION TESTS?**
Where do the **specimens come from?**
**_URINE / BLOOD / SERUM_**
Analytes:
**UREA / Uric Acid + Urate / AMMONIA
CREATINE + CREATININE**
High capacity of kidney for filtration -\> waste products are **not seen**
until **renal function is GREATLY compromised**:
Major function of kidney is elimination of **nitrogenous wastes**
urea / ammonia / urate
48
**What levels/tests are used to monitor the**
**filtration ability of the KIDNEY?**
**_BUN**_ (blood urea nitrogen) & _**URINARY CREATININE_**
Nitrogen:Creatinine Ratio is typically **10:1** or **20:1**
HIGH RATIO = *decreased renal perfusion*
cardiac failure / GI bleeding / excess protein / drugs
*Low Ratio does not indicate pathology*
found with *low protein intake*
but also with SEVERE LIVER DISEASE
49
**What ORGAN(S) do UREA LEVELS track the health of?**
**BOTH**
**_LIVER + KIDNEY_**
Synthesis is in the liver
Excretion is through the kidney
**Urea is the MAJOR route by which we excrete NITROGEN**
50
**How do we clinically ASSAY**
**UREA?**
**Urea** + urease -\> **Ammonia:**
**_Berthelot_**
ammonia + phonol -\> **measure absorbance @ 560nm**
* *_Glutamate dehydrogenase_**
* *UV absorbance @ 340nm**
Alternate CHEMICAL method
*DIRECT reaction with* ***_diacetyl --\> DIAZINE_***
Buttery smell = diacetyl
51
**What are physiological sources of**
**AMMONIA?**
**_GI TRACT_** is the main source
NH3 is generated by **bacterial action
(**proteases / ureases / amine oxidases)
* **Some is made by the _URIC ACID CYCLE_***
* unless there is a defect*
52
**HYPER-Ammonemia**
Causes / Effects
Caused by:
* *Inherited Deficiencies** in **UREA CYCLE ENZYMES**
* *Liver Disease** or **Renal Failure**
* though the MAJOR source of ammonia is the GI TRACT*
High Ammonia = **Toxic, depletes TCA cycle** & reduce energy
**Nerve Cell damage** -\> **Nausea / Coma / Death**
53
**How is _Ammonia_ Analyzed / tested for?**
**_SERUM_** samples are preferred
Quantitative Methods:
* *_Glutamate Dehydrogenase_** = PREFERRED
* *Berthelot Method**
**Special precautions to *avoid CONTAMINATION*** from:
**poor ventilation** / **smoking residue**
poor venipuncture technique
*delays in specimin analysis --\> **degradation***
54
**Creatine vs CreaTINine**
**Creatine --\> CreaTINine**
SPONTANEOUSLY CONVERTED at a constat rate in the **_Muscle_**
reflects the amount of muscle tissue
Creatine **+ Creatine Kinase -\> Phosphocreatine**
**_Phosphocreatine is also converted to CreaTINine_**
imortant in restoring ENERGY
55
What does **CreaTINine clearance reflect?**
**What organ?**
**CrCl**
Reflects the **ability of the KIDNEY** to
## Footnote
**FILTER metabolic byproducts from the BLOOD**
56
**How is Creatine / CreaTINine Analyzed/Assayed?**
Specimen = **_Serum / Plasma_**
CreaTINine reacts w/ alkaline picrate --\> **orange-red complex**
measured **_spectrophometrically_**
Also enzymatic methods, creaTINine -\> creatine -\>
**_color change or UV abosorbance change_**
or **_algorithim to compare with measured GFR_**
24 hour urine collection is ideal, but 12 is fine
57
How is **Uric Acid (Urate) produced?**
what does it do?
* *Catabolism of _PURINES_** = Adenosine & Guanine
* minor source is from DIETARY PURINES*
**Uric Acid** is excreted through **kidney --\> urine**
in VIVO it is an **antioxidant**, but will **precipitate \<5.6 pH**
**_GOUT_**
poor circulation etc.
58
What is **HYPER-Uricemia**?
What **diseases does it cause?**
Serum / plasma level of urate **\> 7.0** mg/dL for men **\>6.0** for women
**_GOUT_**
due to urate **precipitation in JOINTS**
**_Lesch-Hyhan Syndrome_**
*deficiency in HGPRT enzyme & blocks purine solvage*
HIGH levels of **hypoxanthine + guanine --\> hyperuricemia**
59
**How is Urate/Uric Acid Clinically ASSAYED?**
**_HPLC_**
**_Phosphotungstic Acid = PTA_**
BLUE color as PTA is reduced by urate
**_Uricase_**
UV absorbance as enzyme, urea -\> allotoin
***Dry chemistry systems***
60
**Sketch the transport of LIPIDS**
**throughout the BODY**
61
**What is the role of Lipoproteins?**
Lipoproteins **_transport LIPIDS through circulation_**
in **micelllular-like complexes**
specialized system of lipoproteins & receptors
Body fat is mainly composed of
**triglycerides** = major **long term energy storage**
TG / Cholesterol form fat droplets
62
**What are LDLs?**
***_Low Density Lipoprotein_***
ideally **\<100mg/dL**
Still a **necessary part of LIPID TRANSPORT**
LDL-\> peripheral tissue
Associated with **Atherosclerosis** = HARDENING of arteries
**fat deposits** in the lining of arteries --\> **CHD** or artery blockage
63
**What are HDL's?**
**_HIGH density Lipoprotein_**
ideally **\>40 mg/dL**
**Aids in the REMOVAL OF CHOLESTEROL**
from _arterial wall plaques_
LDL -\> Peripheral Tissue -\> HDL -\> back to LIVER
***_reduce risk of Atherosclerosis_***
64
**What are S/Sx of High Serum Lipids?**
**And What CAUSES this?**
* *_XANTHOMAS_** = swelling full of lipids
* *_Enlarged LIVER / SPLEEN_**
caused by:
**high calorie diet** / *lack of exercise* / **cigarettes**
**T2DM** / **corticosteroids / estrogens / retinoids
B-adrenergic blocking agents**
**Optimal fasting serum levels:**
Total cholesterol \< 200 mg/dL
LDL cholesterol \< 100 mg/dL
HDL cholesterol \> 40 mg/dL
TG \< 150 mg/dL
65
What are **Routine Assay methods for**
## Footnote
**Cholesterol & Triglycerides?**
_Serum / Plasma Sample_ + enzymes
**Measure H2O2 = hydrogen peroxide --\> colored dye**
measured **_SPECTROPHOTOMETRICALLY_**
similar for both Cholesterol / TG's
66
What are Routine Assay methods for
## Footnote
**HDL?**
Precipitate HDL containing **_apo B-100_**
using cetrifugation and **heparin-MnCl2**
measure **_cholesterol REMAINING in the SUPERNATANT_**
| (VLDL + LDL + chylomicrons)
67
What are Routine Assay methods for
## Footnote
**LDL?**
Selectively **precipitate LDL**, using **polyvinyl sulfate or heparin**
@ *low pH,* then measure cholesterol in supernatent
## Footnote
**LDL = (Supernatent value) -MINUS- (TOTAL Cholesterol)**
68
What are Routine Assay methods for
## Footnote
**Total LipoProtein**
**_NMR_**
to measure lipoprotein associated FA methyl groups
**_Electrophoresis_**
to seperate lipoproteins
69
What does **Dehydration result in?**
And what are the **3 types of Dehydration?**
**_HYPER-Natremia_** + **_HYPER-Osmolarity_**
* *HYPER-Natremic** Dehydration
from: _water & food deprivation_ / sweating / diuresis / diuretic
* *Normo****Natremic** Dehydration
from: _vomiting & diarrhea_
* **HypoNatremic*** Dehydration:
from: _diuretic therapy / *inefficient adrenocorticoids*_ + renal disease
| (Na+ is high extracellularly, Na out)
70
What CAUSES **Overhydration?**
What is the RESULT?
*_NOT caused by Excessive Water Intake (POLYDIPSIA)_*
typically caused by **IMPAIRED RENAL FUNCTION**
too much water is retained by the kidney
***_hypoNatremia***_, _***hypo-osmality_***
71
What is **EXCESS Sodium** associated with?
Na Out
Too much dietary salt
**CHF** + **Liver / Renal Disease**
**Pregnancy**
**HYPER-Aldosteronism**
**_AAS / FES / Ion-Selective Electrode_**
72
What is **Sodium depletion associated with?**
**Vomitting / Diarrhea**
**Sweating**
* *Renal** / ***_hypo-aldosteronism_***
* *DM**
**BURNS**
_AAS / FES / Ion-Selective Electrode_
73
what is **HYPER-Kalemia** associated with?
* *K**idney is the major organ for **excretion**
* *K - IN**, very steep gradient **INSIDE**
Kidney disease
**_Crush / Crash Injuries_**
**Damaged Cells -\> LEAK POTASSIUM**
AAS / FES / Ion-Selective Electrode
74
**Chloride Balance**
CL intake is mainly through **Diet**
Excretin is mainly through **URINE**, *little through **FECES***
HYPER / Hypo - Chloremia
is associated with **SODIUM** excess or depletion
*exceptions can occur during **_acidosis / alkalosis_***
Color-agents + spectral changes / Ion-Selective Electrode
75
**Typical Methods for Assaying**
**ELECTROLYTES**
**_ION-SELECTIVE ELECTRODE_**
for **Sodium / Potassium / Chloride**
Sweat
76
**How do we Sample / TEST for this electrolyte?**
**Sodium**
Sample:
* *Serum / Plasma**
* *Urine / Feces / GI Fluids**
_**AAS** = Atomic Absorption Spectrophotometry_
_**FES** = Flame Emission Spectrophotometry_
**_Ion-Selective Electrode_**
*may also monitor a **enzyme or chromophore***
77
**How do we Sample / TEST for this electrolyte?**
**Potassium**
**K IN**
Sample:
**Serum / Plasma** ( ***_no RBCs that could release K+)_***
_**AAS** = Atomic Absorption Spectrophotometry_
_**FES** = Flame Emission Spectrophotometry_
**_Ion-Selective Electrode_**
78
**How do we Sample / TEST for this electrolyte?**
**Chloride**
Sample:
**Urine / Plasma / Serum**
Titration with **color agents** + monitor **spectral changes**
**_Ion-Selective Electrode_**
79
**How do we test for CYSTRIC FIBROSIS?**
**SWEAT**
Assay for:
**Conductivity**
**CHLORIDE SPECIFIC ELECTRODE**
ion-selective electrode
80
**LIVER PATHOLOGIES**
Problems with EXCRETION:
* *_Bile Acids**_ & _**Jaundice_**
* *_Xenobiotic Metabolis_****_m_** = DRUG Metabolism
**Hepatic _Synthetic Function Dysruption:_**
metabolism of Carbs / Fats / Protein
**Cirrhosis** / portal HYPER tention / **Varices / Ascites**
**Coagulation Factors**
**GALLSTONES**
81
**What is CHOLESTASIS?**
and how is it **Diagnosed?**
**_Bile is NOT flowing_** from the **liver -\> duodenom**
due to:
**gallstones / malignant growths
liver disease / drugs**
**_ALKALINE PHOSPHATASE = ALP_**
82
**What does Bilirubin / frationation Test diagnose?**
**Jaundice**
Disorders of **metabolism**
**Jaundice in the NEWBORN**
83
**What does AST / ALT test for?**
**Aspartate** / **Alanine** Aminotransferase
**AST =** Sensitive test of **Hepatocellular Disease**
**ALT** = Sensitive + **More Specific** test of **Hepatocellular Disease**
84
What does **Albumin** test help **Dianose?**
Indicator of **SEVERITY & CHRONICITY**
## Footnote
**of hepatic fxn / disease**
85
What does **Prothrombin Time test DIAGNOSE for?**
Test for liver disease's **SEVERITY**
**_Cholestasis_**
also is tested by **ALP**
86
What is the **biochemical origin of BILIRUBIN**?
How is it **Excreted?**
**_Heme Breakdown product_**
**Conjugated with GLUCURONIC ACID** --\> excreted in the **FECES**
= **Direct Bilirubin**
*Indirect bilirubin = unconjugated form*
**Urobilinogen** = *reduced form of bilirubin*
occurs when **liver's metablic capacity is exceeded**
detected by a **color change rxn in urine**
87
How do we **Assay BILIRUBIN**?
**_Direct Bilirubin Assay_**
react with **diazotized sulfanilic acid** w/ **serum / plasma**
measure **color change**
**_Urine Specimen_**:
**dipstick** **impregnated w/ diazo reagent**
also gives **color change**
88
**Assay difference between
Conjugated / Unconjugated / Total
BILIRUBIN**
**_Total Bilirubin_**
Seperate assay with **alcohol to ACCELERATE RXN**
**Conjugated = Direct Bilirubin Assay**
reacted with **diazotized sulfanilic acid**, measure color change
* **UNconjugated = Indirect Bilirubin***, ***_much slower RXN_***
* *Total - Direct = *Indirect***
89
**How do we assay for**
**Total Protein & Albumin?**
**_Dye-Binding_**:
FAST & CHEAP, but *less delicate*
**_Biuret_**
chemical rxn -\> **color change**, dependent on **AMIDE bonds**
**_Lowry_**
chemical rxn -\> color change, dependend on **Tyr + Trp Content**
***Direct photometric**
Measure **UV absorbance** @ 200-225nm
not preffered due to interfering substances*
90
**What do BIURET / LOWRY assays test for?**
**_TOTAL PROTEIN / ALBUMIN_**
Biuret = amide bonds, color change
Lowry = Tyr + Trp content, color change
91
What are **assays for SPECIFIC Liver Proteins?**
ex.
**Complement Factors / Clotting Factors**
**AST / ALT / LDH** enzymes
**_IMMUNOASSAYS_**
kits are cheap, and fast + specific
*some cross-reactivity w/ isozymes*
**_Electrophoretic Assays_** of SERUM samples
good for seperating / quantating **isozymes**
92
**What are specific PATHOLOGIES of**
**Gastric / Pancreatic / Intestinal Function?**
and what are
**typical markers?**
**Tumors** of **GI / Pancreas**
**Ulcers / Pancreatitis**
**Cystic Fibrosis**
*variety maldigestion/malabsorption disorder*
**Hormones / Enzymes / Unabsorbed food compotents**
markers
Typically assayed by **_IMMONOASSAYS_**
93
**What are selected HORMONES / ENZYMES** that are tested for
**Gastric / Pancreatic / Intestinal Function?**
And how are they
**ASSAYED?**
MOST HORMONES = **_Immunoassay_**
**Gastrin / VIP / GIP
Pepsin/pepsinogen**
**_Trypsin / Elastase / Chymotrypsin_**
synthetic substratess -\> color change + IMMUNOASSAY
* *_Amylase / Lipase_**
* *_coupled enzyme reactions_** -\> colored products -\> **spectrophotometrically** + IMMUNOASSAY
94
What are **KEY MARKERS** in
**Bone / Mineral Disorders**?
And **how are they caused?**
_**INORGANICS**
**Ca / Mg / Phosphate**_
_from THERAPY_
* *Diuretics** -\> *lower Mg2+*, RAISE Ca2+
* *Lithium** -\> RAISE BOTH Mg + Ca
* *Vit D / A** -\> RAISE Ca
```
**Hormones** = **vit D** + **ParaThyroid hormone**
Enzymes = **alkaline phosphatase**
```
95
How do we **Test / Assay** for this inorganic marker for **Bone/mineral disorders?**
**_Phosphate_**
Use **Serum** or **Heperinized Plasma**,
*EDTA/other anticoags interfere with assay*
ammonium molybdate -\>
**_UV-Vis Abosprtion_**
96
How do we **Test / Assay** for this inorganic marker for **Bone/mineral disorders?**
**_Calcium / Magnesium_**
Using **chelating dyes** -\> **colored complex**
## Footnote
**_UV-Vis Absorption_**
**_AAS_**
**_Ion-Selective Electrodes_**
97
How do we **Test / Assay** for this **hormone/enzyme** for **Bone/mineral disorders**?
## Footnote
**_Vitamin D & Metabolites_**
column chromatography
**HPLC** / **UV Absorption / Immunoassay**
98
How do we **Test / Assay** for this **hormone/enzyme** for **Bone/mineral disorders?**
**_IMMUNOASSAYS_**
for all
calcitonin / PTH / collagen telepeptides / osteocalcin
99
How do we **Test / Assay** for this **hormone/enzyme** for **Bone/mineral disorders?**
## Footnote
**_Alkaline Phosphates_**
marker for **BONE FORMATION**
**_Immunoassay_**
or **Complicated enzymatic Methods**
100
What are typical **markers for TUMORS?**
**and how are they tested for?**
May express **Aberrant Proteins** or **Excess "normal" proteins**
**Unusual carbs / polysaccherides
Mutated Genes**
**_IMMUNOASSAYS_**
some use **DNA sequence detection**
***mass spectrometry***
