Hi Laboni,

In addition to multiple molecular forms of cytochrome P450 in the liver, almost all beneath key liver tests might be assessed by colourmetric methods in blood samples. The panel usually consists of several tests that are run at the same time. These tests typically include:

Alanine aminotransferase (ALT) – an enzyme mainly found in the liver; the best test for detecting hepatitis

Alkaline phosphatase (ALP) – an enzyme related to the bile ducts but also produced by the bones, intestines, and duringpregnancy by the placenta (afterbirth); often increased when bile ducts are blocked.

Aspartate aminotransferase (AST) – an enzyme found in the liver and a few other organs, particularly the heart and other muscles in the body

Bilirubin – two different tests of bilirubin often used together (especially if a person has jaundice): total bilirubin measures all the bilirubin in the blood; direct bilirubin measures a form that is conjugated (combined with another compound) in the liver.

Albumin – measures the main protein made by the liver; the level can be affected by liver and kidney function and by decreased production or increased loss.

Total protein (TP) – measures albumin and all other proteins in blood, including antibodies made to help fight off infections

Other tests that may be included in a liver panel, such as:Gamma-glutamyl transferase (GGT) – another enzyme found mainly in liver cells

Lactate dehydrogenase (LD) – an enzyme released with cell damage; found in cells throughout the body

Prothrombin time (PT) – the liver produces proteins involved in the clotting (coagulation) of blood; the PT measures clotting function and, if abnormal, may indicate liver damage.

Alpha-feto protein (AFP) – associated with regeneration or proliferation of liver cell

Autoimmune antibodies (e.g., ANA, SMA, anti-LKM-1) – associated with autoimmune hepatitis.

Best wishes,

Ilya

Hi,

Dr. Tsyrlov wrote the important tests to know either liver is functioning well or not. Thanks Dr. Tsyrlov. But I would like to know what is the mechanism of the diagnostic kits used in the diagnosis of  ALP, ALT, ASP?

Best regards,

Didarul

I can only speak to the testing performed in my lab, but I believe these are very common method.  AST, ALT, and LDH all use NADH to NAD reduction reactions.  The others use various reactions. I can provide you with more details if this is the information you are looking for.

Best Luck

David  

Dear David,

Hi, Thanks for your answer. I am a student of Dr. Laboni and working under her supervision. Could you please give me more details about that, especially the principles, methods and detection techniques?

Best regards

Didarul Alam

Sayad,

Sorry for such a long post but it is a complex answer.  I hope this is what you need.

ALP reagent is used to measure alkaline phosphatase activity by a kinetic rate method using a 2-amino-2-methyl-1-propanol (AMP) buffer. In the reaction, alkaline phosphatase catalyzes the hydrolysis of the colorless organic phosphate ester substrate, p-nitrophenylphosphate, to the yellow colored product, p-nitrophenol, and phosphate. This reaction occurs at an alkaline pH of 10.3.  The ratio used is one part sample to 50 parts reagent. The system monitors the change in absorbance at 410 nanometers. This change in absorbance is directly proportional to the activity of ALP in the sample and is used by the System to calculate and express ALP activity.

The AST reagent is used to measure aspartate aminotransferase activity by an enzymatic rate method.1,2 In the assay reaction, the AST catalyzes the reversible transamination of L-aspartate and α-ketoglutarate to oxaloacetate and L-glutamate. The oxaloacetate is then reduced to malate in the presence of malate dehydrogenase (MDH) with the concurrent oxidation of β-Nicotinamide Adenine Dinucleotide (reduced form) (NADH) to β-Nicotinamide Adenine Dinucleotide (NAD). The ratio used is one part sample to 11 parts reagent. The system monitors the rate of change in absorbance at 340 nanometers over a fixed-time interval. This rate of change in absorbance is directly proportional to the activity of AST in the sample and is used by the SYNCHRON® System(s) to calculate and express the AST activity.

The AST reagent is used to measure aspartate aminotransferase activity by an enzymatic rate method.1,2 In the assay reaction, the AST catalyzes the reversible transamination of L-aspartate and α-ketoglutarate to oxaloacetate and L-glutamate. The oxaloacetate is then reduced to malate in the presence of malate dehydrogenase (MDH) with the concurrent oxidation of β-Nicotinamide Adenine Dinucleotide (reduced form) (NADH) to β-Nicotinamide Adenine Dinucleotide (NAD).  The ratio used is one part sample to 11 parts reagent. The system monitors the rate of change in absorbance at 340 nanometers over a fixed-time interval. This rate of change in absorbance is directly proportional to the activity of AST in the sample and is used by the SYNCHRON® System(s) to calculate and express the AST activity.

LD reagent is used to measure lactate dehydrogenase activity by an enzymatic rate method.2,3 In the reaction, LD catalyzes the reversible oxidation of L-lactate to pyruvate with the concurrent reduction of β-nicotinamide adenine dinucleotide (NAD) to reduced β-nicotinamide adenine dinucleotide (NADH). The ratio used is one part sample to 20 parts reagent. The system monitors the change in absorbance at 340 nanometers. This change in absorbance is directly proportional to the activity of lactate dehydrogenase in the sample and is used by the System to calculate and express the lactate dehydrogenase activity.

The AST reagent is used to measure aspartate aminotransferase activity by an enzymatic rate method.