Description:Title of the Invention
“A Diagnostic Test Kit for Serum Glutamic Pyruvic Transaminase (SGPT) / Alanine Aminotransferase (ALT)

Field of the Invention
The present disclosure relates generally to field of medical devices. More specifically the present invention relates to diagnostic test kit works on photometric method to determine A Diagnostic Test Kit for Serum Glutamic Pyruvic Transaminase (SGPT) / Alanine Aminotransferase (ALT) from blood serum / plasma in vitro for laboratories and medical professionals.
Background of Invention
The liver has a significant role in metabolism, digestion, detoxification, and elimination of substances from the body. The liver function tests typically include alanine transaminase (ALT) and aspartate transaminase (AST), alkaline phosphatase (ALP), gamma-glutamyl transferase (GGT), serum bilirubin, prothrombin time (PT), the international normalized ratio (INR), total protein and albumin. These tests can help determine an area of the liver where damage may be taking place and, depending on the pattern of elevation, can help organize a differential diagnosis. Elevations in ALT and AST disproportion to elevations in alkaline phosphatase and bilirubin denote hepatocellular disease. An elevation in alkaline phosphatase and bilirubin in disproportion to ALT and AST would characterize a cholestatic pattern. The actual function of the liver can be graded based on its ability to produce albumin as well as vitamin K-dependent clotting factors.
Elevated liver function tests are found in approximately 8% of the general population. These elevations may be transient in patients without symptoms, with up to 30% of elevations resolving after three weeks. Thus, care should be taken when interpreting these results to avoid unnecessary testing. A borderline AST and/or ALT elevation is defined as less than 2 times the upper limit of normal (ULN), a mild AST and/or ALT elevation as 2 to 5 times ULN, moderate AST and/or ALT elevation 5 to 15 times ULN, severe AST and/or ALT elevation greater than 15 times ULN, and massive AST and/or ALT greater than 10,000 IU/l.The magnitude of AST and ALT elevation varies depending on the cause of hepatocellular injury.
Typically when reviewing liver function tests, the discussion includes alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), gamma-glutamyl transferase (GGT), 5'nucleotidase, total bilirubin, conjugated (direct) bilirubin, unconjugated (indirect)bilirubin, prothrombin time (PT), the international normalized ratio (INR), lactate dehydrogenase, total protein, globulins, and albumin. These tests can help determine the area of hepatic injury, and the elevation pattern can help organize a differential diagnosis.

Therefore, there is a need for a device that overcomes at least the above mentioned problems. There is a need for a rapid, precise, and cost-effective diagnostic kit and method for the measurement of Serum Glutamic Pyruvic Transaminase (SGPT) / Alanine Aminotransferase (ALT) from blood serum / plasma in vitro for laboratories and medical professionals.
The present disclosure relates generally to field of medical devices. More specifically the present invention relates to diagnostic test kit works on photometric method to estimate Serum Glutamic Pyruvic Transaminase (SGPT) / Alanine Aminotransferase (ALT) from serum / plasma, in vitro for laboratories and medical professionals.
Object of the Invention
The primary object of the invention is to develop a diagnostic test kit to estimate Serum Glutamic Pyruvic Transaminase (SGPT) / Alanine Aminotransferase (ALT) from blood serum / plasma useful effectively.
The secondary objective of the invention is that the diagnostic test kit for Serum Glutamic Pyruvic Transaminase (SGPT) / Alanine Aminotransferase (ALT) utilizes photometric method herein the kit configured with suitable reagents like Tris, Laspartate, Lactate Dehydrogenase (LDH) and Nicotinamide Adenine Dinucleotide (NADH) reagent taken in effective concentration of blood serum /plasma.
Another objective of the invention is that the prepared blood serum/plasma samples with reagents estimated for absorbance in photometer at desired wavelength and room temperature.
Further objective of the invention is that Serum Glutamic Pyruvic Transaminase (SGPT) / Alanine Aminotransferase (ALT) is determined through calculation by multiplying the absorbance with factor.
Summary of the Invention

The present disclosure relates a diagnostic kit specifically to determine the Serum Glutamic Pyruvic Transaminase (SGPT) / Alanine Aminotransferase (ALT) concentration in the blood serum/plasma wherein the desired size kit is developed, essentially consist of reagents for reaction with blood serum/plasma samples, detected on photometer at desired wavelength to measure the absorbance and finally calculated to determine the Serum Glutamic Pyruvic Transaminase (SGPT) / Alanine Aminotransferase (ALT) concentration.
A diagnostic test kit for Serum Glutamic Pyruvic Transaminase (SGPT) / Alanine Aminotransferase (ALT) comprising a test kit characterized in that 75 ml size kit of two compartment cell reagents preferably consist of Tris, Laspartate and Lactate Dehydrogenase (LDH) and Nicotinamide Adenine Dinucleotide (NADH) reagent determines Serum Glutamic Pyruvic Transaminase (SGPT) / Alanine Aminotransferase (ALT) in vitro from blood serum / plasma sample effectively, wherein the two cell reagent composed of reagent I cell of 80 mmol/l of Tris, 240 mmol/l of Laspartate and 600 µ/l of Lactate Dehydrogenase (LDH) and reagent II cell of 0.18 mmol/l of Nicotinamide Adenine Dinucleotide (NADH) determines Serum Glutamic Pyruvic Transaminase (SGPT) / Alanine Aminotransferase (ALT) by taking blood serum/ plasma sample from 0.080 ml to 0.100 ml into two clean test tube for test and blank sample; pipette out the reagent I and II from the kit separately in effective ratio of 4:1 respectively to determine Serum Glutamic Oxaloacetic Transaminase (SGOT) / Aspartate Aminotransferase (AST) ; transferring it into the test tubes respectively at room temperature (37oC); incubating the reaction mixture for 5 minutes; determining the absorbance of said test sample and blank sample on photometer at a wavelength of 340 nm for 60-180 seconds; and finally calculating the absorbance using factor to determine Serum Glutamic Pyruvic Transaminase (SGPT) / Alanine Aminotransferase (ALT) from the test serum / plasma sample effectively.

One of the preferred embodiments of the present invention is the unique design of the kit consist of two reagent cell (I & II ), which prefilled with the preferred reagent like Tris, Laspartate, Lactate Dehydrogenase (LDH), Malate Dehydrogenase (MDH) and Nicotinamide Adenine Dinucleotide (NADH) reacts with serum/ plasma Serum Glutamic Pyruvic Transaminase (SGPT) / Alanine Aminotransferase (ALT) evaluated through the absorbance on photometer.

One of the preferred embodiments of the present invention is that to determine Serum Glutamic Pyruvic Transaminase (SGPT) / Alanine Aminotransferase (ALT) from the varied concentration more preferably making the test sample and blank sample to infer the clinical findings effectively.

One of the preferred embodiments of that Serum Glutamic Pyruvic Transaminase (SGPT) / Alanine Aminotransferase (ALT) is estimated in vitro through all in one reagent kit.

Brief Description of Drawings

The following thorough explanation of the various aspects of the invention, taken in conjunction with the corresponding drawing that represents various aspects and other features of the disclosure invention.

Figure 1: Flow chart of method of estimation of Serum Glutamic Pyruvic Transaminase (SGPT) / Alanine Aminotransferase (ALT)

Detailed Description of the Invention

The following description is of exemplary embodiments only and is not intended to limit the scope, applicability or configuration of the invention in any way. Rather, the following description provides a convenient illustration through explanation and figures for implementing exemplary embodiments of the invention. Various changes to the described embodiments may be made in the function and arrangement of the ingredients described without departing from the scope of the invention.
The use of “including”, “comprising” or “having” variations thereof herein is meant to encompass the items listed thereafter and equivalents thereof as well as additional items. The terms “a” and “an” herein do not denote a limitation of quantity, but rather denote the presence of at least one of the referenced items. Further, the use of terms “first”, “second”, and “third”, and the like, herein do not denote any order, quantity, or importance, but rather are used to distinguish one element from another.
The terms used in the invention
mmol: milimole
mmol/l: millimole per liter
The liver has a significant role in metabolism, digestion, detoxification, and elimination of substances from the body. The liver function tests typically include alanine transaminase (ALT) and aspartate transaminase (AST), alkaline phosphatase (ALP), gamma-glutamyl transferase (GGT), serum bilirubin, prothrombin time (PT), the international normalized ratio (INR), total protein and albumin. These tests can help determine an area of the liver where damage may be taking place and, depending on the pattern of elevation, can help organize a differential diagnosis. Elevations in ALT and AST disproportion to elevations in alkaline phosphatase and bilirubin denote hepatocellular disease. An elevation in alkaline phosphatase and bilirubin in disproportion to ALT and AST would characterize a cholestatic pattern. The actual function of the liver can be graded based on its ability to produce albumin as well as vitamin K-dependent clotting factors.
A diagnostic test kit for Serum Glutamic Pyruvic Transaminase (SGPT) / Alanine Aminotransferase (ALT) comprising a test kit characterized in that 75 ml size kit of two compartment cell reagents preferably consist of Tris, Laspartate and Lactate Dehydrogenase (LDH) and Nicotinamide Adenine Dinucleotide (NADH) reagent determines Serum Glutamic Pyruvic Transaminase (SGPT) / Alanine Aminotransferase (ALT) in vitro from blood serum / plasma sample effectively, wherein the two cell reagent composed of reagent I cell of 80 mmol/l of Tris, 240 mmol/l of Laspartate and 600 µ/l of Lactate Dehydrogenase (LDH) and reagent II cell of 0.18 mmol/l of Nicotinamide Adenine Dinucleotide (NADH) determines Serum Glutamic Pyruvic Transaminase (SGPT) / Alanine Aminotransferase (ALT) by taking blood serum/ plasma sample from 0.080 ml to 0.100 ml into two clean test tube for test and blank sample; pipette out the reagent I and II from the kit separately in effective ratio of 4:1 respectively to determine Serum Glutamic Oxaloacetic Transaminase (SGOT) / Aspartate Aminotransferase (AST) ; transferring it into the test tubes respectively at room temperature (37oC); incubating the reaction mixture for 5 minutes; determining the absorbance of said test sample and blank sample on photometer at a wavelength of 340 nm for 60-180 seconds; and finally calculating the absorbance using factor to determine Serum Glutamic Pyruvic Transaminase (SGPT) / Alanine Aminotransferase (ALT) from the test serum / plasma sample effectively.

The present disclosure relates a dognostic kit specifically to determine Serum Glutamic Pyruvic Transaminase (SGPT) / Alanine Aminotransferase (ALT) concentration in the blood serum/plasma wherein the desired size kit is developed, essentially consist of reagents for reaction with blood serum/plasma samples, detected on photometer at desired wavelength to measure the absorbance and finally calculated to determine Serum Glutamic Pyruvic Transaminase (SGPT) / Alanine Aminotransferase (ALT) concentration.
Preferred embodiments of Serum Glutamic Pyruvic Transaminase (SGPT) / Alanine Aminotransferase (ALT)
i) Diagnostic Kit : The Serum Glutamic Pyruvic Transaminase (SGPT) / Alanine Aminotransferase (ALT) kit comprises of 75 ml size wherein two compartment cell consist prefilled reagent for the determination of blood serum/ plasma Serum Glutamic Pyruvic Transaminase (SGPT) / Alanine Aminotransferase (ALT), herein the key reagent Tris, Laspartate and Lactate Dehydrogenase (LDH) and Nicotinamide Adenine Dinucleotide (NADH) used in different concentration, effectively reacts with serum / plasma Serum Glutamic Pyruvic Transaminase (SGPT) / Alanine Aminotransferase (ALT) measures the absorbance using photometer of desired wavelength.
ii) Reagents: Blood serum /plasma, Tris, Laspartate, Lactate Dehydrogenase (LDH) and Nicotinamide Adenine Dinucleotide (NADH).
iii) Apparatus / Instrument: Test tubes, Photometer, Incubator etc.
Table no. 01: Preferred embodiments and parameters of the kit with concentrations

Particulars Name of ingredient with concentration
Serum/ plasma sample 0.080 ml to 0.100 ml
Reagent I 80 mmol/l of Tris, 240 mmol/l of Laspartate, and 600 µ/l of Lactate Dehydrogenase (LDH)
Reagent II 0.18 mmol/l of Nicotinamide Adenine Dinucleotide (NADH)
Effective Ratio of reagent I : reagent II 4:1
Shelf Life 24 months
Kit Size 75 ml
Wavelength 340 nm
Temperature 37oC
Time of Estimation 60 to 180 seconds
Factor 1746

Table no. 02: Method to operate Serum Glutamic Pyruvic Transaminase (SGPT) / Alanine Aminotransferase (ALT)

Particulars Blank Sample
Reagent I 4.0 ml 4.0 ml
Reagent II -- 1.0 ml
Serum/plasma test sample 0.080 to 0.100 ml 0.080 to 0.100 ml

General Method to operate the device
Taking the 0.080 to 0.100 ml blood serum/ plasma sample into two clean test tubes for test and blank sample separately;
pipette out the reagent I (80 mmol/l of Tris, 240 mmol/l of Laspartate and 600 µ/l of Lactate Dehydrogenase (LDH) and II (0.18 mmol/l of Nicotinamide Adenine Dinucleotide (NADH) in 4:1 ratio from the kit separately to prepare test sample and only reagent I (80 mmol/l of Tris, 240 mmol/l of Laspartate and 600 µ/l of Lactate Dehydrogenase (LDH) used to prepare blank sample; transferring it into the serum/ plasma sample respectively at room temperature (37oC) and uniformly mixed; incubating the reaction mixture for 5 minutes; determining the absorbance of said test sample and blank sample on photometer at a wavelength of 340 nm for 60-180 seconds; and finally calculating the absorbance using factor to Serum Glutamic Pyruvic Transaminase (SGPT) / Alanine Aminotransferase (ALT) from the test serum / plasma sample effectively.

The absorbance is calculated using below formula:
Serum Glutamic Pyruvic Transaminase (SGPT) / Alanine Aminotransferase (ALT) : 1746 x Absorbance
(Table No. 1, 2 and Figure No. 1)
Example 1
Taking the 0.080 ml blood serum/ plasma sample into two clean test tubes for test and blank sample separately;
pipette out the reagent I (80 mmol/l of Tris, 240 mmol/l of Laspartate, 600 µ/l of Lactate Dehydrogenase (LDH) and II (0.18 mmol/l of Nicotinamide Adenine Dinucleotide (NADH) in 4:1 ratio from the kit separately to prepare test sample and only reagent I (80 mmol/l of Tris, 240 mmol/l of Laspartate and 600 µ/l of Lactate Dehydrogenase (LDH) used to prepare blank sample; transferring it into the serum/ plasma sample respectively at room temperature (37oC) and uniformly mixed; incubating the reaction mixture for 5 minutes; determining the absorbance of said test sample and blank sample on photometer at a wavelength of 340 nm for 60-180 seconds; and finally calculating the absorbance using factor to Serum Glutamic Pyruvic Transaminase (SGPT) / Alanine Aminotransferase (ALT) from the test serum / plasma sample effectively.

The absorbance is calculated using below formula:
Serum Glutamic Pyruvic Transaminase (SGPT) / Alanine Aminotransferase (ALT) : 1746 x Absorbance
Example 2
Taking the 0.100 ml blood serum/ plasma sample into two clean test tubes for test and blank sample separately;
pipette out the reagent I (80 mmol/l of Tris, 240 mmol/l of Laspartate and 600 µ/l of Lactate Dehydrogenase (LDH) and II (0.18 mmol/l of Nicotinamide Adenine Dinucleotide (NADH) in 4:1 ratio from the kit separately to prepare test sample and only reagent I (80 mmol/l of Tris, 240 mmol/l of Laspartate and 600 µ/l of Lactate Dehydrogenase (LDH) used to prepare blank sample; transferring it into the serum/ plasma sample respectively at room temperature (37oC) and uniformly mixed; incubating the reaction mixture for 5 minutes; determining the absorbance of said test sample and blank sample on photometer at a wavelength of 340 nm for 60-180 seconds; and finally calculating the absorbance using factor to Serum Glutamic Pyruvic Transaminase (SGPT) / Alanine Aminotransferase (ALT) from the test serum / plasma sample effectively.

The absorbance is calculated using below formula:
Serum Glutamic Pyruvic Transaminase (SGPT) / Alanine Aminotransferase (ALT) : 1746 x Absorbance

Scope of the Invention

A diagnostic test kit for Serum Glutamic Pyruvic Transaminase (SGPT) / Alanine Aminotransferase (ALT) principally design to develop a simple method to analyse the serum / plasma Serum Glutamic Pyruvic Transaminase (SGPT) / Alanine Aminotransferase (ALT) level using photometric method having size of 75 ml size kit of two compartment cell reagent preferably consist of effective concentration of 80 mmol/l of Tris, 240 mmol/l of Laspartate and 600 µ/l of Lactate Dehydrogenase (LDH) and 0.18 mmol/l of Nicotinamide Adenine Dinucleotide (NADH) in effective ratio of 4:1 determines Serum Glutamic Pyruvic Transaminase (SGPT) / Alanine Aminotransferase (ALT) in vitro from blood serum / plasma sample effectively, could be potential diagnostic and economical method, compare to expensive conventional methods in future.

It is to be understood that the present invention is not limited to the embodiments described above, it should be clear that various modifications and alterations can be made along with various features of one embodiment included in other embodiments, within the scope of the present invention.
, Claims:We claim,
1- A diagnostic test kit for Serum Glutamic Pyruvic Transaminase (SGPT) / Alanine Aminotransferase (ALT) comprising a test kit characterized in that 75 ml size kit of two compartment cell reagent preferably consist of effective concentration of 80 mmol/l of Tris, 240 mmol/l of Laspartate and 600 µ/l of Lactate Dehydrogenase (LDH) (Reagent I) and 0.18 mmol/l of Nicotinamide Adenine Dinucleotide (NADH) (Reagent II) in effective ratio of 4:1 respectively, estimates Serum Glutamic Pyruvic Transaminase (SGPT) / Alanine Aminotransferase (ALT) in vitro from blood serum / plasma sample taken from 0.080 to 0.100 ml, using photometer effectively.

2- The diagnostic test kit for Serum Glutamic Pyruvic Transaminase (SGPT) / Alanine Aminotransferase (ALT) as claimed in claim 1, operates through in vitro diagnostic method to determine Serum Glutamic Pyruvic Transaminase (SGPT) / Alanine Aminotransferase (ALT) comprises the following steps:
taking the 0.080 to 0.100 ml blood serum/ plasma sample into two clean test tubes for test and blank sample separately;

pipette out the reagent I (80 mmol/l of Tris, 240 mmol/l of Laspartate and 600 µ/l of Lactate Dehydrogenase (LDH) and II (0.18 mmol/l of Nicotinamide Adenine Dinucleotide (NADH) in 4:1 ratio from the kit separately to prepare test sample and only reagent I (80 mmol/l of Tris, 240 mmol/l of Laspartate and 600 µ/l of Lactate Dehydrogenase (LDH) used to prepare blank sample;

transferring it into the serum/ plasma sample respectively at room temperature (37oC) and uniformly mixed;

incubating the reaction mixture for 5 minutes;

determining the absorbance of said test sample and blank sample on photometer at a wavelength of 340 nm for 60-180 seconds; and

calculating the absorbance using factor to Serum Glutamic Pyruvic Transaminase (SGPT) / Alanine Aminotransferase (ALT) from the test serum / plasma sample effectively.