Description:Title of the Invention
“A Diagnostic Test Kit for Total Protein”
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 Total Protein from blood serum / plasma in vitro for laboratories and medical professionals.
Background of Invention
Plasma proteins are synthesized primarily in the liver, but also in plasma cells, lymph nodes, the spleen and bone marrow. Disease processes will affect the overall concentration of protein, as well the ratio of the protein fractions that comprise the total amount. Hypoproteinemia can be caused by loss of blood, sprue, nephrotic syndrome, severe burns, salt retention syndrome and insufficient dietary protein intake (Kwashiorkor). Hyperproteinemia may be observed in acute dehydration and illnesses like multiple myeloma. Changes in the relative amounts of the protein fractions can occur without a change in the total protein. The albumin/globulin ratio is commonly used to screen for abnormal protein fraction concentrations. Marked changes in this ratio can be observed in liver cirrhosis, glomerulonephritis, nephritic syndrome, acute hepatitis, lupus, and some chronic and acute inflammations.
This total protein method utilizes the biuret reaction, with measurement of the final product at 546 nm. Divalent copper reacts in alkaline solution with protein peptide bonds to form the characteristic purple-colored biuret complex. Sodium potassium tartrate prevents the precipitation of copper hydroxide and potassium iodide prevents auto reduction of copper. The color intensity is directly proportional to the protein concentration.
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 Total Protein.
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 Total Protein 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 Total Protein from blood serum / plasma useful effectively.
The secondary objective of the invention is that the diagnostic test kit for Total Protein utilizes photometric method herein the kit configured with suitable reagents like Sodium Hydroxide, Potassium Sodium Tartrate, Copper Sulphate and Total Protein as standard taken in effective concentration with 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 Total Protein is determined through calculation by the absorbance of sample and standard.
Summary of the Invention

The present disclosure relates a diagnostic kit specifically to determine Total Protein 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 Total Protein concentration.
A diagnostic test kit for Total Protein comprising a test kit characterized in that 100 ml size kit of reagent preferably consist of Sodium Hydroxide, Potassium Sodium Tartrate and Copper Sulphate with Total Protein as standard, estimates Total Protein in vitro from blood serum / plasma sample effectively, wherein reagent cell composed of 0.1 N of Sodium Hydroxide, 16 mmol/l of Potassium Sodium Tartrate, and 6 mmol/l of Copper Sulphate, taken with blood serum/ plasma sample from 0.020 ml to 0.025 ml into two clean test tube for test sample and blank sample and Total Protein 6 gm/dl sample as standard in third test tube; pipette out the reagent from the kit separately; transferring said reagent into the test tubes respectively at room temperature (37oC) and , uniformly mixed with test sample, blank sample and standard; incubating the reaction mixture for 15 minutes; determining the absorbance of said test sample blank sample and standard Total Protein sample on photometer at a wavelength of 546 nm for 30-90 seconds; and finally calculating the absorbance to estimate Total Protein from the test serum / plasma sample effectively.

One of the preferred embodiments of the present invention is the unique design of the kit prefilled with the preferred reagent like Sodium Hydroxide, Potassium Sodium Tartrate, Copper Sulphate reacts with serum/ plasma Total Protein evaluated through the absorbance on photometer.

One of the preferred embodiments of the present invention is that to determine Total Protein from the varied concentration more preferably making the test sample, blank sample and standard Total Protein sample to infer the clinical findings effectively.

One of the preferred embodiments of Total Protein 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 Total Protein

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
Plasma proteins are synthesized primarily in the liver, but also in plasma cells, lymph nodes, the spleen and bone marrow. Disease processes will affect the overall concentration of protein, as well the ratio of the protein fractions that comprise the total amount. Hypoproteinemia can be caused by loss of blood, sprue, nephrotic syndrome, severe burns, salt retention syndrome and insufficient dietary protein intake (Kwashiorkor). Hyperproteinemia may be observed in acute dehydration and illnesses like multiple myeloma. Changes in the relative amounts of the protein fractions can occur without a change in the total protein.
A diagnostic test kit for Total Protein comprising a test kit characterized in that 100 ml size kit of reagent preferably consist of Sodium Hydroxide, Potassium Sodium Tartrate and Copper Sulphate with Total Protein as standard, estimates Total Protein in vitro from blood serum / plasma sample effectively, wherein reagent cell composed of 0.1 N of Sodium Hydroxide, 16 mmol/l of Potassium Sodium Tartrate, and 6 mmol/l of Copper Sulphate, taken with blood serum/ plasma sample from 0.020 ml to 0.025 ml into two clean test tube for test sample and blank sample and Total Protein 6 gm/dl sample as standard in third test tube; pipette out the reagent from the kit separately; transferring said reagent into the test tubes respectively at room temperature (37oC) and , uniformly mixed with test sample, blank sample and standard; incubating the reaction mixture for 15 minutes; determining the absorbance of said test sample blank sample and standard Total Protein sample on photometer at a wavelength of 546 nm for 30-90 seconds; and finally calculating the absorbance to estimate Total Protein from the test serum / plasma sample effectively.
The present disclosure relates a diagnostic kit specifically to determine Total Protein concentration in the blood serum/plasma wherein the desired size kit is developed, essentially consists of reagents for reaction with blood serum/plasma samples, detected on photometer at desired wavelength to measure the absorbance and finally calculated the Total Protein concentration.
Preferred embodiments of a diagnostic kit for Total Protein
i) Diagnostic Kit: The Total Protein kit comprises of 100 ml size wherein reagent cell prefilled for the determination of Uric Acid, herein the key reagent like Sodium Hydroxide, Potassium Sodium Tartrate, and Copper Sulphate reacts with serum/ plasma Total Protein used in different concentration, estimates serum / plasma Total Protein through the absorbance using photometer of desired wavelength.
ii) Reagents: Blood serum /plasma, Sodium Hydroxide, Potassium Sodium Tartrate and Copper Sulphate.
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 0.020 ml to 0.025 ml
Reagent 0.1 N of Sodium Hydroxide, 16 mmol/l of Potassium Sodium Tartrate, and 6 mmol/l of Copper Sulphate
Shelf Life 24 months
Kit Size 100 ml
Wavelength 546 nm
Temperature 37oC
Time of Estimation 30 to 90 seconds

Table no. 02: Method to operate Total Protein

Particulars Blank Standard Sample
Reagent 1.0 ml 1.0 ml 1.0 ml
Standard -- 0.020 ml --
Serum/plasma test sample -- -- 0.020 to 0.025 ml

General Method to operate the device

Taking the 0.020 to 0.025 ml blood serum/ plasma sample as test sample, reagent as blank sample and Total Protein as standard separately in clean test tubes;
pipette out the reagent 0.1 N of Sodium Hydroxide, 16 mmol/l of Potassium Sodium Tartrate, and 6 mmol/l of Copper Sulphate from the kit separately; transferring said reagent into the test tubes respectively at room temperature (37oC); incubating the reaction mixture for 5 minutes; determining the absorbance of said test sample, blank sample and Total Protein 6 gm/dl sample as standard on photometer at a wavelength of 546 nm for 30-90 seconds; and finally calculating the absorbance to estimate Total Protein from the test serum / plasma sample effectively.

The absorbance is calculated using below formula:
Concentration = Absorption of sample / Absorption of standard x 6 gm/dl
(Table No. 1, 2 and Figure No. 1)
Example 1
Taking the 0.020 ml blood serum/ plasma sample as test sample, reagent as blank sample and Total Protein as standard separately in clean test tubes;
pipette out the reagent 0.1 N of Sodium Hydroxide, 16 mmol/l of Potassium Sodium Tartrate, and 6 mmol/l of Copper Sulphate from the kit separately; transferring said reagent into the test tubes respectively at room temperature (37oC); incubating the reaction mixture for 5 minutes; determining the absorbance of said test sample, blank sample and Total Protein 6 gm/dl sample as standard on photometer at a wavelength of 510 nm for 30-90 seconds; and finally calculating the absorbance to estimate Total Protein from the test serum / plasma sample effectively.

The absorbance is calculated using below formula:
Concentration = Absorption of sample / Absorption of standard x 6 gm/dl
Example 2
Taking the 0.025 ml blood serum/ plasma sample as test sample, reagent as blank sample and Total Protein as standard separately in clean test tubes;
pipette out the reagent 0.1 N of Sodium Hydroxide, 16 mmol/l of Potassium Sodium Tartrate, and 6 mmol/l of Copper Sulphate from the kit separately; transferring said reagent into the test tubes respectively at room temperature (37oC); incubating the reaction mixture for 5 minutes; determining the absorbance of said test sample, blank sample and Total Protein 6 gm/dl sample as standard on photometer at a wavelength of 510 nm for 30-90 seconds; and finally calculating the absorbance to estimate Total Protein from the test serum / plasma sample effectively.

The absorbance is calculated using below formula:
Concentration = Absorption of sample / Absorption of standard x 6 gm/dl
Scope of the Invention

A diagnostic test kit for Total Protein principally design to develop a simple method to analyse the Total Protein level using photometric method having size of 100 ml size kit of reagent preferably consist of effective concentration of 0.1 N of Sodium Hydroxide, 16 mmol/l of Potassium Sodium Tartrate, and 6 mmol/l of Copper Sulphate, taken with blood serum/ plasma sample from 0.020 ml to 0.025 ml as test sample and blank sample and Total Protein 6 gm/dl sample as standard respectively determines Total Protein 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 Total Protein comprising a test kit characterized in that 100 ml size kit of reagent preferably consist of effective concentration of 0.1 N of Sodium Hydroxide, 16 mmol/l of Potassium Sodium Tartrate, and 6 mmol/l of Copper Sulphate estimates Total Protein in vitro from blood serum / plasma sample taken from 0.020 to 0.025 ml as test sample and Total Protein ( 6 gm/dl) as standard, using photometer effectively.

2- The diagnostic test kit for Total Protein as claimed in claim 1, operates through in vitro diagnostic method to determine Total Protein comprises the following steps:
taking the 0.020 to 0.025 ml blood serum/ plasma sample as test sample, reagent as blank sample and Total Protein as standard separately in clean test tubes;
pipette out the reagent Sodium Hydroxide, Potassium Sodium Tartrate, and Copper Sulphate from the kit separately;
transferring said reagent into the test tubes respectively at room temperature (37oC);
incubating the reaction mixture for 5 minutes;
determining the absorbance of said test sample, blank sample and standard Total Protein sample on photometer at a wavelength of 546 nm for 30-90 seconds; and
calculating the absorbance to estimate Total Protein from the test serum / plasma sample effectively.