Usage of Total Bile Acid Test Kit

The usage method of total bile acid (TBA) determination kit may vary depending on different kit brands and models, but generally speaking, its basic principles and steps are similar. The following is a general usage method based on common total bile acid assay kits:

1、 Preparation work

Preparation of reagent kit: Ensure that the reagent kit is within its expiration date and stored and transported according to the instructions. A reagent kit usually includes reagents (such as buffer solutions, enzyme-linked immunosorbent assay reagents, color reagents, etc.), standards, microplates, samplers, washing solutions, etc.

Sample preparation: Prepare the samples to be tested, such as serum, plasma, urine, etc., according to the instructions of the reagent kit. The sample should be fresh and free of contamination, and pre treated according to the instructions (such as centrifugation, dilution, etc.).

2、 Operation steps

Sample addition:

Set blank wells, standard wells, and test sample wells on the enzyme-linked immunosorbent assay (ELISA) coated plate.

According to the instructions, add the corresponding reagents and samples into the blank well, standard well, and test sample well, respectively. Be careful not to touch the hole wall when adding samples, gently shake and mix well.

Nurturing:

After sealing the plate with a sealing film, place the enzyme-linked immunosorbent assay (ELISA) plate in a 37 ℃ constant temperature incubator and incubate for a certain period of time (such as 30 minutes).

wash:

Be careful to remove the sealing film, discard the liquid, and shake dry.

Fill each well with detergent, let it sit for a certain period of time (such as 30 seconds), then discard it. Repeat this process multiple times (such as 5 times) and pat dry.

Enzyme addition:

Add a certain amount of enzyme-linked immunosorbent assay reagent to each well (excluding blank wells).

Re incubation:

Place the enzyme-linked immunosorbent assay (ELISA) plate again in a 37 ℃ constant temperature incubator and incubate for a certain period of time (such as 30 minutes).

wash:

Repeat the washing process of step 3.

Color rendering:

Add a certain amount of color developer A to each well first, then add color developer B, gently shake and mix well.

Color development for a certain period of time (such as 15 minutes) under light avoidance conditions at 37 ℃.

Termination:

Add a certain amount of termination solution to each well to terminate the reaction (at this point, the blue color turns yellow).

determination:

Measure the absorbance (OD value) of each well in sequence at a wavelength of 450nm using a blank air conditioner. The measurement should be conducted within 15 minutes after adding the termination solution.

3、 Result calculation

Draw a standard curve: Plot the standard curve on a coordinate paper with the concentration of the standard substance as the horizontal axis and the OD value as the vertical axis.

Calculate sample concentration: Based on the OD value of the sample, find the corresponding concentration on the standard curve. If a dilution factor is used, the detected concentration needs to be multiplied by the dilution factor to obtain the actual concentration of the sample.

4、 Precautions

Storage and use of reagent kits: The reagent kits should be stored in a dark environment at 2-8 ℃, avoiding direct sunlight and high temperatures. After opening, it should be used up within the specified time to avoid prolonged exposure to air.

Sample processing: The sample should be pre processed according to the instructions, such as centrifugation, dilution, etc. Avoid using expired or contaminated samples.

Accuracy of operation: When adding samples, a sampler should be used and its accuracy should be regularly checked. Avoid cross contamination and misoperation.

Interpretation of results: The measurement results should be compared with reference values and comprehensively judged based on clinical information. If there are any abnormal results, a re examination or further examination should be conducted.

Please note that the specific operating steps and precautions may vary depending on the brand and model of the reagent kit. Therefore, please read the instructions carefully before use and operate according to the requirements of the instructions.