Milk antibody spectrum diagnostic kit and preparation method thereof

A diagnostic kit and antibody spectrum technology, applied in the field of enzyme immunoblotting, can solve problems such as false positives, uncertain differences in allergic protein allergies, false negatives, etc.

Inactive Publication Date: 2012-08-01
TIANJIN NANKAI HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The present invention aims to solve the problem that the current milk-specific diagnostic kit can only screen out the presence of milk-specific IgE in serum, but cannot determine the difference in patients' sensitivity to different allergenic proteins in milk, and the d

Method used

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  • Milk antibody spectrum diagnostic kit and preparation method thereof
  • Milk antibody spectrum diagnostic kit and preparation method thereof
  • Milk antibody spectrum diagnostic kit and preparation method thereof

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Embodiment Construction

[0054] The present invention will be described in detail below in conjunction with the accompanying drawings and embodiments.

[0055] 1. Solution preparation

[0056] ⑴ Phosphate buffered saline (PB) 0.1M pH 7.4

[0057] Double distilled water (ddH 2 O) 3.6 L

[0058] Sodium dihydrogen phosphate (NaH 2 PO 4 2H 2 O) 11.84 g

[0059] Disodium hydrogen phosphate (Na 2 HPO 4 12H 2 O) 116 g

[0060] Magnetic stirring, after fully dissolved, adjust the pH to 7.4, add ddH 2 O is fixed to 4L.

[0061] (2) Nitrocellulose membrane treatment solution (hereinafter referred to as membrane treatment solution or TBS)

[0062] Double distilled water (dd H 2 O) 800ml

[0063] Tris (Tris) 6.055 g

[0064] Sodium azide (NaN 3 ) 1g

[0065] Sodium chloride (NaCl) 8.766 g

[0066] Use HCl or NaOH to adjust the pH to 7.95-8.05, add dd H 2 O was adjusted to 1L.

[0067] ⑶ sample diluent

[0068] Double distilled water (ddH 2 O) 400ml

[0069] Tris (Tris) 3.03 g

[0070] Sodi...

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Abstract

The invention discloses a milk antibody spectrum diagnostic kit and a preparation method thereof, and belongs to an enzyme-linked immunoblotting technology. The kit comprises a chromogenic substrate, a blotting reaction tank, a mouse anti-human immunoglobulin E-alkaline phosphatase (IgE-AP) labeled antibody, sample diluent, enzyme-labeled antibody diluent, wash concentrate, a detection zone which is coated with casein and marked as T1, a detection zone which is coated with macromolecular protein and marked as T2, a detection zone which is coated with alpha-lactoalbumin and marked as T3, a detection zone which is coated with beta-lactoglobulin and marked as T4, and a quality control region which is coated with a rabbit anti-mouse immunoglobulin G (IgG) antibody and marked as C. Four allergens are coated in different zones at the same concentration, so that the same effect (specificity and sensitivity) can be ensured when IgE with different specificity is detected; and meanwhile, the detection result can indicate that specific antibodies of a plurality of allergen components exist in serum to be detected, clinical diagnosis accuracy can be improved, and the kit is suitable for further confirming milk allergy and making reasonable deallergization schemes.

Description

technical field [0001] The invention relates to enzyme immunoblotting technology, in particular to a milk antibody spectrum diagnostic kit and a preparation method thereof. Background technique [0002] Early detection of food allergy mainly focused on medical history observation and in vivo tests (such as skin prick test and food challenge test); however, these methods are not only expensive, time-consuming, but also have the risk of inducing severe allergic reactions, and have gradually been adopted. disuse. The essence of an allergic reaction is a wrong immune response. When the body is allergic to a certain food, it will produce specific antibodies (IgE and IgG) in the body, but such specific antibodies do not exist in the normal body. Therefore, at present, the detection of specific antibodies is often used clinically to determine the food allergy of patients, and finally to determine the allergic food. [0003] Using the method of in vitro detection of specific antib...

Claims

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Application Information

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IPC IPC(8): G01N33/68
Inventor 常艳敏李会强吴瑞杰李婵孙静文张爱民
Owner TIANJIN NANKAI HOSPITAL
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