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Progesterone colloidal gold immunochromatography test strip for monkey early pregnancy detection

An immunochromatographic test strip and early pregnancy technology, which is applied in measuring devices, analytical materials, instruments, etc., can solve the problems of not being able to detect the physiological state of golden monkeys and macaques, and achieve the effects of easy operation, high sensitivity, and strong specificity

Active Publication Date: 2016-03-23
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

And before the present invention is put into practice, the existing early pregnancy detection test paper of human medicine is once tested to detect the early pregnancy of golden monkey and rhesus monkey, and the urine of golden monkey and rhesus monkey has also been tested with existing bovine progesterone detection test strip. The fluid was tested, but it could not detect the physiological state of early pregnancy in golden monkeys and rhesus monkeys

Method used

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  • Progesterone colloidal gold immunochromatography test strip for monkey early pregnancy detection
  • Progesterone colloidal gold immunochromatography test strip for monkey early pregnancy detection
  • Progesterone colloidal gold immunochromatography test strip for monkey early pregnancy detection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1 Preparation and Purification of Monoclonal Antibody

[0033] 1. Preparation of hybridoma cells: Splenocytes from mice with better immune effects were taken for cell fusion, and the mice were boosted immunized three days before the fusion, mixed with myeloma cells and splenocytes, treated with PEG-2000 and placed 37°C, 5% CO 2 cultured in a constant temperature incubator. Afterwards, according to the growth of the cells, the cell culture supernatant was taken, and positive cell wells secreting progesterone antibodies were screened out by ELISA method. The screened positive cell wells were cloned by the limiting dilution method, and finally a hybridoma cell line was established that could secrete a monoclonal antibody capable of detecting progesterone. The applicant named the hybridoma cell line as hybridoma cell line LX20150930 was deposited in the China Type Culture Collection (CCTCC) of Wuhan University, Wuhan, China on September 30, 2015, and the deposit n...

Embodiment 2

[0035] Example 2 Preparation of gold-labeled antibody-related solutions

[0036] 1. Preparation of various solutions:

[0037] (1) Preparation of trisodium citrate solution: Accurately weigh 0.5697 g of trisodium citrate, dissolve it in ultrapure water, and set the volume to 50 mL. Store at 4°C for later use.

[0038] (2) Preparation of chloroauric acid solution: Dissolve 1 g of solid chloroauric acid in ultrapure water, and dilute to 100 mL. Store at 4°C for later use.

[0039] (3) Preparation of potassium carbonate solution: 0.01mol / L, accurately weigh 1.38gK 2 CO 3 , dissolved in ultrapure water, and adjusted to 100 mL. Store at 4°C for later use.

[0040] (4) Preparation of phosphate buffered saline (PBS): Accurately weigh 1.00 g of bovine serum albumin (BSA) and dissolve it in a small amount of 0.01M pH 7.2 PB solution, and dilute to 100 mL with double-distilled deionized water.

[0041] (5) Preparation of 10% bovine serum albumin (BSA) solution: Accurately weigh 1...

Embodiment 3

[0064] The preparation of embodiment 3 progesterone colloidal gold immunochromatography test strips

[0065] 1. Determination of coating antigen dilution and concentration

[0066] (1) Determination of the best diluent:

[0067] Preparation of various dilutions:

[0068] a. 0.01mol / L TBS solution, pH=7.6

[0069] b. 0.01mol / L TBS solution, pH=9.0

[0070] c. 0.01mol / L PBS solution, pH=7.6

[0071] d. 0.01mol / L carbonate buffer solution, pH=9.0

[0072] e. Double distilled water

[0073] The coating antigen (progesterone-OVA, purchased from Shanghai Yaochao Bioengineering Co., Ltd.) was diluted to 0.2mg / ml, 0.6mg / ml, 1.0mg / ml, 1.4mg / ml with the above five solutions, and then used The film spotter coats the antigen on the test strip, and after drying, add the gold-labeled antibody dropwise, observe the results, and determine the best diluent.

[0074] (2) Determination of coating antigen concentration

[0075] Dilute the antigen to 0.2mg / ml, 0.6mg / ml, 0.8mg / ml, 1.0mg / ml,...

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Abstract

The invention belongs to the technical field of animal immunization biochemical detection, and particularly relates to a progesterone colloidal gold immunochromatography test strip for monkey early pregnancy detection. The test strip comprises a PCV bottom plate, an absorption pad, a nitrocellulose membrane, water absorption filter paper, a gold mark pad and a sample pad, wherein the nitrocellulose membrane is provided with a detection line and a quality control line, and the absorption pad, the nitrocellulose membrane, the gold mark pad and the sample pad are sequentially pasted to the PCV bottom plate. The test strip is characterized in that the gold mark pad contains progesterone monoclonal antibodies; the gold mark pad containing the progesterone monoclonal antibodies and the water absorption filter paper are arranged at the two ends of the nitrocellulose membrane respectively in an overlapping mode, and meanwhile the gold mark pad containing the progesterone monoclonal antibodies and the sample pad are arranged in an overlapping mode; the monoclonal antibodies capable of detecting monkey early pregnancy are excreted by a hybridoma cell strain with the preservation number of CCTCC NO:C2015170. The test strip is high in sensitivity and easy and convenient to operate and has great significance for improving breeding monitoring of monkeys such as macaques and snub-nosed monkeys.

Description

technical field [0001] The invention belongs to the technical field of animal immunochemical detection, and in particular relates to a progesterone colloidal gold immunochromatographic test strip for monkey early pregnancy detection. Background technique [0002] Monkey is a general term. Many animals in primates are called monkeys by us. Their brains are developed and they are the highest group among non-human primates. Monkeys are generally single-born animals, and very few can reach two or three. Macaques are the most common species of monkeys. Their metabolic characteristics, physiological functions and immune metabolism are very similar to those of humans. The breeding season is generally from September to April of the following year. Golden monkeys are also seasonal breeding animals. The obvious mating peak is generally in September-December, and production is generally in March-May. Macaques, golden monkeys and other monkeys are all wild animals under national key pr...

Claims

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Application Information

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IPC IPC(8): G01N33/577
CPCG01N33/577
Inventor 李翔韩丽杨利国肖运才马振伐陈礼刘锡玲陈建国郭爱珍滑国华
Owner HUAZHONG AGRI UNIV
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