Duck embryo early sex identification method and special kit

A kit and duck embryo technology, applied in measuring devices, instruments, scientific instruments, etc., to achieve the effects of low detection cost, stable source and good repeatability

Active Publication Date: 2015-06-10
武汉赛维尔生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Although the ELISA detection method and kit are a commonly used technology, so far there have been no reports on the use of indirect ELISA methods in poultry sex identification methods, especially in duck embryo sex identification methods.

Method used

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  • Duck embryo early sex identification method and special kit
  • Duck embryo early sex identification method and special kit
  • Duck embryo early sex identification method and special kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Example 1 Collection of Early Stage Samples and Preparation of Core Reagents

[0050] 1. Collection of duck embryo allantoic fluid

[0051] Take out the duck embryos from the incubator (the species are Jingjiang Shelduck or White Gai Duck, but not limited to these two species), put the big head up, let it stand for 3-5 minutes, use the egg canister to illuminate the eggs, and look for the location where the allantoic fluid is extracted At the position where the allantoic fluid is extracted from the positioning opening, avoid the distribution of blood vessels, mark it, disinfect the opening with 75% alcohol, pierce the eggshell with a disposable needle, and then extend it with a disposable sterile syringe or pipette. Insert the eggshell to extract the allantoic fluid, inject it into a sterile EP tube to obtain the allantoic fluid sample, and store it in a -20°C refrigerator for later use;

[0052] 2. Use the antigenic protein (estradiol protein) to coat the microtiter p...

Embodiment 2

[0067] The determination of embodiment 2 estradiol standard substance dilution concentration and the making of standard curve

[0068] The present invention determines that the concentration of the standard estradiol is successively set to 0ng / ml, 0.05ng / ml, 0.15ng / ml, 0.45ng / ml, 1.0ng / ml, and 2.0ng / ml after repeatedly exploring the measurement conditions.

[0069] Use CurveExpert software to fit the standard curve and obtain the standard curve formula.

[0070] Follow the steps below for testing:

[0071] a. Move the prepared reagent and the collected sample to be tested (ie duck embryo allantoic fluid) to room temperature of 20-25°C for at least 30 minutes to equilibrate;

[0072] b. Take a microplate, add the concentration gradient of the standard substance in each well of the microplate, then add 50 μ L of duck embryo allantoic fluid to be tested in order in each well, and then add estradiol monoclonal antibody (purchased 50 μL from Wuhan Huamei Bioengineering Co., Ltd.,...

Embodiment 3

[0080] Application example 1 of embodiment 3 detection method

[0081] The estradiol content in the allantoic fluid of white-modified duck embryos was determined by using the conventional allantoic fluid samples of white-modified duck embryos and the established standard curve formula, so that the sex of male and female embryos of white-modified ducks can be accurately identified. The specific method is as follows:

[0082] (1) Get 100ul of embryonic allantoic fluid of 14d embryonic ducks (collected from a duck farm in Fujian Province, China, but the present invention is not limited to this species), a total of 22 samples, judged with the naked eye according to the difference in the developmental shape of the male and female gonads. Record gender, including 10 male samples and 12 female samples, and store them at -20°C for later use;

[0083] (2) Determination of estradiol content by indirect competitive ELISA method [1] (operating steps according to Example 2). To establish...

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Abstract

The invention belongs to the technical field of poultry sex detection, and in particular relates to a duck embryo early sex identification method and a special kit. The duck embryo early sex identification method is established by collecting a hatched early duck embryo allantoic fluid, making use of a monoclonal antibody for estradiol detection and carrying out antigen-antibody reaction. Male and female sexes are judged according to a characteristic that the content of estradiol in a female embryo is higher than 1ng/ml and the content of the estradiol in a male embryo is lower than 0.6ng/ml. The identification method disclosed by the invention can identify the female and male sexes in an early hatching period without an influence on the further development of the embryo; and the identification method is conducive to animal welfare and relieving the transmission of vertical diseases.

Description

technical field [0001] The invention belongs to the technical field of sex identification of poultry, and in particular relates to a method for early sex identification of duck embryos and a special kit. Background technique [0002] The gender identification methods of ducks are generally appearance identification, anus touch identification, sound identification and molecular identification. The appearance identification method is based on the difference in body shape and appearance of male and female ducks, and often leads to misjudgments; although the anus identification method has a high accuracy rate, there is a risk of cross-infection of some vertically transmitted germs, and the anus touch has relatively high technical requirements and requires practical experience in production Only richness can ensure the accuracy; the sound identification method needs the duck's syrinx to be well developed, and it is difficult to distinguish between hatching and adolescence; althou...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577
CPCG01N33/543
Inventor 杨宇叶胜强龚萍钱运国杨永平王丽霞邓兵赵胜兰夏瑜
Owner 武汉赛维尔生物科技有限公司
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