Separation method and application of single coccidian oocysts

Abstract

The invention provides a separation method and application of single coccidian oocysts, and relates to the technical field of separation of coccidian oocysts. The separation method comprises the following steps: marking and positioning the bottom of a transparent container, dropwise adding oocyst liquid onto the gel, observing whether the dropping liquid on the surface of the gel contains a single oocyst or not through a microscope, recording the position containing the single oocyst, digging blank gel as a sealing top by using a suction tube, and separating the oocyst liquid from the sealing top. And then the gel containing the single oocyst dropping liquid is dug as a pad bottom, so that the single oocyst can be simply and conveniently fixed in the gel block. The separation method is simple and easy to operate, the separation time can be shortened, and the separation efficiency is improved.

Description

technical field [0001] The invention relates to the technical field of separation of coccidia oocysts, in particular to a separation method and application of a single coccidia oocyst. Background technique [0002] Chicken coccidia infection is self-limiting, and will be emptied in the host within a period of time, and the host specificity is very high; chicken coccidia are inoculated orally, and the inoculation method is simple and easy to implement. Based on the above advantages, chicken coccidia is an excellent vaccine carrier. Through gene editing technology, the antigen gene is knocked into the genome of chicken coccidia, making it express a constitutive antigen protein, so that the body can obtain immunity against the antigen. During this process, it is usually necessary to add a fluorescent tag protein to screen gene editing positive strains. Therefore, it is an essential step to isolate a single coccidian expressing a fluorescent tag to make it a pure gene editing p...

AI Technical Summary

Problems solved by technology

However, this method has the following defects: 1. The material preparation process is long and the operation is complicated. It takes a lot of time to cut the cellophane into small pieces with a length and width of 0.5 cm. During this period, it is necessary to ensure that the fragile cellophane is not broken and does not fly away. , it takes a lot of time; dripping the coccidia oocyst liquid on the cellophane, the cellophane is prone to accidental movement, causing the cellophane of the selected single oocyst to be overturned, causing the oocyst liquid to stick to the storage table or several pieces of paper overlap and stick together, etc. , can only be re-selected, which is a waste of time

2. Since the paper is not easy to fix and store, and the moving distance cannot be too far, animals must be inoculated at the place where single oocysts are separated, which may easily cause animals to be contaminated by other oocysts, and also limit the animal inoculation site and the isolation laboratory of single oocysts the distance

3. Drop the oocysts on the paper, the liquid evaporates quickly, and the oocysts are quickly in a dry and unfavorable environment, which reduces the activity of the oocysts

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