Method for rapidly obtaining large amount of spore shells of nosema bombycis

Abstract

The invention provides a method for rapidly obtaining large amount of spore shells of nosema bombycis. The method comprises the following steps of adding 10<9>-10<20> 400muL of nosema bombycis into a solution, mixing the nosema bombycis suspension with 212-300mu m of glass beads according to the proportion of 1mu L:1mg to obtain a mixture, putting the mixture into a micro burette mixer and oscillating for 8-12 hours, carrying out gradient density centrifugation on the spore suspension, and taking sediment. According to the method, the spore shell extracting and purification processes are simplified, the operation is simple and fast, the requirement on equipment is low, the repeatability is high, a large amount of spores can be rapidly broken in one day so as to obtain the spore shells, the yield of the spore shell can reach 70-90% or even 100%, and the acquisition of the spore shell is not affected by the weakened vitality of the spores.

Description

technical field [0001] The invention relates to the field of biological technology, and relates to the technical field of diagnosis and identification of the microsporidia silkworm. Background technique [0002] Microsporidia are a group of pathogens that obligately parasitize intracellularly in the form of spores, and have a wide range of hosts. Almost all animals, including protozoa and humans, can be infected with Microsporidia. Nosema bombycis is It was isolated and identified from diseased silkworms in 1857. It can infect silkworms and cause silkworm microparticle disease, which is a devastating disease in sericulture production. Therefore, the detection of silkworm microsporidia is very important in sericulture. In recent years, According to the different antigenicity of the interspecies sporozoite protein of microsporidia, some immunological detection methods have been gradually developed. The immunological detection method presents a certain reaction phenomenon th...

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Problems solved by technology

However, in the germination method, on the one hand, the germination process requires high spore vigor to ensure the germination rate, and sufficient spore shells can be obtained after purification, but during the storage of the spores, the viability of the spores drops sharply, resulting in a very low spore germination rate. Only about 10-20%, even no germination so that the spores cannot be obtained, and the spores after a germination treatment can no longer germinate, so the germination method has great non-repeatability in operation, resulting in time and cost. Waste; on the other hand, because the germination method needs to use alkaline conditions to germinate the spores in vitro due to the germination method to purify the sporozoites of Bombyx mori, part of the alkali-soluble proteins located on the sporozoites will be lost under alkaline conditions, resulting in the loss of protein information

In the germination method, a 28°C water bath is required for spore germination, and the spore viability is relatively high. Only fresh spores can guarantee germination. Once diseased silkworm tissue materials or purified spores are refrigerated at 4°C for a period of time, the germination rate of spores will be very high. Low, and there is a certain ratio between spores and germination liquid. If there are too many spores, germination will be inhibited, and the germination process needs to be mixed manually at intervals, and the operation is complicated

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