Method for quickly quantifying concentration of trehalose in single microsporidia spore

Abstract

The invention discloses a method for quickly quantifying the concentration of trehalose in a single microsporidia spore based on a single cell Raman spectrum, and belongs to the technical field of quantitative analysis. The method mainly adopts a single cell Raman spectrum technology and a pair of laser tweezers to quantitatively analyze the concentration of the trehalose in the single microsporidia spore. The method is easy and convenient to operate and pollution-free, and no chemical agents are needed; a spore sample is directly detected in germfree water, and the concentrations of the trehalose in the individual microsporidia spores from different sources can be quickly and intuitively quantified, so that heterogeneity of the concentration of the trehalose in the single spore in the same colony is obtained; the method has the advantages of simplicity, efficiency and reliability.

Description

technical field [0001] The technical field of quantitative detection of the present invention specifically relates to a method for directly quantitatively analyzing the trehalose content of Microsporidia spores based on single-cell Raman spectroscopy. [0002] technical background [0003] Microsporidia (Microsporidia) is a kind of intracellular obligate parasitic unicellular eukaryotes, which are currently classified as fungi, with a wide range of hosts, and can infect invertebrates and vertebrates including humans. There are more than 150 genera and more than 1400 species. Microsporidia are pathogenic to many economically valuable insects, fish, rabbits, etc. Among them, Nosema bombycis (N.b) is the earliest microsporidia discovered by humans, and it mainly parasitizes silkworms. Bombyx mori microsporidia is a kind of contagious silkworm disease caused by N.b infection and parasitism. It can be transmitted through ingestion or embryo seed infection. It is a devastating di...

AI Technical Summary

Problems solved by technology

Trehalose enzymatic analysis is a recently developed assay method. Its principle is to specifically hydrolyze trehalose by trehalase, and then detect the glucose produced by the glucose oxidase-peroxidase system. This method has high specificity. , the analysis cost is also high, and it is affected by enzyme activity

[0006] The above methods are all time-consuming, reagent-consuming and sample-consuming detection methods. These methods need to break the spore wall first, release trehalose from the spores, and require a large number of spore samples to meet the minimum sample requirements for detection, so they cannot Quantitative trehalose concentration of microspores, let alone individual spores

Moreover, the spore samples after detection are destroyed and have no cell activity, so it is impossible to directly analyze the correlation between the concentration of trehalose and the germination and infectivity of spores.

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