Method for detecting Nosema bombycis spores in lepidopteran insects, staining solution and preparation method thereof

A lepidopteran insect and detection method technology, applied in the detection method of lepidopteran insect microparticle protozoan spores and its staining solution and preparation field, can solve the problems of complicated operation, high cost, and difficulty in wide application, and achieve easy operation , easy to identify, high practical value effect

Inactive Publication Date: 2019-05-07
广西壮族自治区蚕业技术推广总站
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Although the method has high accuracy, the operation is complicated...

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  • Method for detecting Nosema bombycis spores in lepidopteran insects, staining solution and preparation method thereof
  • Method for detecting Nosema bombycis spores in lepidopteran insects, staining solution and preparation method thereof

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Embodiment Construction

[0022] 1. Inspection of Bombyx moth microparticles and protozoan spores

[0023] The silkworm is an insect that feeds on mulberry leaves, and belongs to the order Insecta, Lepidoptera, Silkworm family, and the genus Bombyx mori. The scientific name is Bombyxmori Linnaeu. Its first generation has four stages: egg, larva, pupa, and moth.

[0024] Use isothiocyanate powder as fluorescent staining agent, dissolve isothiocyanate powder in phosphate buffer solution with a pH value of 7.8 to a concentration of 0.1mg / mL, then filter through a filter (mesh 35-45μm), and store in the refrigerator Store in liquid state at -4 to 5°C in the dark.

[0025] The sample is Bombyx mori after laying eggs. Take the sample moth and add 0.5% NaOH solution to grind, and get the purified standard product Bombyx mori microparticle protozoan spore (scientific name: Nosema bombycis Linnaeu) belongs to Fungi (Fungi), Microspora (Microspora), Microsporea (Microsporea) ), about 0.005g of Microsporida (...

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Abstract

The invention discloses a staining solution for Nosema bombycis spores in lepidopteran insects, which is a phosphate buffer containing isothiocyanate. Accordingly, the inventor also establishes a method for preparing the staining solution, and a detection method using the staining solution. In the detection method, a sample is fluorescently stained by the above staining solution, and then the spores are observed under a fluorescence microscope. The staining solution is used to fluorescently stain shells of the Nosema bombycis spores in the lepidopteran insects, and slides are easy to manufacture and easy to operate. The fluorescence microscope is used, and the staining solution used for staining the Nosema bombycis spores in the lepidopteran insects makes the Nosema bombycis spores in thelepidopteran insects have fluorescence specificity. The repeatability and accuracy of the detection are ensured by comparing with shape characteristics of a standard. In summary, the method for detecting the Nosema bombycis spores in the lepidopteran insects, the staining solution and the preparation method thereof have the characteristics of simple operation, strong specificity and easy identification, and lay a foundation for mechanical detection and fair detection, and have high practical value and broad market prospect.

Description

technical field [0001] The invention belongs to the field of lepidopteran insect detection and quarantine, and in particular relates to a detection method of lepidopteran insect microparticle protozoa spores, a staining solution and a preparation method thereof. Background technique [0002] Among the Lepidoptera insects, some are economic insects, such as silkworms and tussah silkworms. However, Lepidoptera insects can be infected with microparticle protozoon diseases, such as silkworm microparticle protozoon disease, tussah silkworm microparticle protozoon disease, castor silkworm microparticle protozoon disease, etc. Particulate protozoa are single-celled organisms that parasitize within cells. There are two main routes of infection: ingestion and germ infection. Particulate protozoa cause ingestion through host excrement and corpses; infection occurs through host germ embryos. The next generation is called germ-to-seed infection. [0003] Since the microparticle protoz...

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Application Information

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IPC IPC(8): G01N21/64
Inventor 朱方容黄深惠黄旭华李韬王霞蒋满贵李标
Owner 广西壮族自治区蚕业技术推广总站
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