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Probe, kit and detection method for rapidly detecting nosema bombycis

A microsporidia and detection method technology, which is applied in the field of probes for rapid detection of silkworm microsporidia, can solve the problems of unfavorable and timely prevention and control, and achieve the effects of reducing reagent mixing and sample addition steps, rapid detection, and high sensitivity

Active Publication Date: 2022-04-05
广西壮族自治区蚕业技术推广站
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

When the female moth is infected with microsporidia, it can be transmitted vertically to the offspring through the embryo seed. The current general practice of quarantine of finished eggs is to judge whether the silkworm eggs are qualified or not by microscopically examining the female moth for microsporidia. The standard is set based on the average embryo transmission rate of 30% of silkworm microparticulate disease, and the actual situation of the finished egg carrying the virus needs further research
In the silkworm stage, silkworm microparticle disease can only be visually identified in the middle and late stages of infection, which is not conducive to the timely prevention and control of the disease

Method used

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  • Probe, kit and detection method for rapidly detecting nosema bombycis
  • Probe, kit and detection method for rapidly detecting nosema bombycis
  • Probe, kit and detection method for rapidly detecting nosema bombycis

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Embodiment Construction

[0041] The present invention will be further described in detail below in conjunction with the embodiments, so that those skilled in the art can implement it with reference to the description.

[0042]

[0043] 1. Synthesis of probes and primers

[0044] Since the length of the probe sequence needs to be between 40-50bp, and the quenching group and the fluorescent group are respectively connected to a pair of thymine nucleotides, the pair of oligo-Ts are separated by 1-5 bases , one of the heterozygous 1-5 bases is replaced by tetrahydrofuran, designed according to the full-length 762bp of the EB1-Probe gene published by GenBank, the probe starts at 216-256bp, a total of 41bp, quencher group and fluorescence The groups are respectively connected at positions 241 and 245, and thymine at position 242 is replaced by tetrahydrofuran to form the desired detection probe (EB1-Probe). The nucleotide sequence of EB1-Probe is: TGAATTGTATTTTCCAGTAGAGAAA / i6FAMdT / [THF]AA / Based on iBHQ1...

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Abstract

The invention discloses a probe for rapidly detecting nosema bombycis. The nosema bombycis can be rapidly detected through an RAA technology. The invention further discloses a kit for rapidly detecting the nosema bombycis, and the kit is rapid in detection and high in sensitivity. The invention further discloses a detection method of the kit for rapidly detecting the nosema bombycis, the steps of reagent mixing and sample adding are reduced, operation is convenient, and time is saved.

Description

technical field [0001] The invention relates to the field of biotechnology, more specifically, to a probe, a kit and a detection method for rapidly detecting the microsporidia of silkworm. Background technique [0002] No. silkworm, belonging to the Kingdom Fungi, is a type of intracellular obligate parasitic eukaryote. According to its ability to parasitize in silkworms, it is named "No. silkworm". When the female moth is infected with microsporidia, it can be transmitted vertically to the offspring through the embryo seed. The current general practice of quarantine of finished eggs is to judge whether the silkworm eggs are qualified or not by microscopically examining the female moth for microsporidia. The standard is set based on the average embryo transmission rate of 30% of silkworm microparticulate disease, and the actual virus-carrying condition of finished eggs needs further research. In the silkworm stage, the silkworm microparticle disease can only be visually dis...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12Q1/6844C12N15/11C12R1/645
CPCY02A50/30
Inventor 潘志新唐亮胡文娟蒋满贵唐明艳董桂清赵烨芸陈小青黄深惠
Owner 广西壮族自治区蚕业技术推广站
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