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LAMP detection kit for Enterocytozoon hepatopenaei

A detection kit and technology for microsporidia, which are used in the determination/inspection of microorganisms, resistance to vector-borne diseases, biochemical equipment and methods, etc., can solve the problem that the detection method cannot be directly applied, and achieve high sensitivity and good effect. , the effect of high detection sensitivity

Inactive Publication Date: 2017-04-26
ZHEJIANG INST OF FRESH WATER FISHERIES
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Problems solved by technology

[0005] The purpose of the present invention is to provide a detection kit for Microsporidia hepatica LAMP, which has strong specificity, high sensitivity, simple and quick operation, overcomes the problem that existing detection methods cannot be directly applied in the monitoring of breeding diseases, and is suitable for shrimp Screening and prevention of hepatic enteric microsporidiosis

Method used

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  • LAMP detection kit for Enterocytozoon hepatopenaei
  • LAMP detection kit for Enterocytozoon hepatopenaei
  • LAMP detection kit for Enterocytozoon hepatopenaei

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Embodiment

[0038] A detection kit for Microsporidia hepatica LAMP, comprising tissue lysate, LAMP amplification reaction solution, a positive control sample and a negative control sample, wherein the positive control sample is Microsporidia hepatica genomic DNA, and the negative control sample is nucleic acid-free RNase free water.

[0039] The components of the LAMP amplification reaction solution are as follows:

[0040] Primer EHP-FIP 1.4mM, Primer EHP-BIP 1.4mM, Primer EHP-F3 0.2mM, Primer EHP-B30.2mM, 20mM Tris-HC (pH 8.0~9.0), 10mM Potassium Chloride, 15mM Ammonium Sulfate, 8mM Sulfate Magnesium, 0.1% TritonX-100, 0.6M betaine, 1.4mM dNTP, 2.5U / μL Bst DNA polymerase large fragment, 1.25mM calcein, 25mMMnCl2, the balance of water.

[0041] Primer EHP-F3: 5'-GGGATCAAGGACGAAGGCT-3' (SEQ ID No.1);

[0042] Primer EHP-B3: 5'-CAGCACAATCCACTCCTGG-3' (SEQ ID No. 2).

[0043] Primer EHP-FIP:

[0044] 5'-AAGCATCGCTCTCGCAACACCACACCGCTGTAGTTCTAGCA-3' (SEQ ID No. 3),

[0045] Primer EHP-BI...

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Abstract

The invention discloses an LAMP detection kit for Enterocytozoon hepatopenaei. The LAMP detection kit comprises a tissue lysate and an LAMP amplification reaction solution, wherein the LAMP amplification reaction solution comprises 1.4 mM LAMP inner primer, 0.2 mM LAMP outer primer, 20 mM Tris-HC, 10 mM potassium chloride, 15 mM ammonium sulfate, 8 mM magnesium sulfate, 0.1% Triton X-100, 0.6 M betaine, 1.4 mM dNTP, 2.5 U / [mu]L Bst DNA polymerase large fragment, 1.25 mM calcein, 25 mM MnCl2, and the balance of water. The detection kit of the present invention has advantages of strong specificity, high sensitivity and quick and easy operation, can overcome the incapable application problem of the existing detection method in the breeding disease monitoring, and is suitable for the screening and the prevention of Enterocytozoon hepatopenaei.

Description

technical field [0001] The invention relates to the technical field of aquatic animal disease detection, in particular to a LAMP detection kit for Shrimp Hepatenterentericus LAMP. Background technique [0002] Shrimp hepatopenaei (Enterocytozoon hepatopenaei, EHP) disease is caused by a microsporidia parasitism, named according to the symptoms manifested in Thailand monodon prawn (Penaeus monodon) in 2009, mainly present in shrimp In the hepatopancreas (HP), EHP outbreaks have been widely occurring in China, Indonesia, India, Malaysia, and Thailand since 2014. Therefore, EHP is an emerging problem among farmed shrimp diseases and has caused great harm to the global shrimp industry. , urgently need control. Studies have found that EHP can infect Penaeus vannamei, Macrobrachium rosenbergii, Macrobrachium japonica, Penaeus monodon, etc., and can be transmitted both vertically and horizontally. The direct transmission mode between shrimp and shrimp may be It is transmitted ora...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/04
CPCC12Q1/6844C12Q1/6893C12Q2531/119C12Q2563/107Y02A50/30
Inventor 林锋刘莉曹铮卢淑娟
Owner ZHEJIANG INST OF FRESH WATER FISHERIES
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