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Kit for detecting enterocytozoonhepatopenaei

A shrimp Enterocytoplasma hepatobilis, kit technology, applied in microorganism-based methods, microbial determination/inspection, microorganisms, etc., can solve the problems of aggravated horizontal transmission, food safety objection, and the incidence and loss of EHP, and achieves a reduction in the requirements, ensure safety, and shorten the effect of testing time

Inactive Publication Date: 2016-10-26
徐锦余 +1
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Problems solved by technology

Although it was later proved that the microsporidia are not the pathogenic microorganisms that cause the white stool of prawns, it will aggravate the possibility of its horizontal transmission
Since 2013, EHP has been detected in samples of domestically cultured Litopenaeus vannamei. Since 2014, the detection rate of EHP in Litopenaeus vannamei aquaculture in Zhejiang Province has increased rapidly, but there is no statistical report on the incidence and loss of EHP.
And there are also important objections to food safety for the rapid detection method of whether EHP is contained in aquatic food

Method used

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  • Kit for detecting enterocytozoonhepatopenaei
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  • Kit for detecting enterocytozoonhepatopenaei

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Embodiment Construction

[0029] 1Materials and methods

[0030] 1.1 Experimental materials

[0031] The positive samples of Enterosporium were collected from Xiangshan and Linhai Litopenaeus vannamei with intestinal worm samples. Sample numbers: NB2015-07, TZ2015-02. After PCR detection and sequencing, it was confirmed that they were infected by Enterosporium hepatica. ; Intestinal cytoplasmic nucleic acid extraction uses SH09-D nucleic acid extraction box or the nucleic acid extraction solution of the present invention for extraction; intestinal cyst gene cloning pMD19-T vector, Escherichia coli competent cells, plasmid recovery kit purchased from Beijing Quanshijin Bio Technology Co., Ltd., high-fidelity pfu premix and fluorescence quantitative PCR Taq enzyme premix are products of Nanjing Kangwei Century Biotechnology Co., Ltd.; PCR tubes and suction nozzles for experiments are all products of Axygen.

[0032] The fluorescence quantitative PCR instrument is Roche LC480, the PCR instrument is Hangzhou Lan...

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Abstract

The invention provides a kit for detecting enterocytozoonhepatopenaei and a method for detecting whether aquatic foods contain enterocytozoonhepatopenaei. Besides a primer pair, the kit further comprises nucleic acid extracting liquid. The nucleic acid extracting liquid is formed by 4% of sodium dodecyl sulfate (SDS) and 1.5M Tris / HCl (at the pH of 8.8). An established TaqMan real-time real-time fluorescence quantification polymerase chain reaction (PCR) method has a detection reaction sensitivity as high as 5.20*102 copies per microliter. Compared with ordinary PCR, the sensitivity of the method is 10 times higher than that of the conventional PCR.

Description

Technical field [0001] The present invention relates to a kit for shrimp enterocystis and a method for detecting whether aquatic food contains shrimp enteroplasma. Background technique [0002] Enterocytozoonhepatopenaei (EHP), also known as Enterocytozoon, is a microsporidian found in shrimps in recent years. It mainly infects important cultured shrimps such as Litopenaeus vannamei and Penaeusmonodon. In 2009, it was found in the slow growth of Penaeus monodon in Thai culture ponds. It has been reported that EHP has a higher detection rate in Penaeus monodon and Penaeus vannamei with white faeces syndrome (WFS) in Thailand and Vietnam, and is severely infected with Intestinal worm liver. Although it was later proved that the microsporidian is not a pathogenic microorganism that causes white prawns, it will increase the possibility of its horizontal transmission. Since 2013, EHP has been detected in the samples of Litopenaeus vannamei cultured in China. Since 2014, the detectio...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/04C12R1/90
CPCC12Q1/6893C12Q1/6851C12Q2531/113C12Q2563/107
Inventor 石坚骆云慧钱冬徐锦余
Owner 徐锦余
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