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PCR (polymerase chain reaction) Detection method for Enterocytozoon hepatopenaei infection in penaeid shrimps

A shrimp hepatic Enterocystis and detection method technology, which is applied in the field of detection of marine biological pathogens, can solve the problem of low sensitivity and achieve the effect of simplified operation steps, simple operation, and easy operation

Inactive Publication Date: 2018-08-07
LUDONG UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to solve the problem that the sensitivity of the existing PCR detection method is not high, to provide a PCR detection kit for shrimp infection with Enterocystis hepatica, and to provide a detection method using the kit for detection, which has high sensitivity and specificity OK, simple and fast

Method used

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  • PCR (polymerase chain reaction) Detection method for Enterocytozoon hepatopenaei infection in penaeid shrimps
  • PCR (polymerase chain reaction) Detection method for Enterocytozoon hepatopenaei infection in penaeid shrimps
  • PCR (polymerase chain reaction) Detection method for Enterocytozoon hepatopenaei infection in penaeid shrimps

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specific Embodiment approach

[0025] The specific embodiment: the specific embodiment of the present invention will be further described below in conjunction with accompanying drawing:

Embodiment

[0026] Embodiment: Shrimp Infected with Shrimp Enterocystis PCR Detection Kit (see image 3 ), including a box body 1, a box cover 2 arranged at the opening of the box body 1, the rear end of the box cover 2 is rotationally connected with the opening of the box body 1, and the front end is bent downward to form a connecting portion 9, the connecting portion 9 An iron sheet 10 is provided towards the inner side of the box body 1, and a magnet corresponding to the iron sheet 10 is provided at the front end of the box body 1. A support plate frame 3 is provided inside the box body 1, and a PCR buffer is provided on the support plate frame 3. Tube hole, primer tube hole, deionized water tube hole, negative control tube hole, positive control tube hole, PCR buffer tube 4 is provided at the PCR buffer tube hole, and 2×PCR buffer is installed inside; primers are provided at the primer tube hole Tube 5, containing forward and reverse primers of Enterocystoma hepatica, forward primer s...

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Abstract

The invention discloses a PCR (polymerase chain reaction) detection method for Enterocytozoon hepatopenaei infection in penaeid shrimps and a detection kit. The method comprises: (1) designing primers; (2) providing a PCR system; (3) carrying out PCR. The detection kit includes a box body and a box cover; a PCR buffer tube filled with 2*PCR buffer is rested on a support plate; a primer tube filledwith forward and revers primers of Enterocytozoon hepatopenaei, wherein a forward primer sequence is 5'-ATT AGA CAC CGC TGT AGT TC-3', and a reverse primer sequence is 5'-GTT ATT GCC TTC TCC CTC T-3'; a deionized water tube filled with aseptic deionized water; a negative control tube filled with SPF (specified-pathogens free) nucleic acid of penaeid shrimp; a positive control tube filled with 106-copy cDNA of Enterocytozoon hepatopenaei. The PCR (polymerase chain reaction) detection method for Enterocytozoon hepatopenaei infection in penaeid shrimps and the detection kit have the advantages of high sensitivity, good specificity, good operational simplicity and high speed.

Description

Technical field: [0001] The invention relates to a PCR detection method and a detection kit for shrimp infected with Enterocystis hepatica, belonging to the technical field of detection of marine biological pathogens. Background technique: [0002] Shrimp Enterococcus hepatica ( Enterocytozoon hepatopenaei , EHP) was cultured in Thailand in 2009 in slow-growing P. monodon ( Penaeus monodon ), mainly infecting Penaeus monodon and Litopenaeus vannamei ( Litopenaeus vannam ei ), is now widely popular in Thailand, Vietnam, China, Indonesia, Malaysia and other countries. EHP is an intracellular parasite belonging to the family Microsporidiaceae and the genus Enterococcus. It mainly parasitizes in the hepatopancreas of prawns, causing growth retardation of prawns and causing huge economic losses to the prawn farming industry. [0003] Shrimp infected with EHP has no obvious appearance symptoms. Sensitive, specific and rapid detection methods are important and effective means ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12Q1/686
CPCC12Q1/686C12Q1/6888C12Q2563/107C12Q2545/114
Inventor 闫冬春庞能源粘艳莺陈博堃刘云浩王佳翠
Owner LUDONG UNIVERSITY
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