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Rapid screening instrument for viable bacterium DNA, and use method and application thereof

A live-bacteria, fast technology, applied in microorganism-based methods, biochemical equipment and methods, bioreactors/fermenters for specific purposes, etc. problems, to achieve the effect of low heat output, flexible processing conditions, and improved processing efficiency

Pending Publication Date: 2021-05-07
SHANGHAI OCEAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, PMA pretreatment includes two key steps of dark treatment and photocrosslinking. The traditional PMA pretreatment method usually carries out dark treatment in a closed environment, and carries out photocrosslinking with a sodium-tungsten lamp of about 600W (such as figure 1 As shown), the process is cumbersome, and the whole manual operation is time-consuming and labor-intensive; although the photocrosslinking instrument based on blue LED tube (produced by PMA company) can greatly simplify the pretreatment process of PMA, it combines the two steps of dark treatment and photocrosslinking Separate processing still requires cumbersome manual transfer operations

Method used

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  • Rapid screening instrument for viable bacterium DNA, and use method and application thereof
  • Rapid screening instrument for viable bacterium DNA, and use method and application thereof
  • Rapid screening instrument for viable bacterium DNA, and use method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] use as figure 2 The live bacteria DNA rapid screening instrument shown quantitatively detects Vibrio in vannamei, and the steps are as follows:

[0046] (1) Construction of live bacteria DNA rapid screening instrument

[0047]LED blue lights were purchased from CREE company in the United States, a total of 24 pieces, the power is 1-3w, the voltage parameter is 3.0-3.7V, the current range is 350-1000mA, the color temperature wavelength is 410-420nm blue-medium purple / dark blue light; The explicit timer was purchased from Wenzhou Dahua Instrument Co., Ltd.; the digital display rotary regulator was purchased from China Taiwan Jingyan Electric Co., Ltd. (JSCC); the transformer was purchased from China Taiwan Mingwei Co., Ltd.; the welding equipment used argon arc The welding machine was purchased from Shenzhen Ruilong Welding Machine, product model: TIG400GT; the stainless steel plate was purchased from Xiamen Tungsten Industry, 1.5mm, food grade 304.

[0048] (2) Vibrio...

Embodiment 2

[0081] This embodiment is basically the same as Embodiment 1, except that the applied samples are Pacific oysters purchased at the aquatic product market in Pudong New Area, Shanghai. PMA double qPCR technology standard curve see Figure 4 , where the R of the standard curve of Vibrio parahaemolyticus 2 0.999, the slope of the standard curve is -3.47, the amplification efficiency is 94.17%, the R of the Vibrio cholerae standard curve 2 is 0.975, the slope of the standard curve is -3.47, and the amplification efficiency is 93.07%. The results are shown in Table 4.

[0082] Table 4: The ability of PMA duplex qPCR technology to distinguish dead and live bacteria in Pacific oysters

[0083]

[0084] Note: VP: Vibrio parahaemolyticus; VC Vibrio cholerae.

Embodiment 3

[0086] This embodiment is basically the same as Embodiment 1, except that the applied sample is the water body of the prawn breeding environment collected from Jixian Shrimp Industry, Fengxian District, Shanghai. PMA double qPCR technology standard curve see Figure 4 , where the R of the standard curve of Vibrio parahaemolyticus 2 0.988, the slope of the standard curve is -3.49, the amplification efficiency is 93.43%, the R of the Vibrio cholerae standard curve 2 is 0.994, the slope of the standard curve is -3.49, and the amplification efficiency is 90.94%. The results are shown in Table 5.

[0087] Table 5: The ability of PMA dual qPCR technology to distinguish dead and live bacteria in shrimp culture environment water

[0088]

[0089] Note: VP: Vibrio parahaemolyticus; VC Vibrio cholerae.

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Abstract

The invention discloses a rapid screening instrument for viable bacterium DNA, and a use method and application thereof. The screening instrument comprises a PMA top cover, LED blue light sources, a cuboid box type structure, a digital display type timer, a digital display type rotary regulator and a rotary mixer, wherein the cuboid box type structure is formed by fixedly connecting an upper box body with a groove structure and a lower box body with a built-in motor into a whole; the digital display type timer, the digital display type rotary regulator and the rotary mixer are arranged in the upper box body; the rotary mixer is fixed on the inner side of the groove of the upper box body in parallel and can rotate along the central axis of the groove; a plurality of sample placing holes are uniformly distributed in the rotary mixer; the plurality of LED blue light sources are uniformly distributed in the upper box body; the PMA top cover is of an L-shaped flat plate structure which is made of a non-transparent material and is subjected to edge sealing by rubber; and one side of the PMA top cover is fixed on one side of the opening of the groove of the upper box body, and the other side of the PMA top cover can be freely opened and closed along the other side of the opening of the groove of the upper box body. According to the rapid screening instrument, dark treatment and photo-crosslinking are combined into a whole, so that integration and full automation of a PMA pretreatment process are realized; the rapid screening instrument can be used for quantitative detection of viable bacteria of vibrio parahaemolyticus and vibrio cholerae in aquatic products.

Description

technical field [0001] The invention relates to the technical field of microbial analysis equipment, in particular to a rapid DNA screening instrument for live bacteria, its use method and its application in quantitative detection of Vibrio in aquatic products. Background technique [0002] Vibrio (Vibrio) is the most common pathogenic bacteria in aquatic products, and Vibrio parahaemolyticus (Vibrioparahaemolyticus) is the main pathogenic bacteria in aquatic products in my country, and the food poisoning incidents caused by this bacteria rank first in bacterial food poisoning incidents. It is also considered one of the most important causes of diarrheal diseases worldwide. Vibrio cholerae (Vibrio cholerae) is the main source of cholera disease in the human body. It is widely prevalent and is one of the severe infectious diseases. How to quickly, accurately and efficiently detect pathogenic Vibrio in aquatic products is of great significance. [0003] The traditional method...

Claims

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Application Information

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IPC IPC(8): C12M1/36C12M1/34C12M1/10C12M1/00C12Q1/6806C12Q1/689C12Q1/06C12R1/63
CPCC12Q1/6806C12Q1/689C12Q2523/101C12Q2563/173C12Q2531/113
Inventor 张昭寰黄振华赵勇童金蓉吴倩
Owner SHANGHAI OCEAN UNIV