Method for detecting Y. enterocolitica

A technology for Yersinia and enterocolitis, applied in the field of microbial detection, can solve the problems of complicated operation steps, inability to achieve rapid detection, time-consuming, etc., to reduce interference, improve detection rate and accuracy of detection results, reduce The effect of false positives

Active Publication Date: 2019-04-30
HENAN BUSINESS SCI RES INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The conventional detection method of Yersinia enterocolitica is complex and time-consuming, generally takes 3 to 7 days, which cannot meet the requirements of rapid detection of Yersinia enterocolitica in food

Method used

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  • Method for detecting Y. enterocolitica
  • Method for detecting Y. enterocolitica
  • Method for detecting Y. enterocolitica

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1: Al(OH) 3 concentration screening

[0037] Take 5 parts of 225ml containing different concentrations of Al(OH) 3 PBS buffer solution (concentration of PBS buffer solution is 0.02mol / L) in five sterile homogeneous bags (Al(OH) in five homogeneous bags 3 The mass concentration is respectively 0.5%, 1%, 3%, 5%, 10%), and 25g Yersinia enterocolitica positive sample is added in each homogeneous bag (specifically adopting Yersinia enterocolitica containing Yersinia enterocolitica Bacteria cold chain meat), homogenized for 30s to prepare the sample bacterial liquid.

[0038] In addition, use 225ml of modified phosphate buffer without Al(OH) 3 PBS buffer solution (0.02mol / L PBS) was used as a contrast solution, and 25 g of Yersinia enterocolitica positive samples were added to each contrast solution, and homogenized for 30 seconds to prepare a sample bacterial solution.

[0039] According to the detection method stipulated in GB 4789.8, operations such as bacteria...

Embodiment 2

[0043] Embodiment 2: the screening of water bath heat treatment temperature

[0044] Get 25g of Yersinia enterocolitica positive sample (specifically cold chain meat containing Yersinia enterocolitica), add 225mL containing 3% Al(OH) 3 In a sterile homogenization bag of PBS buffer solution, homogenize for 30 seconds to obtain the sample bacterial solution. Take 5mL of sample bacterial liquid, and heat-treat them in water baths at 30°C, 40°C, 50°C, 60°C, and 80°C for 15 minutes to kill non-target bacteria. Then, according to the detection method stipulated in GB 4789.8, operations such as bacterial enrichment, streak separation, and biochemical tests were carried out in sequence, and the test results were obtained, as shown in Table 2 for details.

[0045] Table 2 The number of detected colonies of different water bath heat treatment temperatures

[0046]

[0047] It can be seen from Table 2 that when the water bath heat treatment temperature is not higher than 60°C, the n...

Embodiment 3

[0048] Embodiment 3: the screening of water bath heat treatment time

[0049] Get 25g of Yersinia enterocolitica positive sample (specifically cold chain meat containing Yersinia enterocolitica), add 225mL containing 3% Al(OH) 3 In a sterile homogenization bag of PBS buffer solution, homogenize for 30 seconds to obtain the sample bacterial solution. Take 5mL of the sample bacterial solution and heat-treat in a 60°C water bath for 5min, 10min, 15min, 20min, and 30min, respectively, to kill non-target bacteria. According to the detection method stipulated in GB 4789.8, operations such as bacterial enrichment, streak separation, and biochemical tests were carried out in sequence, and the test results were obtained. See Table 3 for details.

[0050] Table 3 The number of detected colonies of different water bath heat treatment temperatures

[0051]

[0052] It can be seen from Table 3 that when the water bath time does not exceed 15 min, the number of detected colonies increa...

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Abstract

The invention discloses a method for detecting Y. enterocolitica, which comprises the steps of (1) taking a sample to be detected under an aseptic condition, adding the sample into a PBS buffer solution containing Al(OH)3, and homogenizing to obtain a sample bacteria solution; (2) carrying out water-bath heat treatment on the sample bacteria solution, performing centrifugation, discarding the supernatant, and collecting thalluses; (3) adding Y. enterocolitica immune-magnetic beads into the thalluses, standing on a magnetic shelf for layering after oscillation, discarding the upper layer, obtaining Y. enterocolitica immune-magnetic beads attached with the thalluses, eluting the Y. enterocolitica immune-magnetic beads attached with the thalluses by an eluant, then adding the PBS buffer solution into the eluted Y. enterocolitica immune-magnetic beads, and uniformly mixing to obtain a sample solution to be detected; and (4) culturing, observing and detecting the sample solution to be detected. The detection method provided by the invention is high in specificity and sensitivity, and can effectively shorten the detection time and reduce the detection cost.

Description

technical field [0001] The invention belongs to the technical field of microorganism detection, and in particular relates to a method for detecting Yersinia enterocolitica. Background technique [0002] Yersinia enterocolitica (Y.enterocolitica) is a zoonotic pathogenic microorganism widely distributed in nature, present in raw vegetables, milk and dairy products, meat, soybean products, salads, oysters, clams and shrimp, and also in the environment, such as lakes, rivers, soil and vegetation. The bacterium has been isolated from the feces of domestic animals, dogs, cats, goats, chinchillas, mink and primates. Yersinia enterocolitica can be transmitted through food, and can cause Yersinia enterocolitica in various animals and humans. The main symptoms after infection are fever, abdominal pain, diarrhea, vomiting, arthritis, sepsis, etc. , seriously endangering human health. Because of this, many countries stipulate that this bacteria is a routine item that must be inspect...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543G01N33/569
CPCG01N33/54326G01N33/56916
Inventor 周莉李永利毛晓娜张立攀关炳峰平洋张亚勋谭静任钊王潇李栋蒋碧伟高瑞吴晶晶王春杰
Owner HENAN BUSINESS SCI RES INST
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