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A method for rapid detection of Listeria monocytogenes and Staphylococcus aureus

A technology for Listeria monocytogenes and Staphylococcus, which is applied in the field of rapid detection of Listeria monocytogenes and Staphylococcus aureus in food, and can solve problems such as cumbersome operations and steps, endangering consumer safety, and low detection rate , to achieve the effect of reducing detection time, avoiding false negative phenomenon, and simple operation

Active Publication Date: 2022-04-12
HENAN BUSINESS SCI RES INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The traditional national standard detection method (GB 4789.10 and GB 4789.30) takes at least 2-5 days to produce results, and requires bacteria enrichment, culture medium cultivation, separation, detection, etc. The operation and steps are cumbersome, time-consuming, and the separation rate is low, and it can only be detected The detection rate of the surviving bacteria in the sample is lower than that in the actual sample, and false negative results are prone to occur, endangering the safety of consumers

Method used

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  • A method for rapid detection of Listeria monocytogenes and Staphylococcus aureus
  • A method for rapid detection of Listeria monocytogenes and Staphylococcus aureus
  • A method for rapid detection of Listeria monocytogenes and Staphylococcus aureus

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] The screening of embodiment 1 BASV concentration

[0032] 1.1 Listeria monocytogenes

[0033] Take 225mL of PBS buffer solution containing different concentrations of BASV (0.02mol / L in PBS) in 6 parts respectively in homogenization bags, add 25g samples containing Listeria monocytogenes to each homogenization bag and homogenize for 30s, add The concentrations of BASV were 0% (no BASV added), 0.5%, 1%, 5%, 8% and 10%.

[0034] In addition, 225 mL of Listeria monocytogenes broth LB1 was taken as a comparison solution, and 25 g of a sample containing Listeria monocytogenes was added and homogenized for 30 seconds to prepare a sample solution as a comparison example.

[0035] According to the traditional detection method, operations such as enrichment, streak separation, and identification were performed on the above sample liquid, and the test results were obtained. The results are shown in Table 1.

[0036] Staphylococcus aureus

[0037] With reference to the above-me...

Embodiment 2

[0041] Example 2 Optimum Addition of Growth Solution

[0042]Take 25g of positive samples containing Listeria monocytogenes and Staphylococcus aureus respectively, add them into a homogenization bag containing 1% BASV in 225mL PBS buffer, and homogenize for 30s. Take 6 parts of 5mL sample bacterial liquid, collect the bacterial cells after centrifugation at 4°C and 8000rpm, add 0.1mL, 0.5mL, 1mL, 2mL, 4mL, 5mL of growth solution, and mix well. Then smear it on the plates corresponding to the two kinds of bacteria respectively, carry out operations such as streak separation and biochemical test according to GB4789, and obtain the test results, as shown in Table 2. The whole process detection method of GB4789 is also used as a comparison in the table.

[0043] Table 2 The results of colony detection with different amounts of growth solution added

[0044]

[0045] It can be seen from the table that with the increase of the growth liquid, the number of bacteria also increases...

Embodiment 3

[0046] Example 3 Optimum time for ultrasonic oscillation mixing

[0047] Take 25 g of positive samples containing Listeria monocytogenes and Staphylococcus aureus, add them to homogenizing bags containing 225 mL of PBS buffer containing 1% BASV, and homogenize for 30 seconds. Take 5 parts of 5mL sample bacterial liquid, collect the bacterial cells after centrifugation at 4°C and 8000rpm, add 1mL growth liquid, mix the growth liquid with the bacterial liquid, and ultrasonically oscillate respectively. The oscillation time of the five samples is 1min, 3min, 5min, and 8min in turn. and 10min, after shaking, spread on the plates corresponding to the two kinds of bacteria to carry out operations such as streak separation and culture, and the detection results are shown in Table 3.

[0048] Table 3 The number of colonies detected by ultrasonic oscillation at different times

[0049]

[0050] The results showed that when the shaking mixing time was 5 minutes, the number of coloni...

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Abstract

The invention discloses a method for rapidly detecting Listeria monocytogenes and Staphylococcus aureus, belonging to the field of biological detection. The steps of the method include: (1) taking a sample under sterile conditions, adding it to a phosphate buffer solution containing the fifth component of bovine serum albumin and homogenizing the sample to obtain a homogeneous solution; (2) centrifuging the sample homogenized solution at low temperature to collect bacteria Add growth medium to the collected bacteria and shake to form a bacterial suspension. Take immunomagnetic beads and mix evenly with the bacterial suspension. After heat treatment in a water bath, put them on a magnetic stand for stratification. Discard the supernatant and add the eluent. Wash the lower layer of the sample to obtain the sample solution to be tested, apply it on a plate for cultivation, observe and detect. The detection method provided by the invention can effectively shorten the detection time, reduce the time cost, and find and cut off the source of infection in time.

Description

technical field [0001] The invention belongs to the technical field of microorganism detection, and in particular relates to a method for rapidly detecting Listeria monocytogenes and Staphylococcus aureus in food. Background technique [0002] Listeria monocytogenes is a zoonotic food-borne pathogen widely distributed in nature. It can cause listeriosis in humans and animals. After infection, the main manifestations are sepsis, meningitis, and mononucleosis. Serious harm to pregnant women, newborns, the elderly and immunocompromised patients. This bacterium can widely exist in a variety of foods, such as meat products, dairy products, aquatic products, vegetables, etc. This bacterium has caused many large-scale food poisonings in European and American countries, so the rapid growth of Listeria monocytogenes in food Detection is very necessary, and effective control of Listeria monocytogenes in food is one of the important issues of food safety. [0003] Staphylococcus aure...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/14C12Q1/04
CPCC12Q1/04C12Q1/14G01N2333/195G01N2333/31
Inventor 周莉张立攀章建军关炳峰朱海华李向力平洋张亚勋谭静高火亮王慧张文杰陈红赵梦瑶王晓瑞
Owner HENAN BUSINESS SCI RES INST
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