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Test chip for fast detecting microorganism, preparation and use thereof

A technology of test strips and microorganisms, applied in the direction of microorganism-based methods, microorganism measurement/inspection, biochemical equipment and methods, etc., can solve the problems of slow water absorption, low recovery rate, fusion of gels, etc., to avoid colony omission , Simple operation, easy to carry

Inactive Publication Date: 2009-08-05
GUANGDONG INST OF MICROORGANISM
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The test piece with filter paper as the carrier is simple to manufacture and the material is cheap, but the filter hole is too large, and it is easy to have colonies growing on both sides and cannot be counted accurately; for the detection of oily food, the filter paper absorbs slowly, and it is difficult to diffuse to make the sample liquid evenly distributed; The test piece with non-woven fabric as the carrier has the advantage of quickly and evenly spreading the sample liquid, but because of its opaqueness and thick non-woven fabric layer, the colonies in the lower layer of the carrier cannot be counted, resulting in a slightly lower recovery rate
The petrifilm gel test sheet of 3M Company in the United States, because the top layer of the sheet is covered with a thin film, when the bacteria produce gas and mucus are too much, the phenomenon of colony diffusion and fusion will affect the count; in addition, because the gel absorbs water slowly, the sample The liquid is easy to spill out of the range of the medium, affecting the recovery rate
Currently, the test strips on the market are opaque, cannot count bacteria on both sides to avoid omissions, and are difficult to count through automatic bacteria counters; in addition, except for 3M’s petrifilm test strips, all kinds of products cannot be used for next-step identification of bacteria

Method used

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  • Test chip for fast detecting microorganism, preparation and use thereof
  • Test chip for fast detecting microorganism, preparation and use thereof
  • Test chip for fast detecting microorganism, preparation and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Embodiment 1: the preparation of the present invention's rapid determination of the total number of microbial colonies test piece

[0040] Include the following steps:

[0041] 1) Cut each part, the length and width of the plastic film are 10.5×7.5cm; the length and width of the bottom plate and the film are both 9.5×7.5cm; the bottom surface of the hollow film in the middle is glued to the bottom plate to form a circular groove cultivation area with a diameter of 6cm;

[0042] 2) Mix 6 g of nutrient agar powder required for microbial growth and 0.2 g of TTC, and bond 0.06 g of the mixed powder in the groove culture area through a pressure-sensitive adhesive;

[0043] 3) heat the polyacrylic resin pressure-sensitive adhesive into a liquid, evenly spray it on the bottom surface of the plastic film, and bond 0.3g of cold water condensable gel powder to the bottom surface of the plastic film;

[0044] 4) The bottom surface of the plastic film and the upper surface of the ...

Embodiment 2

[0047] Embodiment 2: Utilize embodiment 1 test sheet to measure the total number of colonies, and compare with the international plate pouring method Coccus (Staphylococcus aureus ATCC 6538), Staphylococcus aureus (Staphylococcus aureus CMCC 26003), and Pseudomonas aeruginosa (Pseudomonas aeruginosa ATCC 9027) were inoculated into 5ml of nutrient broth, cultured at 37°C for 18-24h, each Take 1ml in a sterile test tube to obtain a mixed bacterial solution, and adjust the concentration to about 10 5 cell / ml and dilute to 10 times with normal saline 4 、10 3 、10 2 、10 1 、10 0 and 10 -1 cell / ml, three replicates were set for each concentration gradient, and they were set aside.

[0048] 2) Turn over the plastic film on the upper layer of the test piece, drop 1ml of the mixed bacterial solution of each concentration gradient into the groove culture area, cover the plastic film, observe whether the sample liquid has been evenly dispersed in the center of the test piece, let it ...

Embodiment 3

[0052] Embodiment 3: Utilize the test piece of embodiment 1 to visually observe the number of different bacterial species colonies, and compare with the automatic colony counter measurement result

[0053] 1) Preparation of bacterial liquid: Escherichia coli (Escherichia coli 8099), Salmonella typhimurium (Salmonella typhiCMCC 50115), Staphylococcus aureus (Staphylococcus aureus ATCC 6538), Staphylococcus aureus (Staphylococcus aureus CMCC 26003), aeruginosa Pseudomonas aeruginosa ATCC 9027 was inoculated into 5ml of nutrient broth, cultured at 37°C for 18-24h, each 1ml was taken in a sterile test tube, and diluted 10 times with normal saline until the concentration of each bacterial solution was 10 1 Cell / ml, set up two replicates for each bacterial strain, set aside.

[0054] 2) Take five test pieces and measure the five bacterial solutions in step 1) respectively. Open the plastic film on the upper layer of the test piece, and add a concentration of 10 1 1ml of cell / ml ba...

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Abstract

The invention discloses a testing piece for quickly testing microorganisms, as well as a preparation method and application thereof. The testing piece comprises a soleplate, a film with a hollow part in the middle and a plastic film. The bottom surface of the film with the hollow part in the middle is adhered to the soleplate; a groove culture area is formed at the hollow part in the middle of the film; a color development culture medium is arranged in the groove culture area; a cold water condensable gellant is adhered to the bottom surface of the plastic film; and the bottom surface of the plastic film and the upper surface of the film are adhered to the edge at one side. After the upper and the lower layers of the testing piece are adhered, irradiation and sterilization packages are used for standby. The testing piece can be applied to the detection of total colony count, coliform bacteria, coliform group, salmonella, staphylococcus aureus, subsidiary haemolytic vibrio, pseudomonas aeruginosa and other food-borne causative organisms and has the advantages of simple operation, convenient carrying, short testing time, accurate result and double-sided bacteria count.

Description

technical field [0001] The invention relates to the field of food microbiology safety inspection and detection, in particular to a test piece for rapid detection of microorganisms and a preparation method and application thereof. Background technique [0002] The current national food hygiene microbiological testing standard method has cumbersome operating steps. Before and after testing microbes, it is necessary to prepare culture medium, clean culture vessels, sterilize, enrich bacteria, and take a long time to culture. At the same time, it has high requirements for experimenters and laboratories. Under today's ever-accelerating pace of commercial trade, traditional detection methods can no longer meet the detection needs of food production and operation enterprises, import and export commodity inspection, and government management departments. [0003] The test piece uses paper, cold water coagulable gel and non-woven fabric as the medium carrier to measure the content of...

Claims

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Application Information

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IPC IPC(8): C12Q1/04G01N21/78C12R1/19C12R1/42C12R1/445C12R1/63C12R1/385
Inventor 吴清平吴许文黄汝添张菊梅蔡芷荷郭伟鹏
Owner GUANGDONG INST OF MICROORGANISM
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