CN102453745A - Method for quickly detecting total bacterial count in solid food - Google Patents
Method for quickly detecting total bacterial count in solid food Download PDFInfo
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- CN102453745A CN102453745A CN2010105275604A CN201010527560A CN102453745A CN 102453745 A CN102453745 A CN 102453745A CN 2010105275604 A CN2010105275604 A CN 2010105275604A CN 201010527560 A CN201010527560 A CN 201010527560A CN 102453745 A CN102453745 A CN 102453745A
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Abstract
The invention relates to a method for quickly detecting the total bacterial count in a solid food. The invention solves the problem that the existing quick detection method for food microbes can not truly achieve quick detecting speed, low detecting cost, high detecting accuracy rate and low detection limit. The detection method comprises the following steps of: 1, weighing a solid sample and a peptone water solution and mixing uniformly by shaking; 2, sucking 1ml of sample for centrifuging; 3, taking 0.8ml of supernatant for re-centrifuging; 4, discarding 0.3ml of supernatant and mixing uniformly; 5, taking 5ul of bacterial liquid and dripping onto a glass slide for drying and dyeing; 6, washing with distilled water and wiping dry; and 7, observing by a microscope, and then calculating by a quick detection system for total bacterial count to obtain the total bacterial count. The invention has the advantages of high detecting speed, low detecting cost and high detecting accuracy.
Description
Technical field
The present invention relates to a kind of method for quick, the method for total plate count in definite a kind of rapid detection solid food of saying so.
Background technology
Food is the basic substance that the human life activity depends on for existence, is human life's the energy.Owing to contain abundant nutrition in the food, bacterium is easy to growth in food product environment, make food that change take place.Though normal plants and animal interior tissue are aseptic and since obtain with process such as manipulation in pollution, it is contaminated and grow various bacteria that food surfaces is attend regular meeting.Food microorganisms pollute the pollution that comprises Bacterial Contamination, virus and fungi and toxin thereof.Bacterium existence in the food is an important indicator estimating food quality and security.The bacterial contamination degree direct influence food safety, and the harm of HUMAN HEALTH and personnel safety (the danger side of body) is mainly shown two aspects.The one, food contamination-bacterial food poisoning of pathogenic bacterium, the 2nd, the food contamination-food of non-pathogenic bacteria putrid and deteriorated.According to estimates, in the annual hundreds of millions of food origin disease patient in the whole world, the 70%th, because the food that has eaten various microbial contaminations and drinking-water cause.
Total plate count is meant at test sample unit weight (g), volume (mL), or surface-area (cm
2) in, contained can be on certain solid medium, cultivate the sum of the bacterial colony that the back generated under certain condition.Total plate count is mainly as judging that food by the sign of bacterial contamination degree, has important health significance.
The aerobic plate count method is generally adopted in the detection of total plate count both at home and abroad, and (Aerobicplatecount APC) is applicable to mesophilic aerobic and amphimicrobian viable bacteria total number of bacterial colony.The method of total plate count is to adopt the national standard of " food sanitation Micro biological Tests ", both the method for plate culture count in the traditional detection food.This is with after the suitable dilution of testing sample warp; Mikrobe wherein fully is dispersed into individual cells; Getting a certain amount of diluted liquid is inoculated on the flat board; Through cultivating, form macroscopic bacterium colony by each unicellular growth and breeding, promptly single bacterium colony should represent one in the raw sample unicellular.The statistics colony count can converse according to its extension rate and sampling inoculum size and to contain the bacterium number in the sample.
The method of plate culture count operation is more numerous; The result need cultivate for some time and just can obtain; And the data that obtain can not be represented the bacterium whole alive that exists in the food; And only represent the bacterium that energy for growth is arranged when supplying with various nutrition at the specific physical environment, be linked to be blocky simultaneously or a plurality of cells of catenate or a unicellular bacterium colony, accuracy that influence is counted of all will producing only.
The objective of the invention is all to have such or such problem in order to solve existing food microorganisms method for quick, do not have a kind of method can really reach detection speed fast, detect that cost is low, the accuracy rate height that detects and the low problem of detectability and a kind of detection speed of proposing are fast, the detection method that detects total plate count in the solid food that cost is low, accuracy in detection is high.
Technical scheme of the present invention is: one, take by weighing the 20-30g solid sample under the aseptic condition and be dissolved in concussion mixing in back in the peptone water solution of 50-60ml; Two, absorption 1ml sample is centrifugal; Three, get 0.8ml supernatant recentrifuge; Four, discard mixing behind the 0.3ml supernatant; Five, get 5ul bacterium drop to slide glass, dry back is with the staining fluid 1-3min that dyes; Six, dry with lens wiping paper behind the distilled water flushing; Seven, with calculating through the total plate count rapid detection system after the microscopic examination, can obtain total plate count.
The present invention have detection speed fast, detect the low advantage of cost; The present invention adopts methylene blue as dye liquor, can accurately differentiate bacterium and non-bacterium composition, makes the present invention also have the high advantage of accuracy in detection.
Embodiment
Embodiment one: the method for total plate count realizes according to the following steps in this embodiment rapid detection solid food: one, take by weighing the 20-30g solid sample under the aseptic condition and be dissolved in concussion mixing in back in the peptone water solution of 50-60ml; Two, absorption 1ml sample is centrifugal; Three, get 0.8ml supernatant recentrifuge; Four, discard mixing behind the 0.3ml supernatant; Five, get 5ul bacterium drop to slide glass, dry back is with the staining fluid 1-3min that dyes; Six, dry with lens wiping paper behind the distilled water flushing; Seven, with calculating through the total plate count rapid detection system after the microscopic examination, can obtain total plate count.
Peptone, staining fluid are commercially available conventional reagent in this embodiment.
Embodiment two: what this embodiment and embodiment one were different is the peptone water solution that adopts 0.5% concentration in the step 1.Other step and parameter are identical with embodiment one.
Embodiment three: what this embodiment and embodiment one were different is under aseptic condition, accurately to take by weighing the 25g solid sample in rapid one, puts into the peptone water solution of 55ml, and on shaking table with 160r/min concussion 2min mixing.Other step and parameter are identical with embodiment one.
Embodiment four: what this embodiment and embodiment one were different is that centrifugal condition is 1000r/min in the step 2,8min.Other step and parameter are identical with embodiment one.
Embodiment five: what this embodiment and embodiment one were different is that centrifugal condition is 11000r/min in the step 3,12min.Other step and parameter are identical with embodiment one.
Embodiment six: this embodiment and embodiment one are different is to discard in the step 4 behind the 0.3ml supernatant on shaking table with 120r/min concussion 2min mixing.Other step and parameter are identical with embodiment one.
Embodiment seven: what this embodiment and embodiment one were different is that staining fluid is a methylene blue in the step 5.Other step and parameter are identical with embodiment one.
Embodiment eight: what this embodiment and embodiment one were different is to get 5ul bacterium drop in the step 5 to slide glass, and dry back is with the staining fluid 2min that dyes.Other step and parameter are identical with embodiment one.
Embodiment nine: the method for total plate count realizes according to the following steps in a kind of rapid detection solid food of this embodiment: one, under aseptic condition, accurately take by weighing the 25g solid sample; Put into 55ml concentration and be 0.5% peptone water solution, and on shaking table with 160r/min concussion 2min mixing; Two, draw 1ml sample centrifugal 8min under the condition of 1000r/min; Three, get 0.8ml supernatant recentrifuge 12min under the condition of 11000r/min; Four, discard behind the 0.3ml supernatant on shaking table with 120r/min concussion 2min mixing; Five, get 5ul bacterium drop to slide glass, dry back is with methylene blue staining fluid dyeing 2min; Six, dry with lens wiping paper behind the distilled water flushing; Seven, with calculating through the total plate count rapid detection system after the microscopic examination, can obtain total plate count.Other step and parameter are identical with embodiment one.
Claims (8)
1. the method for total plate count in the rapid detection solid food, it is characterized in that: the method for total plate count realizes according to the following steps in the rapid detection solid food: one, take by weighing the 20-30g solid sample under the aseptic condition and be dissolved in concussion mixing in back in the peptone water solution of 50-60ml; Two, absorption 1ml sample is centrifugal; Three, get 0.8ml supernatant recentrifuge; Four, discard mixing behind the 0.3ml supernatant; Five, get 5ul bacterium drop to slide glass, dry back is with the staining fluid 1-3min that dyes; Six, dry with lens wiping paper behind the distilled water flushing; Seven, with calculating through the total plate count rapid detection system after the microscopic examination, can obtain total plate count.
2. the method for total plate count in a kind of rapid detection solid food according to claim 1 is characterized in that in the step 1 adopting the peptone water solution of 0.5% concentration.
3. the method for total plate count in a kind of rapid detection solid food according to claim 1; It is characterized in that under aseptic condition, accurately taking by weighing in the step 1 25g solid sample; Put into the peptone water solution of 55ml, and on shaking table, shake the 2min mixing with 160r/min.
4. the method for total plate count in a kind of rapid detection solid food according to claim 1 is characterized in that centrifugal condition is 1000r/min in the step 2,8min.
5. the method for total plate count in a kind of rapid detection solid food according to claim 1 is characterized in that centrifugal condition is 11000r/min in the step 3,12min.
6. the method for total plate count in a kind of rapid detection solid food according to claim 1 is characterized in that discarding in the step 4 behind the 0.3ml supernatant on shaking table with 120r/min concussion 2min mixing.
7. the method for total plate count in a kind of rapid detection solid food according to claim 1 is characterized in that staining fluid is a methylene blue in the step 5.
8. the method for total plate count in a kind of rapid detection solid food according to claim 1 is characterized in that getting in the step 5 5ul bacterium drop to slide glass, and dry back is with the staining fluid 2min that dyes.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2010105275604A CN102453745A (en) | 2010-11-02 | 2010-11-02 | Method for quickly detecting total bacterial count in solid food |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2010105275604A CN102453745A (en) | 2010-11-02 | 2010-11-02 | Method for quickly detecting total bacterial count in solid food |
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| CN102453745A true CN102453745A (en) | 2012-05-16 |
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| CN2010105275604A Pending CN102453745A (en) | 2010-11-02 | 2010-11-02 | Method for quickly detecting total bacterial count in solid food |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103575580A (en) * | 2012-08-08 | 2014-02-12 | 内蒙古伊利实业集团股份有限公司 | Aseptic acquisition and inspection method for packing material |
| CN111044691A (en) * | 2019-12-24 | 2020-04-21 | 光明乳业股份有限公司 | Detection method of fungi in yoghourt |
-
2010
- 2010-11-02 CN CN2010105275604A patent/CN102453745A/en active Pending
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103575580A (en) * | 2012-08-08 | 2014-02-12 | 内蒙古伊利实业集团股份有限公司 | Aseptic acquisition and inspection method for packing material |
| CN103575580B (en) * | 2012-08-08 | 2016-02-17 | 内蒙古伊利实业集团股份有限公司 | The aseptic collection of packaging material and censorship method |
| CN111044691A (en) * | 2019-12-24 | 2020-04-21 | 光明乳业股份有限公司 | Detection method of fungi in yoghourt |
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Application publication date: 20120516 |