CN106896226A - A kind of method for quick of staphylococcus aureus - Google Patents
A kind of method for quick of staphylococcus aureus Download PDFInfo
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- CN106896226A CN106896226A CN201710294629.5A CN201710294629A CN106896226A CN 106896226 A CN106896226 A CN 106896226A CN 201710294629 A CN201710294629 A CN 201710294629A CN 106896226 A CN106896226 A CN 106896226A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
- G01N33/56911—Bacteria
- G01N33/56938—Staphylococcus
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/531—Production of immunochemical test materials
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/195—Assays involving biological materials from specific organisms or of a specific nature from bacteria
- G01N2333/305—Assays involving biological materials from specific organisms or of a specific nature from bacteria from Micrococcaceae (F)
- G01N2333/31—Assays involving biological materials from specific organisms or of a specific nature from bacteria from Micrococcaceae (F) from Staphylococcus (G)
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Abstract
The present invention provides a kind of method for quick of staphylococcus aureus, is related to the detection technique field of staphylococcus aureus.A kind of method for quick of staphylococcus aureus, comprises the following steps：1. culture medium is prepared；2. sterile working takes 25 g (mL) samples in homogenizing bag, and is added thereto to the enriched medium after 225mL preheatings, is put into the min of homogeneous 2 in homogenizer；42 DEG C of static or shaken cultivation 6h, can also extend to 24h as needed；The above-mentioned enrichment liquids of 0.1mL are taken, is added in the 5mL glass tubes containing 1mL selective mediums, 42 ± 1 DEG C are continued to cultivate 18 24h；3. culture is well mixed, takes 1mL to boiling water in test tube（100℃）Heating 10min, is cooled to room temperature, and 34 sample wells for dropping to colloidal-gold detecting-card are added dropwise with dropper, reacts 5 10 min, and sentence read result is invalid more than 30min colour developings.A kind of present invention conveniently and effectively approach for the fast qualitative detection of staphylococcus aureus is provided.
Description
Technical field
The present invention relates to the detection technique field of staphylococcus aureus, especially a kind of staphylococcus aureus is quick
Detection method.
Background technology
Staphylococcus aureus is a kind of important pathogen of the mankind, is under the jurisdiction of staphylococcus
（Staphylococcus）, there is the another name of " thermophilic meat bacterium ", it is the representative of gram-positive bacteria, many severe infections can be caused.Gold
Staphylococcus aureus are ubiquitous in nature, can all be found in the excreta of empty gas and water, dust and humans and animals.Therefore,
The chance that food is contaminated is a lot.The Center for Disease Control reports that the infection caused by staphylococcus aureus accounts for second
Position, is only second to Escherichia coli.Staphylococcus aureus enterotoxin is a worldwide health problem, in the U.S. by Staphylococcus aureus
The food poisoning that bacterium enterotoxin causes, accounts for the 33% of whole food posioning, Canadian then more, accounts for 45%, China's gold
The food poisoning that staphylococcus aureus cause also happens occasionally.
Detection means accurately and timely is the key for preventing and controlling pathogenic microorganism to propagate.Current staphylococcus aureus
Detection more than using traditional microculture method, mainly separated including preceding increasing bacterium, selective enrichment, flat board, biochemical examination
Test with the step such as serological Identification, the experiment that is related to is more, operate more loaded down with trivial details, detection cycle (generally requiring 4~7d) more long, accurate
Standby and round-off work is heavy, and the reality need of pathogenic bacteria quick detection in food at this stage cannot have been met.
The content of the invention
In order to solve the above-mentioned technical problem, the present invention provides a kind of method for quick of staphylococcus aureus.
A kind of method for quick of staphylococcus aureus, comprises the following steps：
1. culture medium is prepared
1）Enriched medium is prepared：26.5 g enriched medium are weighed, 42 ± 1 DEG C of L of deionized water 1 of preheating are added, filled
Divide dissolving, it is standby；
2）Selective medium is prepared：7.5 g Selective agar mediums are weighed, 42 ± 1 DEG C of deionized waters 100 of preheating are added
ML, fully dissolving, it is standby；
2. sterile working takes 25 g (mL) samples in homogenizing bag, and is added thereto to the Zengjing Granule after 225 mL preheatings
Base, is put into the min of homogeneous 2 in homogenizer；42 DEG C of static or h of shaken cultivation 6, can also extend to 24 as needed
h；The above-mentioned enrichment liquids of 0.1 mL are taken, is added in the 5 mL glass tubes containing 1 mL selective mediums, 42 ± 1 DEG C of continuation
Culture 18-24 h；
3. culture is well mixed, takes 1 mL to boiling water in test tube（100 ℃）10 min are heated, room temperature is cooled to, used
Dropper is added dropwise the sample well that 3-4 drops to colloidal-gold detecting-card, reacts 5-10 min, and sentence read result develops the color more than 30 min
It is invalid.
Further, the enriched medium includes following raw material（Count by weight）：12~14g of yeast extract, soybean
3~3.5g of peptone, 13~16g of agar, 17~19g of sodium chloride, 4~4.2g of Sodium Pyruvate .8~1 .1g of glycine 0, lithium chloride 0
.3~0 .5g, 4- methyl umbellate form ketone-β -5.5~6.2g of D-Glucose glycosides, to nitro β -0.2~0.5g of D-Glucose glycosides, to nitre
1.2~1.5g of base phenol-α-D- galactopyranoses.
Further, the Selective agar medium includes following raw material：Tryptone 25g；Glucose Liquid 200ml；Taro acid 5g；
The .5g of coetsoidin A 0；Citrate of magnesia 25g；8~12mg of Sodium Pyruvate；The .6g of potassium tellurite 0；The .2g of agar 0；Ambonic acid 0
.1g；200~the 300ml of potato water cooking liquid of concentration 300g/L；N- acetyl-D-glucose amine 5mg.
The beneficial effects of the invention are as follows：The invention provides the Staphylococcus aureus in a kind of detection sample of rapid sensitive
A kind of method of bacterium, for the fast qualitative detection of staphylococcus aureus provides conveniently and effectively approach.Shorten golden yellow Portugal
Grape coccus detection time, accelerates detection procedure, shortens the working time of testing staff, reduces labour intensity, saves water power and storage
The costs such as cleaning are deposited, detection efficiency is improved, testing cost is reduced, is easy to Site Detection, improve the degree of accuracy.
Specific embodiment
With reference to specific embodiment, the invention will be further described：
Embodiment one
A kind of method for quick of staphylococcus aureus, comprises the following steps：
1. culture medium is prepared
1）Enriched medium is prepared：26.5 g enriched medium are weighed, is added and is preheated 42 DEG C of L of deionized water 1, it is fully molten
Solution, it is standby；
2）Selective medium is prepared：7.5 g Selective agar mediums are weighed, 42 DEG C of mL of deionized water 100 of preheating are added, filled
Divide dissolving, it is standby；
2. sterile working takes 25 g (mL) samples in homogenizing bag, and is added thereto to the Zengjing Granule after 225 mL preheatings
Base, is put into the min of homogeneous 2 in homogenizer；42 DEG C of static or h of shaken cultivation 6, can also extend to 24 as needed
h；The above-mentioned enrichment liquids of 0.1 mL are taken, is added in the 5 mL glass tubes containing 1 mL selective mediums, 42 DEG C are continued to cultivate
21 h；
3. culture is well mixed, takes 1 mL to boiling water in test tube（100 ℃）10 min are heated, room temperature is cooled to, used
Dropper is added dropwise the sample well that 3-4 drops to colloidal-gold detecting-card, reacts 5-10 min, and sentence read result develops the color more than 30 min
It is invalid.
Wherein, the enriched medium includes following raw material（Count by weight）：Yeast extract 13g, soya peptone 3.3g, fine jade
Fat 15g, sodium chloride 18g, Sodium Pyruvate 4.1g, glycine 1g, lithium chloride 0.4g, 4- methyl umbellate form ketone-β-D-Glucose glycosides 6,
To nitro β-D-Glucose glycosides 0.3g, pnitrophenylα Dgalactospyranoside 1.4g.
Wherein, the Selective agar medium includes following raw material（Count by weight）：Tryptone 25g；Glucose Liquid
200ml；Taro acid 5g；The .5g of coetsoidin A 0；Citrate of magnesia 25g；Sodium Pyruvate 8mg；The .6g of potassium tellurite 0；The .2g of agar 0；
The .1g of ambonic acid 0；The potato water cooking liquid 200ml of concentration 300g/L；N- acetyl-D-glucose amine 5mg.
Embodiment two
A kind of method for quick of staphylococcus aureus, comprises the following steps：
1. culture medium is prepared
1）Enriched medium is prepared：26.5 g enriched medium are weighed, is added and is preheated 43 DEG C of L of deionized water 1, it is fully molten
Solution, it is standby；
2）Selective medium is prepared：7.5 g Selective agar mediums are weighed, 43 DEG C of mL of deionized water 100 of preheating are added, filled
Divide dissolving, it is standby；
2. sterile working takes 25 g (mL) samples in homogenizing bag, and is added thereto to the Zengjing Granule after 225 mL preheatings
Base, is put into the min of homogeneous 2 in homogenizer；42 DEG C of static or h of shaken cultivation 6, can also extend to 24 as needed
h；The above-mentioned enrichment liquids of 0.1 mL are taken, is added in the 5 mL glass tubes containing 1 mL selective mediums, 43 DEG C are continued to cultivate
18-24 h；
3. culture is well mixed, takes 1 mL to boiling water in test tube（100 ℃）10 min are heated, room temperature is cooled to, used
Dropper is added dropwise the sample well that 3-4 drops to colloidal-gold detecting-card, reacts 5-10 min, and sentence read result develops the color more than 30 min
It is invalid.
Wherein, the enriched medium includes following raw material（Count by weight）：Yeast extract 14g, soya peptone 3.5g, fine jade
Fat 16g, sodium chloride 19g, Sodium Pyruvate 4.2g, the .1g of glycine 1, lithium chloride 0 .5g, 4- methyl umbellate form ketone-β-D-Glucose
Glycosides 6.2g, to nitro β-D-Glucose glycosides 0.5g, pnitrophenylα Dgalactospyranoside 1.5g.
Wherein, the Selective agar medium includes following raw material（Count by weight）：Tryptone 25g；Glucose Liquid
200ml；Taro acid 5g；The .5g of coetsoidin A 0；Citrate of magnesia 25g；Sodium Pyruvate 12mg；The .6g of potassium tellurite 0；Agar 0
.2g；The .1g of ambonic acid 0；The potato water cooking liquid 300ml of concentration 300g/L；N- acetyl-D-glucose amine 5mg.
Embodiment three
A kind of method for quick of staphylococcus aureus, comprises the following steps：
1. culture medium is prepared
1）Enriched medium is prepared：26.5 g enriched medium are weighed, is added and is preheated 41 DEG C of L of deionized water 1, it is fully molten
Solution, it is standby；
2）Selective medium is prepared：7.5 g Selective agar mediums are weighed, 41 DEG C of mL of deionized water 100 of preheating are added, filled
Divide dissolving, it is standby；
2. sterile working takes 25 g (mL) samples in homogenizing bag, and is added thereto to the Zengjing Granule after 225 mL preheatings
Base, is put into the min of homogeneous 2 in homogenizer；42 DEG C of static or h of shaken cultivation 6, can also extend to 24 as needed
h；The above-mentioned enrichment liquids of 0.1 mL are taken, is added in the 5 mL glass tubes containing 1 mL selective mediums, 41 DEG C are continued to cultivate
18-24 h；
3. culture is well mixed, takes 1 mL to boiling water in test tube（100 ℃）10 min are heated, room temperature is cooled to, used
Dropper is added dropwise the sample well that 3-4 drops to colloidal-gold detecting-card, reacts 5-10 min, and sentence read result develops the color more than 30 min
It is invalid.
Wherein, the enriched medium includes following raw material（Count by weight）：Yeast extract 12g, soya peptone 3g, agar
13rd, sodium chloride 17g, Sodium Pyruvate 4g, the .8g of glycine 0, lithium chloride 0 .3g, 4- methyl umbellate form ketone-β-D-Glucose glycosides
5.5g, to nitro β-D-Glucose glycosides 0.2g, pnitrophenylα Dgalactospyranoside 1.2g.
Wherein, the Selective agar medium includes following raw material（Count by weight）：Tryptone 25g；Glucose Liquid
200ml；Taro acid 5g；The .5g of coetsoidin A 0；Citrate of magnesia 25g；Sodium Pyruvate 10mg；The .6g of potassium tellurite 0；Agar 0
.2g；The .1g of ambonic acid 0；The potato water cooking liquid 250ml of concentration 300g/L；N- acetyl-D-glucose amine 5mg.
Finally it should be noted that specific embodiment is unrestricted to of the invention further illustrating, to this area
For those of ordinary skill, can be done in the case where substance of the present invention is not departed from and further converted, and all these changes
Change the protection domain that should all belong to appended claims of the present invention.
Claims (3)
1. a kind of method for quick of staphylococcus aureus, it is characterised in that：
Comprise the following steps：
1. culture medium is prepared
1）Enriched medium is prepared：26.5 g enriched medium are weighed, 42 ± 1 DEG C of L of deionized water 1 of preheating are added, filled
Divide dissolving, it is standby；
2）Selective medium is prepared：7.5 g Selective agar mediums are weighed, 42 ± 1 DEG C of deionized waters 100 of preheating are added
ML, fully dissolving, it is standby；
2. sterile working takes 25 g (mL) samples in homogenizing bag, and is added thereto to the Zengjing Granule after 225 mL preheatings
Base, is put into the min of homogeneous 2 in homogenizer；42 DEG C of static or h of shaken cultivation 6, can also extend to 24 as needed
h；The above-mentioned enrichment liquids of 0.1 mL are taken, is added in the 5 mL glass tubes containing 1 mL selective mediums, 42 ± 1 DEG C of continuation
Culture 18-24 h；
3. culture is well mixed, takes 1 mL to boiling water in test tube（100 ℃）10 min are heated, room temperature is cooled to, used
Dropper is added dropwise the sample well that 3-4 drops to colloidal-gold detecting-card, reacts 5-10 min, and sentence read result develops the color more than 30 min
It is invalid.
2. the method for quick of a kind of staphylococcus aureus according to claim 1, it is characterised in that：Yeast extract 12
~14g, 3~3.5g of soya peptone, 13~16g of agar, 17~19g of sodium chloride, 4~4.2g of Sodium Pyruvate .8~1 of glycine 0
.1g .3~0 .5g, 4- methyl umbellate form ketone-β of lithium chloride 0-5.5~6.2g of D-Glucose glycosides, to nitro β-D-Glucose glycosides
0.2~0.5g, 1.2~1.5g of pnitrophenylα Dgalactospyranoside.
3. the method for quick of a kind of staphylococcus aureus according to claim 1, it is characterised in that：The selection
Culture medium includes following raw material：Tryptone 25g；Glucose Liquid 200ml；Taro acid 5g；The .5g of coetsoidin A 0；Citrate of magnesia
25g；8~12mg of Sodium Pyruvate；The .6g of potassium tellurite 0；The .2g of agar 0；The .1g of ambonic acid 0；The potato of concentration 300g/L
200~300ml of water cooking liquid；N- acetyl-D-glucose amine 5mg.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111961704A (en) * | 2020-07-22 | 2020-11-20 | 郑州中检科测试技术有限公司 | Quantitative detection method for staphylococcus aureus in food |
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| CN105603043A (en) * | 2016-04-08 | 2016-05-25 | 董建红 | Staphylococcus aureus selective culture medium |
| CN106282305A (en) * | 2016-11-02 | 2017-01-04 | 百奥森（江苏）食品安全科技有限公司 | A kind of detection method of Staphylococcus aureus in food |
| CN106370860A (en) * | 2016-08-24 | 2017-02-01 | 天津中新科炬生物制药有限公司 | Kit and test paper strip for serum immunoglobulin E colloidal gold chromatography quantitative detection |
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2017
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| CN105603043A (en) * | 2016-04-08 | 2016-05-25 | 董建红 | Staphylococcus aureus selective culture medium |
| CN106370860A (en) * | 2016-08-24 | 2017-02-01 | 天津中新科炬生物制药有限公司 | Kit and test paper strip for serum immunoglobulin E colloidal gold chromatography quantitative detection |
| CN106568951A (en) * | 2016-10-26 | 2017-04-19 | 北京农业质量标准与检测技术研究中心 | Nucleic acid aptamer-based escherichia coli O157:H7 colloidal gold test strip, and detection method |
| CN106282305A (en) * | 2016-11-02 | 2017-01-04 | 百奥森（江苏）食品安全科技有限公司 | A kind of detection method of Staphylococcus aureus in food |
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| 杜玉萍 等: "胶体金免疫层析法检测金黄色葡萄球菌的初步研究", 《热带医学杂志》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN111961704A (en) * | 2020-07-22 | 2020-11-20 | 郑州中检科测试技术有限公司 | Quantitative detection method for staphylococcus aureus in food |
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Application publication date: 20170627 |