CN104388529A - Fluorescence developing culture medium for detecting staphylococcus aureus - Google Patents
Fluorescence developing culture medium for detecting staphylococcus aureus Download PDFInfo
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- CN104388529A CN104388529A CN201410708100.XA CN201410708100A CN104388529A CN 104388529 A CN104388529 A CN 104388529A CN 201410708100 A CN201410708100 A CN 201410708100A CN 104388529 A CN104388529 A CN 104388529A
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Abstract
The invention discloses a fluorescence developing culture medium for detecting staphylococcus aureus. The fluorescence developing culture medium comprises a nitrogen source, a carbon source, sodium chloride and an infectious microbe growth inhibiter and also comprises a staphylococcus aureus developing substrate 4-methyl belliferyl-p-nitro beta-D-glucoside, beta-D-glucoside, a non-staphylococcus aureus developing substrate paranitrophenol-alpha-D-galactopyranose, and a non-staphylococcus aureus developing substrate paranitrophenol beta-D-glucuronide. Under the condition that a developing additive of the fluorescence developing culture medium is not available, a relatively simple and convenient developing scheme is provided to distinguish staphylococcus aureus from other staphylococci, the staphylococcus aureus can be effectively distinguished from a part of non-staphylococcus aureus and other infectious microbes, and relatively good selectivity is achieved for the staphylococcus aureus.
Description
Technical field
The present invention relates to a kind of substratum of microorganism detection, particularly relating to a kind of fluorescence developing substratum for detecting streptococcus aureus.
Background technology
Streptococcus aureus (Staphylococcus aureus) is conditioned pathogen, it is modal pathogenic bacteria in mankind's suppurative infection, also be a kind of important pathogen of the mankind, be under the jurisdiction of Staphylococcus (Staphylococcus), having the another name of " addicted to meat bacterium ", is the representative of gram-positive microorganism, can cause many local suppurative infection, also can cause pneumonia, pseudomembranous enteritis, pericarditis etc., even cause the systemic infection such as septicemia, Sepsis.When streptococcus aureus pollutes starch-containing and that moisture is more food, as milk box milk preparation, meat, egg etc., when temperature condition is suitable for, a considerable amount of enterotoxin can be produced through 8-10 hour, thus cause acute gastroenteritis symptom in various degree.
Streptococcus aureus is cultivated mainly through the colour developing of Barid-Parker agar selective medium.On this substratum, streptococcus aureus presents black colonies, has transparent circle around.But research finds, some non-streptococcus aureuses such as Staphylococcus saprophyticus, slowly staphylococcus and Sai Shi staphylococcus show above-mentioned feature equally, cause false positive.Patent of invention CN201310603799.9 discloses one for distinguishing streptococcus aureus and other staphylococcic color developing culture mediums, better selects chromatic effect to reach simultaneously, selects the specific developer that helps to help colour developing.
Summary of the invention
The object of the present invention is to provide a kind of fluorescence developing substratum detecting streptococcus aureus, helping in the non-existent situation of developer, in order to distinguish, streptococcus aureus and other are staphylococcic provides easier colour developing scheme.
Of the present invention adopted technical scheme is:
A kind of color developing culture medium for detecting streptococcus aureus, containing nitrogenous source, carbon source, sodium-chlor, varied bacteria growing inhibitor, also comprise streptococcus aureus chromogenic substrate 4-methyl umbellate form ketone-to nitro β-D-Glucose glycosides, β-D-Glucose glycosides, non-streptococcus aureus chromogenic substrate pnitrophenylα Dgalactospyranoside, non-streptococcus aureus chromogenic substrate p-NP β-D-Glucose aldehydic acid glycosides.
Preferably, described 4-methyl umbellate form ketone-β-D-Glucose glycosides is 0.5-5g/L.
Preferably, described 4-methyl umbellate form ketone-β-D-Glucose glycosides is 1-2g/L.
Preferably, described is 1-6g/L to nitro β-D-Glucose glycosides.
Preferably, described is 3g/L to nitro β-D-Glucose glycosides.
Preferably, the content of described nitrophenol-α-D-galactopyranose is 2-5g/L.
Preferably, described nitrophenols β-D-Glucose aldehydic acid glycosides is 0.6-1.5g/L.
Preferably, above-mentioned color developing culture medium, every 1000mL substratum contains extractum carnis 3-10g, peptone 5-15g, agar 10-30g, sodium-chlor 10-20g, bovine bile 1-5g, Cefixime Micronized 0.01-0.06mg, Sodium.alpha.-ketopropionate 5-15g.
Preferably, described substratum is also containing staphylococcus aureus growth promotor Sodium.alpha.-ketopropionate.
Extractum carnis, peptone, agar, sodium-chlor provide suitable growing environment for aureus growth, are beneficial to it and increase fast.High salt condition can promote staphylococcus aureus growth, suppresses varied bacteria growing simultaneously.
Sodium.alpha.-ketopropionate is the growth stimulant of streptococcus aureus.
Cefixime Micronized can suppress the growth of most of Gram-negative bacteria, and bovine bile can suppress the growth of most of gram-positive microorganism.
Nitrophenol-α-D-galactopyranose is the chromogenic substrate of α-d-galactosidase, can be degraded by non-streptococcus aureus, produces p-nitrophenol.
Nitrophenols β-D-Glucose aldehydic acid is the chromogenic substrate of β-D-Glucose aldehydic acid enzyme, can be degraded by non-streptococcus aureus, produces p-NP.
Streptococcus aureus and other staphylococcuses can act on 4-methyl umbellate form ketone-β-D-Glucose glycosides, generate the 4-methyl umbellate form ketone with intense fluorescence, visible colonies under generation fluorescent microscope.The alkaline phosphatase that streptococcus aureus and other staphylococcuses produce acts on nitro β-D-Glucose glycosides, generates p-nitrophenyl, produces yellow color colonies.But streptococcus aureus can not decompose nitrophenol-α-D-galactopyranose, nitrophenols β-D-Glucose aldehydic acid.And other staphylococcuses can decompose nitrophenol-α-D-galactopyranose or nitrophenols β-D-Glucose aldehydic acid, produce yellow product, thus produce the apparent colony colour the same with streptococcus aureus.
Therefore in the fluorescence developing substratum that the present invention announces, non-staphylococcus shows the color of bacterium colony itself, and staphylococcus display yellow color colonies color, streptococcus aureus display yellow color colonies color is also visible under fluorescent microscope.
The invention has the beneficial effects as follows: color developing culture medium of the present invention is that the detection of streptococcus aureus provides a kind of fluorescence developing scheme, also effectively can distinguish part staphylococcus and non-staphylococcus, streptococcus aureus and non-streptococcus aureus helping in the non-existent situation of developer, simplify bacterium colour developing culturing process.
Embodiment
A kind of color developing culture medium for detecting streptococcus aureus, containing nitrogenous source, carbon source, sodium-chlor, varied bacteria growing inhibitor, also comprise streptococcus aureus chromogenic substrate 4-methyl umbellate form ketone-to nitro β-D-Glucose glycosides, β-D-Glucose glycosides, non-streptococcus aureus chromogenic substrate pnitrophenylα Dgalactospyranoside, non-streptococcus aureus chromogenic substrate p-NP β-D-Glucose aldehydic acid glycosides.
The optional content range of 4-methyl umbellate form ketone-β-D-Glucose is 0.5-5g/L.
Be 1-6g/L to the optional content range of nitro β-D-Glucose glycosides.
The optional content range of the content of nitrophenol-α-D-galactopyranose is 2-5g/L.
Above-mentioned color developing culture medium, every 1000mL substratum contains extractum carnis 3-10g, peptone 5-15g, agar 10-30g, sodium-chlor 10-20g, bovine bile 1-5g, Cefixime Micronized 0.01-0.06mg, Sodium.alpha.-ketopropionate 5-15g.
Above-mentioned substratum is also containing staphylococcus aureus growth promotor Sodium.alpha.-ketopropionate.
Color developing culture medium of the present invention, for detecting the method for streptococcus aureus, comprises the steps:
(1) preparation colour developing is dull and stereotyped: add the raw material of above-mentioned color developing culture medium except nitrophenol-α-D-galactopyranose, nitrophenols β-D-Glucose aldehydic acid glycosides, Cefixime Micronized in every 1000mL deionized water, stir, heated and boiled is to dissolving completely, wait to be chilled to about 50 DEG C, add the nitrophenol-α-D-galactopyranose of filtration sterilization, nitrophenols β-D-Glucose aldehydic acid glycosides, Cefixime Micronized, mixing, is down flat plate, for subsequent use;
(2) inoculation culture: by sample or be inoculated on colour developing flat board containing the enrichment liquid of sample, 37 DEG C of overnight incubation are cultivated;
(3) possible colour developing result: do not develop the color under yellow fluorescence microscope, yellow and develop the color under fluorescent microscope, bacterium colony intrinsic colour.
Above-mentioned three kinds of color result are with distinct contrast, easily distinguish.
Below in conjunction with specific embodiment, the present invention is further illustrated, but do not limit to so.
Embodiment one
A kind of fluorescence developing substratum for detecting streptococcus aureus, every 1000mL substratum contains extractum carnis 3g, peptone 10g, agar 15g, sodium-chlor 10g, bovine bile 1.5g, Cefixime Micronized 0.04mg, 4-methyl umbellate form ketone-β-D-Glucose glycosides 2g/L is 3.5g/L to the content of nitro β-D-Glucose glycosides 3g/L, nitrophenol-α-D-galactopyranose, nitrophenols β-D-Glucose aldehydic acid glycosides is 1g/L, Sodium.alpha.-ketopropionate 10g.
Embodiment two
A kind of fluorescence developing substratum for detecting streptococcus aureus, every 1000mL substratum contains extractum carnis 3g, peptone 10g, agar 15g, sodium-chlor 10g, bovine bile 1.5g, Cefixime Micronized 0.04mg, 4-methyl umbellate form ketone-β-D-Glucose glycosides 3g/L is 2g/L to the content of nitro β-D-Glucose glycosides 2g/L, nitrophenol-α-D-galactopyranose, nitrophenols β-D-Glucose aldehydic acid glycosides is 1.5g/L, Sodium.alpha.-ketopropionate 8g.
Embodiment three
A kind of fluorescence developing substratum for detecting streptococcus aureus, every 1000mL substratum contains extractum carnis 3g, peptone 10g, agar 15g, sodium-chlor 15g, bovine bile 1.5g, Cefixime Micronized 0.04mg, 4-methyl umbellate form ketone-β-D-Glucose glycosides 5g/L, to nitro β-D-Glucose glycosides 6g/L, the content of nitrophenol-α-D-galactopyranose is 5g/L, and nitrophenols β-D-Glucose aldehydic acid glycosides is 0.6g/L, Sodium.alpha.-ketopropionate 5g.
Listed for table 17 kinds of reference cultures are made respectively the standard bacteria suspension (concentration is 107 ~ 108cfu/mL) of proper concn, streak inoculation is on the flat board made with embodiment 1 fluorescence developing substratum respectively, 37 DEG C of incubated overnight.Specificity experiments the results are shown in Table 1.In table 1 colony growth situation with growth a few district represent, 4th district represent well-grown, bacterium colony normal in size, 1-3 district represent growth suppressed, bacterium colony less than normal or growth be less than 4th district.
The specific chromogenic test-results of table 1 streptococcus aureus color developing culture medium
Experimental result shows, on the fluorescence developing substratum of Invention Announce, and streptococcus aureus displaing yellow and developing the color under fluorescent microscope, non-streptococcus aureus display bacterium colony intrinsic colour or only show yellow visible light color.
As can be seen here, the color developing culture medium that the present invention announces well can distinguish streptococcus aureus and non-streptococcus aureus, has better specificity to detection streptococcus aureus.
The above is only preferred embodiment of the present invention, and not do any pro forma restriction to the present invention, every any simple modification, equivalent variations done above embodiment according to technical spirit of the present invention, all falls into protection scope of the present invention.
Claims (9)
1. one kind for detecting the fluorescence developing substratum of streptococcus aureus, containing nitrogenous source, carbon source, sodium-chlor, varied bacteria growing inhibitor, it is characterized in that, also comprise streptococcus aureus chromogenic substrate 4-methyl umbellate form ketone-to nitro β-D-Glucose glycosides, β-D-Glucose glycosides, non-streptococcus aureus chromogenic substrate pnitrophenylα Dgalactospyranoside, non-streptococcus aureus chromogenic substrate p-NP β-D-Glucose aldehydic acid glycosides.
2. fluorescence developing substratum according to claim 1, is characterized in that, described 4-methyl umbellate form ketone-β-D-Glucose glycosides is 0.5-5g/L.
3. fluorescence developing substratum according to claim 2, is characterized in that, described 4-methyl umbellate form ketone-β-D-Glucose glycosides is 1-2g/L.
4. fluorescence developing substratum according to claim 1, is characterized in that, described is 1-6g/L to nitro β-D-Glucose glycosides.
5. fluorescence developing substratum according to claim 1, is characterized in that, described is 3g/L to nitro β-D-Glucose glycosides.
6. fluorescence developing substratum according to claim 1, is characterized in that, the content of described nitrophenol-α-D-galactopyranose is 2-5g/L.
7. fluorescence developing substratum according to claim 1, is characterized in that, described nitrophenols β-D-Glucose aldehydic acid glycosides is 0.6-1.5g/L.
8. the fluorescence developing substratum according to claim 1-7, is characterized in that, every 1000mL substratum contains extractum carnis 3-10g, peptone 5-15g, agar 10-30g, sodium-chlor 10-20g, bovine bile 1-5g, Cefixime Micronized 0.01-0.06mg, Sodium.alpha.-ketopropionate 5-15g.
9. fluorescence developing substratum according to claim 8, is characterized in that, also containing staphylococcus aureus growth promotor Sodium.alpha.-ketopropionate.
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Cited By (2)
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| 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 |
| CN110295502A (en) * | 2019-06-10 | 2019-10-01 | 安踏（中国）有限公司 | A kind of bacterium index plane preparation method for material, bacterium instruction fabric kimonos dress |
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| CN102177249A (en) * | 2008-10-08 | 2011-09-07 | 生物梅里埃公司 | Reaction medium for staphylococcus aureus bacteria |
| CN103228795A (en) * | 2010-11-24 | 2013-07-31 | 3M创新有限公司 | Methods and compositions to detect a biological activity |
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- 2014-11-27 CN CN201410708100.XA patent/CN104388529A/en active Pending
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| CN102177249A (en) * | 2008-10-08 | 2011-09-07 | 生物梅里埃公司 | Reaction medium for staphylococcus aureus bacteria |
| CN103228795A (en) * | 2010-11-24 | 2013-07-31 | 3M创新有限公司 | Methods and compositions to detect a biological activity |
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| 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 |
| CN110295502A (en) * | 2019-06-10 | 2019-10-01 | 安踏（中国）有限公司 | A kind of bacterium index plane preparation method for material, bacterium instruction fabric kimonos dress |
| CN110295502B (en) * | 2019-06-10 | 2021-11-26 | 安踏（中国）有限公司 | Preparation method of bacteria indicating fabric, bacteria indicating fabric and garment |
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