Rapid detection chromogenic culture medium for pseudomonas

Abstract

The invention belongs to the field of microbiological detection, and discloses a rapid detection chromogenic culture medium for pseudomonas, and the chromogenic detection culture medium comprises a chromogenic substrate, an endogenous enzyme promoter, a basic culture medium and a bacteriostatic agent. Specifically, the pseudomonas can react with a chromogenic substrate, so that a bacterial colony is blue-green, and other non-pseudomonas does not grow or weakly grow under the action of a bacteriostatic agent, or the chromogenic substrate different from the pseudomonas is utilized to display purple red or other colors which are inconsistent with the color of the pseudomonas bacteria, so that the pseudomonas bacteria can be distinguished, and the pseudomonas bacteria can be distinguished. And rapid separation and identification are realized. According to the rapid detection chromogenic culture medium for the pseudomonas, the glycoside chromogenic substrate and the endogenous enzyme accelerant are utilized, the problem that when the rapid detection chromogenic culture medium for the pseudomonas is prepared, the pseudomonas has chromatic aberration or does not have color development is solved, and the blank of the rapid detection chromogenic culture medium for the pseudomonas in the domestic market is filled.

Description

Technical Field

[0001] The invention belongs to the field of microbial detection, and in particular relates to a color development culture medium for rapid detection of Pseudomonas. Background Art

[0002] Pseudomonas is a type of Gram-negative aerobic rod with low nutritional requirements, so it is widely found in animals, plants, fresh water, soil and natural environment. There are many species in this genus, among which Pseudomonas aeruginosa, Pseudomonas fluorescens, Pseudomonas putida and other conditional pathogens that humans focus on preventing and controlling. These bacteria can produce a variety of endotoxins and exotoxins, which can easily cause low immunity or cause different degrees of zoonotic diseases. They are key monitoring microorganisms for hospital infections. In addition, Pseudomonas bacteria are closely related to water sources, food processing, cosmetic preparation, and pharmaceutical engineering pollution. Therefore, they are also the main microorganisms for microbial contamination and control in the production process of drinking water industry, food industry, cosmetic industry, pharmaceutical industry and other fields.

[0003] At present, due to the lack of Pseudomonas-specific isolation culture media, microbiologists usually use pure culture media such as tryptic soy agar (TSA) to isolate contaminating microorganisms, purify them, and then identify them through various identification systems. The whole process takes at least 3-5 days. In addition, when there are many types of contaminating bacteria in the production process and they are very complex, it is usually difficult to separate them from pure culture media. For example, meat products often contain a large number of Proteus, which will migrate and grow on TSA culture media, covering Pseudomonas bacteria, making it difficult to isolate Pseudomonas, causing deviations in the formulation of microbial contamination control measures by enterprises, and still failing to prevent Pseudomonas contamination, resulting in huge economic losses.

[0004] Chromogenic culture medium is a rapid screening culture medium that combines the separation and identification of microorganisms by using specific enzyme substrates. Its advantages of saving time and effort, simple operation, high sensitivity, and good specificity make chromogenic culture medium very popular among people involved in microbial research and testing. At present, there are no chromogenic culture medium related products for rapid detection of Pseudomonas in the domestic market. The foreign company Chromagar produces this product, but has not announced its product technical plan. In addition, foreign products are expensive and have a long delivery time, which limits their promotion and application. Therefore, it is particularly important to develop a chromogenic culture medium with good specificity, high accuracy, and the ability to quickly separate Pseudomonas. Summary of the invention

[0005] The object of the present invention is to overcome at least one disadvantage of the prior art and provide a chromogenic culture medium for rapid detection of Pseudomonas.

[0006] The technical solution adopted by the present invention is: The first aspect of the present invention provides: A chromogenic culture medium for rapid detection of Pseudomonas, comprising a chromogenic substrate, an endogenous enzyme promoter, a basic culture medium and an antibacterial agent; Wherein, the endogenous enzyme promoter is selected from at least one of n-octyl-β-D-pyranoglucoside and n-octyl-β-D-lactoside; The color developing substrate is selected from at least one of 5-bromo-4-chloro-3-indolephosphocholine, 5-bromo-6-chloro-3-indole-β-D-galactoside, 5-bromo-6-chloro-3-indole-β-D-pyranoglucopyranoside and isopropyl-β-D-thiogalactoside.

[0007] In some examples of the rapid detection chromogenic culture medium for Pseudomonas, the total amount of the endogenous enzyme promoter in the culture medium is 3-12 g / L.

[0008] In some examples of Pseudomonas rapid detection chromogenic culture medium, the mass ratio of n-octyl-β-D-pyranoglucoside to n-octyl-β-D-lactosidoside is (5-10):(0.5-2).

[0009] In some examples of Pseudomonas rapid detection chromogenic culture medium, the total dosage of the endogenous enzyme promoter in the culture medium is 3-12 g / L, and the mass dosage ratio of n-octyl-β-D-pyranoglucoside and n-octyl-β-D-lactosidoside is (5-10): (0.5-2).

[0010] In some examples of chromogenic culture medium for rapid detection of Pseudomonas, the total amount of the chromogenic substrate in the culture medium is 0.1-1 g / L.

[0011] In some examples of chromogenic culture medium for rapid detection of Pseudomonas, the amount of 5-bromo-4-chloro-3-indolylphosphocholine used in the culture medium is 0.01-1.0 g / L.

[0012] In some examples of chromogenic culture medium for rapid detection of Pseudomonas, the amount of 5-bromo-6-chloro-3-indole-β-D-galactoside used in the culture medium is 0.01-0.5 g / L.

[0013] In some examples of chromogenic culture medium for rapid detection of Pseudomonas, the amount of isopropyl-β-D-thiogalactoside used in the culture medium is 0.01-0.5 g / L.

[0014] In some examples of rapid detection chromogenic culture medium for Pseudomonas, the amount of 5-bromo-6-chloro-3-indole-β-D-pyranoglucoside used in the culture medium is 0.01 to 1 g / L.

[0015] In some examples of chromogenic culture medium for rapid detection of Pseudomonas, the amount of the chromogenic substrate in the culture medium is: 5-bromo-4-chloro-3-indolephosphocholine 0.01-1.0 g / L, 5-bromo-6-chloro-3-indole-β-D-galactoside 0.01-0.5 g / L, isopropyl-β-D-thiogalactoside 0.01-0.5 g / L, and 5-bromo-6-chloro-3-indole-β-D-pyranoglucoside 0.01-1 g / L.

[0016] In some examples of the rapid detection chromogenic culture medium for Pseudomonas, the antibacterial agent includes at least one of nalidixic acid, vancomycin, sodium fusidate, cephalosporin II, novobiocin, and triclosan.

[0017] In some examples of the chromogenic culture medium for rapid detection of Pseudomonas, the total amount of the antibacterial agent in the culture medium is 0.025-0.18 g / L.

[0018] In some examples of chromogenic culture medium for rapid detection of Pseudomonas, the amount of sodium fusidate used in the culture medium is 0.01-0.05 g / L.

[0019] In some examples of chromogenic culture medium for rapid detection of Pseudomonas, the amount of nalidixic acid used in the culture medium is 0.01-0.1 g / L.

[0020] In some examples of rapid chromogenic culture media for Pseudomonas detection, the amount of vancomycin used in the culture medium is 0.005-0.03 g / L.

[0021] In some examples of Pseudomonas rapid detection chromogenic culture medium, the dosage of the antibacterial agent in the culture medium is: sodium fusidate 0.01-0.05 g / L, nalidixic acid 0.01-0.1 g / L, and vancomycin 0.005-0.03 g / L.

[0022] In some examples of the chromogenic medium for rapid detection of Pseudomonas, the basal medium comprises a carbon source, a nitrogen source, an inorganic salt, a gelling agent and water. Preferably, the gelling agent is agar.

[0023] In some examples of Pseudomonas rapid detection chromogenic culture medium, the composition of the basic culture medium is: 5.0-20.0 g / L peptone, 3.0-5.0 g / L trypticase, 3.0-5.0 g / L soytone, 4.0-8.0 g / L yeast powder, 5.0-10.0 g / L sodium chloride, 1.0-5.0 g / L magnesium chloride, and 12.0-15.0 g / L agar.

[0024] The above features can be combined arbitrarily unless they conflict with each other.

[0025] The second aspect of the present invention provides: A method for detecting Pseudomonas comprises culturing the bacteria to be detected on the Pseudomonas rapid detection color development medium described in the first aspect of the present invention, and observing the color development during the culturing process to determine whether Pseudomonas is present.

[0026] Specifically, Pseudomonas can react with chromogenic substrates to make the colonies appear blue-green, while other non-Pseudomonas bacteria do not grow or grow weakly under the action of antibacterial agents, or use chromogenic substrates different from those of Pseudomonas to appear purple-red or other colors that are inconsistent with the color of Pseudomonas bacteria, so as to facilitate the distinction between Pseudomonas bacteria and achieve rapid separation and identification.

[0027] The beneficial effects of the present invention are: 1. The chromogenic culture medium for rapid detection of Pseudomonas of the present invention fills the gap of chromogenic culture medium for rapid detection of Pseudomonas in the domestic market.

[0028] 2. The Pseudomonas rapid detection chromogenic culture medium of the present invention utilizes glycoside chromogenic substrates and endogenous enzyme promoters, thereby solving the problem of poor or no color development of Pseudomonas bacteria when preparing a Pseudomonas rapid detection chromogenic culture medium.

[0029] 3. The Pseudomonas rapid detection chromogenic culture medium of the present invention can effectively inhibit the growth of miscellaneous bacteria by optimizing the composition of the antibacterial agent, provides a technical solution to avoid interference from other bacteria, and can quickly achieve the rapid separation of Pseudomonas. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] Figure 1 The color development effect of the flat panel of Example 1 and Comparative Example 1 is compared.

[0031] Figure 2 Comparison of color rendering effects between Example 1 (left) and foreign products (right). DETAILED DESCRIPTION

[0032] A chromogenic culture medium for rapid detection of Pseudomonas, comprising a chromogenic substrate, an endogenous enzyme promoter, a basic culture medium and an antibacterial agent; Wherein, the endogenous enzyme promoter is selected from at least one of n-octyl-β-D-pyranoglucoside and n-octyl-β-D-lactoside; The color developing substrate is selected from at least one of 5-bromo-4-chloro-3-indolephosphocholine, 5-bromo-6-chloro-3-indole-β-D-galactoside, 5-bromo-6-chloro-3-indole-β-D-pyranoglucopyranoside and isopropyl-β-D-thiogalactoside.

[0033] The basic culture medium provides basic growth conditions for Pseudomonas, including carbon source, nitrogen source, inorganic salt, gelling agent and water. The function of the gelling agent is to form a plate, which is convenient for observing the color development of the colony. Preferably, the gelling agent is agar. The typical formula of the basic culture medium for Pseudomonas is: peptone 5.0-20.0 g / L, trypticase 3.0-5.0 g / L, soytone 3.0-5.0 g / L, yeast powder 4.0-8.0 g / L, sodium chloride 5.0-10.0 g / L, magnesium chloride 1.0-5.0 g / L, agar 12.0-15.0 g / L.

[0034] In some examples of the rapid detection chromogenic culture medium for Pseudomonas, the total amount of the endogenous enzyme promoter in the culture medium is 3-12 g / L.

[0035] Preferably, the endogenous enzyme promoter comprises a mixture of n-octyl-β-D-pyranoglucoside and n-octyl-β-D-lactoside. In some examples of rapid detection chromogenic medium for Pseudomonas, the mass ratio of n-octyl-β-D-pyranoglucoside to n-octyl-β-D-lactoside is (5-10): (0.5-2).

[0036] In some examples of the colorimetric culture medium for rapid detection of Pseudomonas, the total amount of the endogenous enzyme promoter in the culture medium is 3 to 12 g / L, and the mass ratio of n-octyl-β-D-pyranoglucoside to n-octyl-β-D-lactoside is (5 to 10): (0.5 to 2). In particular, each 1000 ml of the culture medium contains 5 to 10 g of n-octyl-β-D-pyranoglucoside and 0.5 to 2 g of n-octyl-β-D-lactoside.

[0037] In some examples of chromogenic culture medium for rapid detection of Pseudomonas, the total amount of the chromogenic substrate in the culture medium is 0.1-1 g / L.

[0038] In some examples of chromogenic culture medium for rapid detection of Pseudomonas, the amount of 5-bromo-4-chloro-3-indolylphosphocholine used in the culture medium is 0.01-1.0 g / L.

[0039] In some examples of chromogenic culture medium for rapid detection of Pseudomonas, the amount of 5-bromo-6-chloro-3-indole-β-D-galactoside used in the culture medium is 0.01-0.5 g / L.

[0040] In some examples of chromogenic culture medium for rapid detection of Pseudomonas, the amount of isopropyl-β-D-thiogalactoside used in the culture medium is 0.01-0.5 g / L.

[0041] In some examples of rapid detection chromogenic culture medium for Pseudomonas, the amount of 5-bromo-6-chloro-3-indole-β-D-pyranoglucoside used in the culture medium is 0.01 to 1 g / L.

[0042] Preferably, the chromogenic substrate is a mixture of 5-bromo-4-chloro-3-indolephosphocholine, 5-bromo-6-chloro-3-indole-β-D-galactoside, isopropyl-β-D-thiogalactoside and 5-bromo-6-chloro-3-indole-β-D-pyranoglucoside. In particular, each 1000 ml of the culture medium contains 0.01-1.0 g of 5-bromo-4-chloro-3-indolephosphocholine, 0.01-0.5 g of 5-bromo-6-chloro-3-indole-β-D-galactoside, 0.01-0.5 g of isopropyl-β-D-thiogalactoside and 0.01-1 g of 5-bromo-6-chloro-3-indole-β-D-pyranoglucoside.

[0043] The function of the antibacterial agent is to inhibit the growth of miscellaneous bacteria, so that Pseudomonas and miscellaneous bacteria can be better separated during the culture process. In some examples of Pseudomonas rapid detection color development medium, the antibacterial agent includes at least one of nalidixic acid, vancomycin, sodium fusidate, vancomycin II, novobiocin, and triclosan. The specific amount can be adjusted accordingly according to the antibacterial situation.

[0044] In some examples of the chromogenic culture medium for rapid detection of Pseudomonas, the total amount of the antibacterial agent in the culture medium is 0.025-0.18 g / L.

[0045] In some examples of chromogenic culture medium for rapid detection of Pseudomonas, the amount of sodium fusidate used in the culture medium is 0.01-0.05 g / L.

[0046] In some examples of chromogenic culture medium for rapid detection of Pseudomonas, the amount of nalidixic acid used in the culture medium is 0.01-0.1 g / L.

[0047] In some examples of rapid chromogenic culture media for Pseudomonas detection, the amount of vancomycin used in the culture medium is 0.005-0.03 g / L.

[0048] In some examples of Pseudomonas rapid detection chromogenic culture medium, the dosage of the antibacterial agent in the culture medium is 0.01-0.05 g / L sodium fusidate, 0.01-0.1 g / L nalidixic acid, and 0.005-0.03 g / L vancomycin.

[0049] Preferably, the antibacterial agent is a mixture of sodium fusidate, vancomycin and nalidixic acid. In particular, each 1000 ml of the culture medium contains 0.01-0.05 g sodium fusidate, 0.01-0.1 g nalidixic acid and 0.005-0.03 g vancomycin.

[0050] The above features can be combined arbitrarily unless they conflict with each other.

[0051] The following disclosure provides many different embodiments or examples for implementing different solutions of the present invention.

[0052] Example 1 A chromogenic culture medium for rapid detection of Pseudomonas, comprising a basic culture medium, a chromogenic substrate, an endogenous enzyme promoter and an antibacterial agent, wherein: Basic culture medium, each 1000 ml culture medium contains 10.0 g of peptone, 3.0 g of trypticase, 3.0 g of soytone, 4.0 g of yeast powder, 5.0 g of sodium chloride, 1.0 g of magnesium chloride, and 12.0 g of agar; Colorimetric substrate: Each 1000 ml culture medium contains 0.3 g of 5-bromo-6-chloro-3-indolephosphocholine, 0.2 g of 5-bromo-6-chloro-3-indole-β-D-galactoside, 0.1 g of isopropyl-β-D-thiogalactoside, and 0.2 g of 5-bromo-6-chloro-3-indole-β-D-pyranoglucopyranoside; Endogenous enzyme promoter: 6.0 g of n-octyl-β-D-pyranoglucoside and 0.5 g of n-octyl-β-D-lactoside per 1000 ml of culture medium; Antibacterial agent: Each 1000ml culture medium contains 0.03g sodium fusidate, 0.02g nalidixic acid, and 0.01g vancomycin.

[0053] Example 2 A chromogenic culture medium for rapid detection of Pseudomonas, comprising a basic culture medium, a chromogenic substrate, an endogenous enzyme promoter and an antibacterial agent, wherein: Basic culture medium, each 1000 ml culture medium contains 7.0 g of peptone, 5.0 g of trypticase, 5.0 g of soytone, 6.0 g of yeast powder, 5.0 g of sodium chloride, 2.0 g of magnesium chloride, and 15.0 g of agar; Colorimetric substrate: Each 1000 ml culture medium contains 0.1 g of 5-bromo-6-chloro-3-indolephosphocholine, 0.3 g of 5-bromo-6-chloro-3-indole-β-D-galactoside, and 0.2 g of isopropyl-β-D-thiogalactoside; Endogenous enzyme promoter: 8.0 g of n-octyl-β-D-pyranoglucoside and 0.5 g of n-octyl-β-D-lactoside per 1000 ml of culture medium; Antibacterial agent: Each 1000ml culture medium contains 0.02g of nalidixic acid and 0.1g of cephalosporin II.

[0054] Example 3 A chromogenic culture medium for rapid detection of Pseudomonas, comprising a basic culture medium, a chromogenic substrate, an endogenous enzyme promoter and an antibacterial agent, wherein: Basic culture medium, each 1000 ml culture medium contains 5.0 g of peptone, 9.0 g of trypticase, 5.0 g of soytone, 7.0 g of yeast powder, 6.0 g of sodium chloride, 1.0 g of magnesium chloride, and 15.0 g of agar; Colorimetric substrate: Each 1000 ml culture medium contains 0.2 g of 5-bromo-6-chloro-3-indolephosphocholine, 0.1 g of 5-bromo-6-chloro-3-indole-β-D-galactoside, 0.1 g of isopropyl-β-D-thiogalactoside, and 0.2 g of 5-bromo-6-chloro-3-indole-β-D-pyranoglucopyranoside; Endogenous enzyme promoter: Each 1000 ml culture medium contains 7.0 g of n-octyl-β-D-pyranoglucoside and 1.0 g of n-octyl-β-D-lactoside; Antibacterial agent: Each 1000ml culture medium contains nalidixic acid 0.01g, sodium fusidate 0.02g, and novobiocin 0.001g.

[0055] Comparative Example 1 Compared with Example 1, no endogenous enzyme promoter is contained, specifically: A Pseudomonas detection culture medium, comprising a basic culture medium, a chromogenic substrate and an antibacterial agent, wherein: Basic culture medium, each 1000 ml culture medium contains 10.0 g of peptone, 3.0 g of trypticase, 3.0 g of soytone, 4.0 g of yeast powder, 5.0 g of sodium chloride, 1.0 g of magnesium chloride, and 12.0 g of agar; Colorimetric substrate: Each 1000 ml culture medium contains 0.3 g of 5-bromo-6-chloro-3-indolephosphocholine, 0.2 g of 5-bromo-6-chloro-3-indole-β-D-galactoside, 0.1 g of isopropyl-β-D-thiogalactoside, and 0.2 g of 5-bromo-6-chloro-3-indole-β-D-pyranoglucopyranoside; Antibacterial agent: Each 1000ml culture medium contains 0.03g sodium fusidate, 0.02g nalidixic acid, and 0.01g vancomycin.

[0056] Comparative Example 2 Compared with Example 1, no antibacterial agent is contained, specifically: A Pseudomonas detection culture medium, comprising a basic culture medium, a chromogenic substrate and an endogenous enzyme promoter, wherein: Basic culture medium, each 1000 ml culture medium contains 10.0 g of peptone, 3.0 g of trypticase, 3.0 g of soytone, 4.0 g of yeast powder, 5.0 g of sodium chloride, 1.0 g of magnesium chloride, and 12.0 g of agar; Colorimetric substrate: Each 1000 ml culture medium contains 0.3 g of 5-bromo-6-chloro-3-indolephosphocholine, 0.2 g of 5-bromo-6-chloro-3-indole-β-D-galactoside, 0.1 g of isopropyl-β-D-thiogalactoside, and 0.2 g of 5-bromo-6-chloro-3-indole-β-D-pyranoglucopyranoside; Endogenous enzyme promoter: Each 1000 ml culture medium contains 6.0 g of n-octyl-β-D-pyranoglucoside and 0.5 g of n-octyl-β-D-lactosyltransferase.

[0057] Tablet performance test 1 Materials 1.1 Strains Pseudomonas aeruginosa ( Pseudomonas aeruginosa )ATCC27853, Pseudomonas aeruginosa ( Pseudomonas aeruginosa )CMCC(B)10104, Pseudomonas aeruginosa ( Pseudomonas aeruginosa )ATCC9027, Pseudomonas aeruginosa ( Pseudomonas aeruginosa )ATCC15442, Pseudomonas aeruginosa ( Pseudomonas aeruginosa )CMCC(B)10901, Pseudomonas fluorescens ( Pseudomonas fluorescens )GIM1.49, Pseudomonas putida ( Pseudomonas putida )GIM1.57, Pseudomonas alcaligenes ( Pseudomonas pseudoalcaligenes ) CGMCC1.1805, Pseudomonas stutzeri ( Pseudomonas stutzeri ) CGMCC1.1803, Pseudomonas mendocinae, Enterococcus faecalis ( Enterococcus faecalis )ATCC29212, Escherichia coli ( Escherichia coli )ATCC25922, Escherichia coli ( Escherichia coli )CMCC(B)43201, Escherichia coli ( Escherichia coli )CMCC(B)44102, Enterobacter aerogenes ( Enterobacter aerogenes )ATCC13048, Cronobacter sakazakii ( Enterobacter sakazakii )ATCC 29544, Salmonella typhimurium ( Salmonella typhimurium )ATCC 14028, Salmonella paratyphi B ( Salmonella paratyphi B )CMCC(B)50094, Klebsiella pneumoniae ( Klebsiella pneumoniae )CMCC(B)46117, Citrobacter freundii ( Citrobacter freundii )ATCC 43864, Enterobacter cloacae ( Enterobacter cloacae)CMCC(B)45301, Staphylococcus aureus ( Staphylococcus aureus )CMCC(B)26003, Proteus vulgaris ( Proteus vulgaris )CMCC(B)49027, Proteus mirabilis ( Proteus mirabilis )CMCC(B)49005 is kept by this laboratory.

[0058] 1.2 Culture medium and reagents The culture medium materials described in the examples are all common raw materials produced by Guangdong Huankai Microbiological Technology Co., Ltd., Huankai Trypticase Soy Agar Plate (TSA Plate for short), and CHROMagar Pseudomonas Chromogenic Medium.

[0059] 2 Methods 2.1 Evaluation of biological performance of culture medium Growth rate test: quantitative method is used. Prepare a fresh slant culture of target bacteria such as Pseudomonas aeruginosa for about 10 3 cfu / ml bacterial suspension. Take 0.1ml of each of the above bacterial solutions and inoculate them on the plates described in the above examples using a fully automatic spiral inoculator, and at the same time inoculate the reference culture medium TSA plates, and culture at 37±1°C for 18-24 hours. Perform 3 replicates on each plate. Observe the growth of each strain on the culture medium and count its colonies. Calculate the growth rate PR according to the following formula: PR=NS / N0, NS: the average number of colonies obtained on the test culture medium plate; N0: the average number of colonies obtained on the reference culture medium plate.

[0060] Selective test: non-target bacteria are made into 10 8 cfu / mL~10 9 cfu / mL concentration of bacterial suspension, use 1μL inoculation loop to take 1 loop of bacterial suspension, draw six parallel straight lines on the surface of the test medium and reference medium plate, and culture at 37±1℃ for 18-24 hours. After culture, calculate the growth index G of the culture medium according to the following method. Each line with relatively dense colony growth has a G of 1, and the maximum score on each culture dish is 6. If only half of the lines have colony growth, G is 0.5. If there is no colony growth on the line or the growth is less than half of the line, G is 0. Record the sum of the scores of each plate to get the G value.

[0061] 3 Results 3.1 Comparison of flat panel color rendering effects The culture medium described in Example 1 was compared with Comparative Example 1. The biological properties are shown in Table 1, wherein the color development effect of Pseudomonas aeruginosa ATCC27853 is as follows: Figure 1 shown.

[0062] Table 1. Comparison of color development effects of plates containing endogenous enzyme promoters and plates without endogenous enzyme promoters

[0063] Note: “-” indicates no color or the color of the colony itself; “+” indicates light color; “++” indicates medium intensity color; “+++” indicates strong color.

[0064] From Table 1 and Figure 1 It can be seen that compared with the plate without endogenous enzyme promoter, the color development culture medium of the present invention has a significant promoting effect on the coloring effect of target bacteria, has a high colony recognition, and can achieve rapid separation and identification of target bacteria.

[0065] 3.2 Comparison of biological effects of plates The culture medium described in Example 1 was compared with Comparative Example 2, and the biological properties are shown in Table 2: Table 2. Comparison of biological effects of Example 1 and Comparative Example 2

[0066] As shown in Table 2, compared with the plate without selective antibacterial agent, the color development medium of the present invention can effectively avoid the interference of Gram-negative bacteria, Gram-positive bacteria and some common bacteria on the detection of target bacteria except Pseudomonas, and realize the rapid separation and identification of target bacteria.

[0067] 3.3 Specificity test The culture medium described in Example 1 was compared with a foreign product (CHROMagar™ Pseudomonas). The biological properties are shown in Table 3. The color development effects of some strains are shown in Table 3. Figure 2 shown.

[0068] Table 3. Comparison between the tablet of Example 1 and foreign products

[0069] Note: “+” indicates light color; “++” indicates medium intensity color; “+++” indicates strong color.

[0070] From Table 3, Figure 2 It can be seen that the target bacteria growth effect of the color development medium described in the present invention is better than that of CHROMagar. In addition to meeting the conventional separation and identification of Pseudomonas aeruginosa, the separation effect of other Pseudomonas of the genus Pseudomonas (fluorescing Pseudomonas, Pseudomonas putida, etc.) is also better than CHROMagar, which greatly improves the application range of the product. The coloring effect of the target bacteria is also better than CHROMagar overall, which has the advantage of fast color development speed, greatly improving the inspection efficiency.

[0071] The results showed that the chromogenic culture medium for rapid detection of Pseudomonas described in the implementation case had obvious target bacteria specificity, typical colony color characteristics, and a strong selective inhibitory effect on non-target bacteria.

[0072] In summary, the chromogenic culture medium for rapid detection of Pseudomonas described in the present invention has the advantages of good color development effect, high specificity, simple result judgment, etc.

[0073] The above is a further detailed description of the present invention, which should not be regarded as a limitation on the specific implementation of the present invention. For ordinary technicians in the technical field to which the present invention belongs, simple deduction or replacement without departing from the concept of the present invention is within the protection scope of the present invention.

Claims

1. A chromogenic medium for rapid detection of Pseudomonas, characterized in that: A color development substrate, an endogenous enzyme promoter, a basic culture medium and an antibacterial agent are added; Wherein, the endogenous enzyme promoter is selected from at least one of n-octyl-β-D-pyranoglucoside and n-octyl-β-D-lactoside; The color developing substrate is selected from at least one of 5-bromo-4-chloro-3-indolephosphocholine, 5-bromo-6-chloro-3-indole-β-D-galactoside, 5-bromo-6-chloro-3-indole-β-D-pyranoglucopyranoside and isopropyl-β-D-thiogalactoside.

2. The chromogenic medium for rapid detection of Pseudomonas according to claim 1, characterized in that The total dosage of the endogenous enzyme promoter in the culture medium is 3-12 g / L.

3. The chromogenic medium for rapid detection of Pseudomonas according to claim 1 or 2, characterized in that: The mass usage ratio of n-octyl-β-D-pyranoglucoside and n-octyl-β-D-lactosidoside is (5-10):(0.5-2).

4. The chromogenic medium for rapid detection of Pseudomonas according to claim 1, characterized in that The total dosage of the color developing substrate in the culture medium is 0.1-1 g / L.

5. The chromogenic medium for rapid detection of Pseudomonas according to claim 1 or 4, characterized in that: The dosage of the color developing substrate in the culture medium is: 0.01-1.0 g / L of 5-bromo-4-chloro-3-indolephosphocholine, 0.01-0.5 g / L of 5-bromo-6-chloro-3-indole-β-D-galactoside, 0.01-0.5 g / L of isopropyl-β-D-thiogalactoside, and 0.01-1 g / L of 5-bromo-6-chloro-3-indole-β-D-pyranoglucoside.

6. The chromogenic medium for rapid detection of Pseudomonas according to claim 1, characterized in that: The antibacterial agent includes at least one of nalidixic acid, vancomycin, sodium fusidate, cephalosporin II, novobiocin and triclosan.

7. The chromogenic medium for rapid detection of Pseudomonas according to claim 1, characterized in that: The total dosage of the antibacterial agent in the culture medium is 0.025-0.18 g / L.

8. The chromogenic medium for rapid detection of Pseudomonas according to claim 6 or 7, characterized in that: The dosage of the antibacterial agent in the culture medium is: 0.01-0.05 g / L sodium fusidate, 0.01-0.1 g / L nalidixic acid, and 0.005-0.03 g / L vancomycin.

9. The chromogenic medium for rapid detection of Pseudomonas according to claim 1, characterized in that: The basic culture medium is composed of: 5.0-20.0 g / L of peptone, 3.0-5.0 g / L of trypticase, 3.0-5.0 g / L of soytone, 4.0-8.0 g / L of yeast powder, 5.0-10.0 g / L of sodium chloride, 1.0-5.0 g / L of magnesium chloride, and 12.0-15.0 g / L of agar.

10. A method for detecting Pseudomonas, characterized in that: The method comprises culturing the bacteria to be tested in the Pseudomonas rapid detection color development medium according to any one of claims 1 to 9, and observing the color development during the culturing process to determine whether Pseudomonas exists.

Images