CN110607243A - Culture medium for separating fungi and yeast-like fungi and preparation method thereof - Google Patents
Culture medium for separating fungi and yeast-like fungi and preparation method thereof Download PDFInfo
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Abstract
The invention relates to the technical field of medical chemiluminescence immunoassay detection, in particular to a culture medium for separating fungi and yeast-like fungi and a preparation method thereof, wherein tryptone, animal peptone and glucose are used for providing proper carbon source and nitrogen source for microorganisms, the proportion of the whole carbon source and nitrogen source is controllable, and the content of glucose is more, so that the pH of the whole culture medium tends to 5.5-6.5, which is beneficial to the growth of the fungi and the yeast-like fungi. Meanwhile, single chloramphenicol is used as a bacteria inhibitor, so that the growth of bacteria can be effectively inhibited, and the probability that part of fungi to be separated is inhibited due to the cooperation of multiple bacteriostatic agents is reduced. The culture medium solves the problems that the existing culture medium for separating the fungi has single components of a carbon source and a nitrogen source, the ratio of the source to the nitrogen source is easy to unbalance, and simultaneously, a plurality of bacteriostats are combined for bacteriostasis, so that the bacteriostasis effect is superposed on part of the fungi, and the effect of separating part of the fungi is poor.
Description
Technical Field
The invention relates to the technical field of medical chemiluminescence immunoassay detection, in particular to a culture medium for separating fungi and yeast-like fungi and a preparation method thereof.
Background
Candida belongs to conditional pathogenic bacteria, exists in skin, oral cavity, vagina, intestinal mucosa and other places, and causes related diseases due to clinical common infection caused by organism hypoimmunity. In order to accurately diagnose the bacterial type of an infected part, a patient part collection sample is usually adopted for culturing clinically, and a clinical laboratory technician selects suspicious bacterial strain types according to clinical symptoms for identification, so that the infected bacterial type of the patient part is judged, and a basis is provided for clinical treatment. Diagnosis of the type of bacteria infected at the site of disease, culture, is currently a viable approach in clinical diagnosis. Clinically, a solid culture medium, a liquid culture medium or a selective culture medium is usually adopted to separate suspicious strains so as to carry out further clinical identification.
Chinese patent publication No. CN109161487A discloses a special culture medium for separating endophytic trichoderma fungi, and a preparation method and application thereof, wherein a carbon source and a nitrogen source of the special culture medium are mainly based on potato and glucose, so that the proportion of the carbon source and the nitrogen source is unbalanced, and meanwhile, chloramphenicol, streptomycin sulfate, Bengal and quintozene are used as bacteriostatic agents, and the bacteriostatic effect of the special culture medium is superposed on part of fungi, so that the effect of separating part of fungi is poor.
Disclosure of Invention
The invention provides a culture medium for separating fungi and yeast-like fungi and a preparation method thereof, aiming at the problems that the existing culture medium for separating fungi has single components of a carbon source and a nitrogen source, the source and nitrogen source ratio is easy to unbalance, and simultaneously, a plurality of bacteriostats are combined for bacteriostasis, so that the bacteriostasis effect is superposed on part of fungi, and the effect of separating part of fungi is poor.
The invention solves the technical problem and adopts the technical scheme that the culture medium for separating the fungi and the yeast-like fungi comprises tryptose peptone, animal peptone, glucose, chloramphenicol, agar and purified water.
Further, the weight parts of the tryptone and the peptone are 5-15 parts, the glucose is 30-50 parts, the agar is 10-20 parts, the chloramphenicol is 0.3-0.7 part, and the purified water is 900-1100 parts.
Further, the weight parts of the tryptone and the peptone are 7-13 parts, the glucose is 35-45 parts, the agar is 12-18 parts, the chloramphenicol is 0.4-0.6 part, and the purified water is 950-1050 parts.
Further, by weight, the tryptone is 10 parts, the animal peptone is 10 parts, the glucose is 40 parts, the agar is 15 parts, the chloramphenicol is 0.5 part, and the purified water is 1000 parts.
The application also provides a preparation method of the culture medium for separating the fungi and the yeast-like fungi, which comprises the following steps:
s1, preparing a reagent, namely uniformly mixing a proper amount of tryptone, animal peptone, glucose, chloramphenicol and agar, and adding purified water to prepare a mixed reagent;
s2, performing moist heat sterilization, namely placing the mixed reagent in a moist heat environment for sterilization treatment to prepare a sterilized reagent;
s3, filling, namely filling the sterilized reagent into a sterilized culture bottle;
s4, drying and sealing, drying the filled culture bottle, and sealing by a gland;
s5, spraying a code to seal the membrane, spraying an information code on the surface of the culture bottle with the gland and sealing the membrane;
s6, labeling, wherein a label is attached to the surface of the culture bottle after the membrane is sealed.
Further, in S2, heat is applied at 120 to 130 ℃ for 30min to perform moist heat sterilization.
Optionally, after S6, there is a sampling detection, where the sampling detection includes the following steps:
s7, extracting a certain amount of culture medium from each batch in the label-pasted culture bottle, and checking whether the culture medium is qualified or not;
and S8, warehousing the batches meeting the qualification rate, and destroying the batches not meeting the qualification rate.
The beneficial effects of the invention at least comprise one of the following;
1. by using tryptone, animal peptone and glucose to provide proper carbon sources and nitrogen sources for the microorganisms, the proportion of the whole carbon sources and the nitrogen sources is controllable, and the glucose content is higher, so that the pH of the whole culture medium tends to 5.5-6.5, and the growth of fungi and yeast-like fungi is facilitated.
2. Meanwhile, single chloramphenicol is used as a bacteria inhibitor, so that the growth of bacteria can be effectively inhibited, and the probability that part of fungi to be separated is inhibited due to the cooperation of multiple bacteriostatic agents is reduced.
3. The culture medium solves the problems that the existing culture medium for separating the fungi has single components of a carbon source and a nitrogen source, the ratio of the source to the nitrogen source is easy to unbalance, and simultaneously, a plurality of bacteriostats are combined for bacteriostasis, so that the bacteriostasis effect is superposed on part of the fungi, and the effect of separating part of the fungi is poor.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail with reference to the following embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the scope of the invention.
Example 1
A method for preparing a culture medium for separating fungi and yeast-like fungi comprises the following steps:
s1, preparing a reagent, namely uniformly mixing 5 parts of tryptone, 5 parts of animal peptone, 30 parts of glucose, 0.3 part of chloramphenicol and 10 parts of agar in parts by weight, and adding 900 parts of purified water into the mixture to prepare a mixed reagent;
s2, performing moist heat sterilization, namely placing the mixed reagent in a moist heat environment, and heating the mixed reagent at the temperature of 120-130 ℃ for 30min to perform moist heat sterilization to prepare a sterilized reagent;
s3, filling, namely filling the sterilized reagent into a sterilized culture bottle;
s4, drying and sealing, drying the filled culture bottle, and sealing by a gland;
s5, spraying a code to seal the membrane, spraying an information code on the surface of the culture bottle with the gland and sealing the membrane;
s6, labeling, wherein a label is attached to the surface of the culture bottle after the membrane is sealed.
In use, the tryptone, the animal peptone and the glucose are used for providing proper carbon sources and nitrogen sources for microorganisms, the proportion of the whole carbon sources and nitrogen sources is controllable, and the glucose content is more, so that the pH of the whole culture medium tends to 5.5-6.5, and the growth of fungi and yeast-like fungi is facilitated. Meanwhile, single chloramphenicol is used as a bacteria inhibitor, so that the growth of bacteria can be effectively inhibited, and the probability that part of fungi to be separated is inhibited due to the cooperation of multiple bacteriostatic agents is reduced. The culture medium solves the problems that the existing culture medium for separating the fungi has single components of a carbon source and a nitrogen source, the ratio of the source to the nitrogen source is easy to unbalance, and simultaneously, a plurality of bacteriostats are combined for bacteriostasis, so that the bacteriostasis effect is superposed on part of the fungi, and the effect of separating part of the fungi is poor.
Wherein the tryptone is tryptone.
Example 2
A method for preparing a culture medium for separating fungi and yeast-like fungi comprises the following steps:
s1, preparing a reagent, namely, taking 15 parts of tryptone, 15 parts of animal peptone, 50 parts of glucose, 0.7 part of chloramphenicol and 20 parts of agar according to parts by weight, uniformly mixing, and adding 1100 parts of purified water into the mixture to prepare a mixed reagent;
s2, performing moist heat sterilization, namely placing the mixed reagent in a moist heat environment, and heating the mixed reagent at the temperature of 120-130 ℃ for 30min to perform moist heat sterilization to prepare a sterilized reagent;
s3, filling, namely filling the sterilized reagent into a sterilized culture bottle;
s4, drying and sealing, drying the filled culture bottle, and sealing by a gland;
s5, spraying a code to seal the membrane, spraying an information code on the surface of the culture bottle with the gland and sealing the membrane;
s6, labeling, wherein a label is attached to the surface of the culture bottle after the membrane is sealed.
In use, the tryptone, the animal peptone and the glucose are used for providing proper carbon sources and nitrogen sources for microorganisms, the proportion of the whole carbon sources and nitrogen sources is controllable, and the glucose content is more, so that the pH of the whole culture medium tends to 5.5-6.5, and the growth of fungi and yeast-like fungi is facilitated. Meanwhile, single chloramphenicol is used as a bacteria inhibitor, so that the growth of bacteria can be effectively inhibited, and the probability that part of fungi to be separated is inhibited due to the cooperation of multiple bacteriostatic agents is reduced. The culture medium solves the problems that the existing culture medium for separating the fungi has single components of a carbon source and a nitrogen source, the ratio of the source to the nitrogen source is easy to unbalance, and simultaneously, a plurality of bacteriostats are combined for bacteriostasis, so that the bacteriostasis effect is superposed on part of the fungi, and the effect of separating part of the fungi is poor.
Wherein the tryptone is tryptone.
Example 3
A method for preparing a culture medium for separating fungi and yeast-like fungi comprises the following steps:
s1, preparing a reagent, namely, taking 7 parts of tryptone, 7 parts of animal peptone, 35 parts of glucose, 0.4 part of chloramphenicol and 12 parts of agar according to parts by weight, uniformly mixing, and adding 950 parts of purified water into the mixture to prepare a mixed reagent;
s2, performing moist heat sterilization, namely placing the mixed reagent in a moist heat environment, and heating the mixed reagent at the temperature of 120-130 ℃ for 30min to perform moist heat sterilization to prepare a sterilized reagent;
s3, filling, namely filling the sterilized reagent into a sterilized culture bottle;
s4, drying and sealing, drying the filled culture bottle, and sealing by a gland;
s5, spraying a code to seal the membrane, spraying an information code on the surface of the culture bottle with the gland and sealing the membrane;
s6, labeling, wherein a label is attached to the surface of the culture bottle after the membrane is sealed.
In use, the tryptone, the animal peptone and the glucose are used for providing proper carbon sources and nitrogen sources for microorganisms, the proportion of the whole carbon sources and nitrogen sources is controllable, and the glucose content is more, so that the pH of the whole culture medium tends to 5.5-6.5, and the growth of fungi and yeast-like fungi is facilitated. Meanwhile, single chloramphenicol is used as a bacteria inhibitor, so that the growth of bacteria can be effectively inhibited, and the probability that part of fungi to be separated is inhibited due to the cooperation of multiple bacteriostatic agents is reduced. The culture medium solves the problems that the existing culture medium for separating the fungi has single components of a carbon source and a nitrogen source, the ratio of the source to the nitrogen source is easy to unbalance, and simultaneously, a plurality of bacteriostats are combined for bacteriostasis, so that the bacteriostasis effect is superposed on part of the fungi, and the effect of separating part of the fungi is poor.
Wherein the tryptone is tryptone.
Example 4
A method for preparing a culture medium for separating fungi and yeast-like fungi comprises the following steps:
s1, preparing a preparation agent, namely, taking 13 parts of tryptone, 13 parts of animal peptone, 45 parts of glucose, 0.6 part of chloramphenicol and 18 parts of agar according to parts by weight, uniformly mixing, and adding 1050 parts of purified water into the mixture to prepare a mixed reagent;
s2, performing moist heat sterilization, namely placing the mixed reagent in a moist heat environment, and heating the mixed reagent at the temperature of 120-130 ℃ for 30min to perform moist heat sterilization to prepare a sterilized reagent;
s3, filling, namely filling the sterilized reagent into a sterilized culture bottle;
s4, drying and sealing, drying the filled culture bottle, and sealing by a gland;
s5, spraying a code to seal the membrane, spraying an information code on the surface of the culture bottle with the gland and sealing the membrane;
s6, labeling, wherein a label is attached to the surface of the culture bottle after the membrane is sealed.
In use, the tryptone, the animal peptone and the glucose are used for providing proper carbon sources and nitrogen sources for microorganisms, the proportion of the whole carbon sources and nitrogen sources is controllable, and the glucose content is more, so that the pH of the whole culture medium tends to 5.5-6.5, and the growth of fungi and yeast-like fungi is facilitated. Meanwhile, single chloramphenicol is used as a bacteria inhibitor, so that the growth of bacteria can be effectively inhibited, and the probability that part of fungi to be separated is inhibited due to the cooperation of multiple bacteriostatic agents is reduced. The culture medium solves the problems that the existing culture medium for separating the fungi has single components of a carbon source and a nitrogen source, the ratio of the source to the nitrogen source is easy to unbalance, and simultaneously, a plurality of bacteriostats are combined for bacteriostasis, so that the bacteriostasis effect is superposed on part of the fungi, and the effect of separating part of the fungi is poor.
Wherein the tryptone is tryptone.
Example 5
A method for preparing a culture medium for separating fungi and yeast-like fungi comprises the following steps:
s1, preparing a reagent, namely, taking 10 parts of tryptone, 10 parts of animal peptone, 40 parts of glucose, 0.5 part of chloramphenicol and 15 parts of agar according to parts by weight, uniformly mixing, and adding 1000 parts of purified water to prepare a mixed reagent;
s2, performing moist heat sterilization, namely placing the mixed reagent in a moist heat environment, and heating the mixed reagent at the temperature of 120-130 ℃ for 30min to perform moist heat sterilization to prepare a sterilized reagent;
s3, filling, namely filling the sterilized reagent into a sterilized culture bottle;
s4, drying and sealing, drying the filled culture bottle, and sealing by a gland;
s5, spraying a code to seal the membrane, spraying an information code on the surface of the culture bottle with the gland and sealing the membrane;
s6, labeling, namely labeling the surface of the culture bottle after the membrane is sealed;
s7, extracting a certain amount of culture medium from each batch in the label-pasted culture bottle, and checking whether the culture medium is qualified or not;
and S8, warehousing the batches meeting the qualification rate, and destroying the batches not meeting the qualification rate.
In use, the tryptone, the animal peptone and the glucose are used for providing proper carbon sources and nitrogen sources for microorganisms, the proportion of the whole carbon sources and nitrogen sources is controllable, and the glucose content is more, so that the pH of the whole culture medium tends to 5.5-6.5, and the growth of fungi and yeast-like fungi is facilitated. Meanwhile, single chloramphenicol is used as a bacteria inhibitor, so that the growth of bacteria can be effectively inhibited, and the probability that part of fungi to be separated is inhibited due to the cooperation of multiple bacteriostatic agents is reduced. The culture medium solves the problems that the existing culture medium for separating the fungi has single components of a carbon source and a nitrogen source, the ratio of the source to the nitrogen source is easy to unbalance, and simultaneously, a plurality of bacteriostats are combined for bacteriostasis, so that the bacteriostasis effect is superposed on part of the fungi, and the effect of separating part of the fungi is poor.
Wherein the tryptone is tryptone. Sampling detection is carried out in the prepared culture medium for separating the fungi and the yeast-like fungi, and unqualified batches are destroyed, so that the overall stability is improved.
Claims (7)
1. A medium for separating fungi and yeast-like fungi, characterized in that: including tryptose, animal peptone, glucose, chloramphenicol, agar and purified water.
2. A culture medium for separating fungi and yeast-like fungi according to claim 1, wherein: the tryptone and casein peptone comprise, by weight, 5-15 parts of tryptone and peptone, 5-15 parts of glucose, 10-20 parts of agar, 0.3-0.7 part of chloramphenicol, and 900-1100 parts of purified water.
3. A culture medium for separating fungi and yeast-like fungi according to claim 2, wherein: the tryptone and casein peptone comprise, by weight, 7-13 parts of tryptone and peptone 7-13 parts of peptone, 35-45 parts of glucose, 12-18 parts of agar, 0.4-0.6 part of chloramphenicol, and 950-1050 parts of purified water.
4. A medium according to claim 3 for the isolation of fungi and yeast-like fungi, characterized in that: according to the weight portion, the tryptone is 10 portions, the animal peptone is 10 portions, the glucose is 40 portions, the agar is 15 portions, the chloramphenicol is 0.5 portion, and the purified water is 1000 portions.
5. A method for preparing a culture medium for separating fungi and yeast-like fungi, which is characterized by comprising the following steps: the method comprises the following steps:
s1, preparing a reagent, namely uniformly mixing a proper amount of tryptone, animal peptone, glucose, chloramphenicol and agar, and adding purified water to prepare a mixed reagent;
s2, performing moist heat sterilization, namely placing the mixed reagent in a moist heat environment for sterilization treatment to prepare a sterilized reagent;
s3, filling, namely filling the sterilized reagent into a sterilized culture bottle;
s4, drying and sealing, drying the filled culture bottle, and sealing by a gland;
s5, spraying a code to seal the membrane, spraying an information code on the surface of the culture bottle with the gland and sealing the membrane;
s6, labeling, wherein a label is attached to the surface of the culture bottle after the membrane is sealed.
6. The method according to claim 5, wherein the culture medium for separating fungi and yeast-like fungi comprises: and S2, heating for 30min at 120-130 ℃ for carrying out damp-heat sterilization.
7. The method according to claim 6, wherein the culture medium for separating fungi and yeast-like fungi comprises: following S6, there is a sample test, which includes the steps of:
s7, extracting a certain amount of culture medium from each batch in the label-pasted culture bottle, and checking whether the culture medium is qualified or not;
and S8, warehousing the batches meeting the qualification rate, and destroying the batches not meeting the qualification rate.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112831538A (en) * | 2021-01-13 | 2021-05-25 | 海南科颜纳生物科技有限公司 | Quick fungus detection reagent |
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