CN105112494B - A kind of combined mycoplasma test reagent and preparation method thereof - Google Patents
A kind of combined mycoplasma test reagent and preparation method thereof Download PDFInfo
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- CN105112494B CN105112494B CN201510512645.8A CN201510512645A CN105112494B CN 105112494 B CN105112494 B CN 105112494B CN 201510512645 A CN201510512645 A CN 201510512645A CN 105112494 B CN105112494 B CN 105112494B
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Abstract
The invention discloses a kind of combined mycoplasma test reagents, are made of following compositions：6.5~10g of Yeast protein peptone, 0.015~0.025g of ampicillin sodium salt, 0.5~1.0g of L arginine, 0.001~0.003g of thallous acetate, 4~12g of agar powder, 0.02~0.1g of urea, 2.25~2.75g of lactoalbumin hydrolysate, 0.01~0.05g of sericin, 0.017~0.023g of chlorophenol red solution, 0.003~0.006g of genipic acid, 0.001~0.01g of toad general glycoside, 10~15ml of bacteriostatic agent.Detection reagent of the present invention can quickly detect solution urea mycoplasma and mycoplasma hominis simultaneously, and detection method is simple, the time is short, selectivity is strong, high sensitivity, recall rate are high.
Description
Technical field
The present invention relates to pharmaceutical technology field field, more particularly to a kind of combined mycoplasma test reagent and its preparation side
Method.
Background technology
Mycoplasma is the common causative of spreading through sex intercourse property genito-urinary disorders.Recall rate account for STD population group 40~
70%, mycoplasma infection can cause Urogenital, prostatitis, pelvic infecton, male sterility, female acyesis, villus adenositis
With the diseases such as low birth weight infant.Mainly decomposing urea mycoplasma and the mycoplasma hominis two related with urogenital infections
Kind, there are about the patients of the non-gonococcal urethritis of 20-30%, are caused by Ureaplasma urealyticum (UU) and mycoplasma hominis (MH)
, be non-gonococcal urethritis and cervicitis second substantially germ.Therefore, the detection of Ureaplasma urealyticum and mycoplasma hominis exists
Clinically have great importance.
At present, detect the method for Ureaplasma urealyticum and mycoplasma hominis have detection of specific antibody method, PCR methods,
Southern Blot methods and isolated culture etc..These methods all respectively have its advantage and disadvantage, wherein, immunological detection method is easy
There is cross reaction, PCR methods are susceptible to false negative and false positive, and Southern Blot methods are more demanding to experimental technique,
Be not suitable for using in hospital.
Clinically common detection method is isolated culture.The domestic reality in relation to Urogenital Mycoplasma culture should
Liquid culture method is mainly used with the identification with studies have shown that mycoplasma.Although liquid culture method is easy to use, quick, easy
Judge, but this method can only infer that mycoplasma whether there is from the change of liquid color, false positive rate is high, and accuracy is not
It is high.And solid culture rule can observe the bacterium colony of mycoplasma hominis and ureaplasma urealyticum by microscope, so as to confirm branch
The presence of substance preferably solves the problems, such as that clinical samples false positive rate is high.But the time of plate method sentence read result compared with
It is long, thus need the time it is longer the urinary tract reproductive tract infection caused by UU and MH could be made a definite diagnosis, be unfavorable for examining for disease
Disconnected and treatment.Therefore, it is necessary to work out a kind of detection that can quickly detect solution urea mycoplasma and mycoplasma hominis simultaneously
Reagent.
The content of the invention
The technical problem to be solved by the present invention is to provide a kind of combined mycoplasma test reagent and preparation method thereof, Neng Goutong
When quickly detect solution urea mycoplasma and mycoplasma hominis, method is simple, the time is short, accuracy is strong, and stability is good.
The technical scheme is that a kind of combined mycoplasma test reagent, is made of following compositions：
Further, the bacteriostatic agent is 18% peppermint water decoction, and peppermint water decoction can effectively inhibit golden yellow Portugal
Grape coccus, the growth of Escherichia coli, Pseudomonas aeruginosa, Candida albicans, the fungi of candida albicans and other miscellaneous bacterias.
Further, the concentration of the chlorophenol red solution is 0.04%, and chlorophenol red solution is indicator, works as training
It supports chlorophenol red in base and becomes red i.e. provable with the presence of M.urealyticum and mycoplasma hominis from yellow.
Wherein, the Yeast protein peptone provides carbon nitrogen source, energy, the vitamin B complex needed for mycoplasma growth；Hydrolysis breast
Albumen and sericin contain various polypeptides, growth factor, hormone etc., these substances are to promoting the breeding of mycoplasma and growing all
It is critically important factor, sericin avoids the pollution that serum is brought, significantly reduces cost, improves cell culture body
The stability of system；Ampicillin sodium salt inhibits harmful virus, is conducive to inhibit other bacterial growths, maintains balanced infiltration
Pressure：Agar powder is the coagulator of culture medium；L-arginine is the energy utilized as mycoplasma hominis decomposition, and urea is as solution
The energy that the decomposition of urea type mycoplasma utilizes；Chlorophenol red solution is indicator, when chlorophenol red becomes from yellow in culture medium
It is i.e. provable with the presence of M.urealyticum and mycoplasma hominis into red；Genipic acid can inhibit gram positive and negative extensively
The growth of bacterium；Toad general glycoside has proteus, Pseudomonas aeruginosa, tetrads, staphylococcus albus, micrococcus catarrhalis inhibition to make
With peppermint water decoction can effectively inhibit staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Candida albicans, white false silk
The fungi of yeast and the growth of other miscellaneous bacterias.
A kind of preparation method of combined mycoplasma test reagent, comprises the following steps：
1) agar powder is dissolved in 200ml distilled water, high pressure sterilization, obtains solution I, keep solution I temperature 45~
It is 50 DEG C, spare；
2) by sericin, Yeast protein peptone, ampicillin sodium salt, L-arginine, thallous acetate, urea, hydrolysis breast
Albumen is added in 500ml distilled water, mixing, is heated up to 45~50 DEG C, after to be dissolved, and impurity screening obtains solution II, standby
With；
3) solution II pH to 6.0~6.3 is adjusted, with pressure steam sterilizer in 112 DEG C of pressuresteam sterilizations, sterilizing duration
35~45min keeps 45~50 DEG C of temperature, spare；
4) solution I with solution II is merged, adds in chlorophenol red solution, genipic acid, toad general glycoside, bacteriostatic agent, add
Enter distilled water to 1500ml, mixing is filtered under aseptic condition by the supermicro filtration membrane that aperture is 0.25 μm, is poured into sterilized petri dishes
In, treat its solidification, 4 DEG C be sealed it is spare.
Compared with prior art, the present invention has the following advantages：(1) culture medium of detection reagent of the present invention it is full of nutrition,
It prepares simple；(2) compared with single mycoplasma test reagent, solution urea mycoplasma can be quickly detected simultaneously and human-like branch is former
Body, selectivity is strong, discoloration is sensitive, recall rate is high, pollution is small, clinical application effect satisfaction；(3) detection method is simple, the time is short,
Accuracy is strong, and high sensitivity, high specificity, stability are good.
Specific embodiment
It is further described the present invention With reference to embodiment.
Embodiment 1：
A kind of combined mycoplasma test reagent, is made of following compositions：
Wherein, the bacteriostatic agent is 18% peppermint water decoction, and peppermint water decoction can effectively inhibit Staphylococcus aureus
Bacterium, the growth of Escherichia coli, Pseudomonas aeruginosa, Candida albicans, the fungi of candida albicans and other miscellaneous bacterias；The dichloro
The concentration of phenolsulfonphthalein solution is 0.04%, and chlorophenol red solution is indicator, when chlorophenol red becomes from yellow in culture medium
It is i.e. provable with the presence of M.urealyticum and mycoplasma hominis into red.
A kind of preparation method of combined mycoplasma test reagent, comprises the following steps：
1) agar powder is dissolved in 200ml distilled water, high pressure sterilization, obtains solution I, keep solution I temperature at 45 DEG C,
It is spare；
2) by sericin, Yeast protein peptone, ampicillin sodium salt, L-arginine, thallous acetate, urea, hydrolysis breast
Albumen is added in 500ml distilled water, mixing, is heated up to 45 DEG C, after to be dissolved, and impurity screening obtains solution II, spare；
3) solution II pH to 6.0 is adjusted, with pressure steam sterilizer in 112 DEG C of pressuresteam sterilizations, sterilizing duration
35min keeps temperature 45 C, spare；
4) solution I with solution II is merged, adds in chlorophenol red solution, genipic acid, toad general glycoside, bacteriostatic agent, add
Enter distilled water to 1500ml, mixing is filtered under aseptic condition by the supermicro filtration membrane that aperture is 0.25 μm, is poured into sterilized petri dishes
In, treat its solidification, 4 DEG C be sealed it is spare.
Embodiment 2：
A kind of combined mycoplasma test reagent, is made of following compositions：
Wherein, the bacteriostatic agent is 18% peppermint water decoction, and peppermint water decoction can effectively inhibit Staphylococcus aureus
Bacterium, the growth of Escherichia coli, Pseudomonas aeruginosa, Candida albicans, the fungi of candida albicans and other miscellaneous bacterias；The dichloro
The concentration of phenolsulfonphthalein solution is 0.04%, and chlorophenol red solution is indicator, when chlorophenol red becomes from yellow in culture medium
It is i.e. provable with the presence of M.urealyticum and mycoplasma hominis into red.
A kind of preparation method of combined mycoplasma test reagent, comprises the following steps：
1) agar powder is dissolved in 200ml distilled water, high pressure sterilization, obtains solution I, keep solution I temperature 47.5
DEG C, it is spare；
2) by sericin, Yeast protein peptone, ampicillin sodium salt, L-arginine, thallous acetate, urea, hydrolysis breast
Albumen is added in 500ml distilled water, mixing, is heated up to 47.5 DEG C, after to be dissolved, and impurity screening obtains solution II, spare；
3) solution II pH to 6.1 is adjusted, with pressure steam sterilizer in 112 DEG C of pressuresteam sterilizations, sterilizing duration
40min keeps 47.5 DEG C of temperature, spare；
4) solution I with solution II is merged, adds in chlorophenol red solution, genipic acid, toad general glycoside, bacteriostatic agent, add
Enter distilled water to 1500ml, mixing is filtered under aseptic condition by the supermicro filtration membrane that aperture is 0.25 μm, is poured into sterilized petri dishes
In, treat its solidification, 4 DEG C be sealed it is spare.
Embodiment 3：
A kind of combined mycoplasma test reagent, is made of following compositions：
Wherein, the bacteriostatic agent is 18% peppermint water decoction, and peppermint water decoction can effectively inhibit Staphylococcus aureus
Bacterium, the growth of Escherichia coli, Pseudomonas aeruginosa, Candida albicans, the fungi of candida albicans and other miscellaneous bacterias；The dichloro
The concentration of phenolsulfonphthalein solution is 0.04%, and chlorophenol red solution is indicator, when chlorophenol red becomes from yellow in culture medium
It is i.e. provable with the presence of M.urealyticum and mycoplasma hominis into red.
A kind of preparation method of combined mycoplasma test reagent, comprises the following steps：
1) agar powder is dissolved in 200ml distilled water, high pressure sterilization, obtains solution I, keep solution I temperature at 50 DEG C,
It is spare；
2) by sericin, Yeast protein peptone, ampicillin sodium salt, L-arginine, thallous acetate, urea, hydrolysis breast
Albumen is added in 500ml distilled water, mixing, is heated up to 50 DEG C, after to be dissolved, and impurity screening obtains solution II, spare；
3) solution II pH to 6.3 is adjusted, with pressure steam sterilizer in 112 DEG C of pressuresteam sterilizations, sterilizing duration
45min keeps temperature 50 C, spare；
4) solution I with solution II is merged, adds in chlorophenol red solution, genipic acid, toad general glycoside, bacteriostatic agent, add
Enter distilled water to 1500ml, mixing is filtered under aseptic condition by the supermicro filtration membrane that aperture is 0.25 μm, is poured into sterilized petri dishes
In, treat its solidification, 4 DEG C be sealed it is spare.
Clinical practice situation：
This reagent is tried out in 20 general hospital, observes 6000 many cases cases, and effect is fine, is mycoplasma infection
The timely and effective treatment of disease provides guarantee.
Afterwards it should be noted that：The above embodiments are merely illustrative of the technical solutions of the present invention, rather than its limitations；Although ginseng
The present invention is described in detail according to previous embodiment, it will be understood by those of ordinary skill in the art that：It still can be with
It modifies to the technical solution recorded in previous embodiment or equivalent substitution is carried out to which part technical characteristic；And this
A little modifications are replaced, and the essence of appropriate technical solution is not made to depart from the spirit and scope of technical solution of the embodiment of the present invention.
Claims (1)
1. a kind of combined mycoplasma test reagent, which is characterized in that be made of following compositions：
The bacteriostatic agent is 18% peppermint water decoction；
The concentration of the chlorophenol red solution is 0.04%；
A kind of preparation method of combined mycoplasma test reagent, comprises the following steps：
1) agar powder is dissolved in 200ml distilled water, high pressure sterilization, obtains solution I, keep solution I temperature at 45~50 DEG C,
It is spare；
2) by sericin, Yeast protein peptone, ampicillin sodium salt, L-arginine, thallous acetate, urea, lactoalbumin hydrolysate
It is added in 500ml distilled water, mixing, heats, after to be dissolved, impurity screening obtains solution II, spare；
3) solution II pH to 6.0~6.3 is adjusted, pressuresteam sterilization keeps 45~50 DEG C of temperature, spare；
4) solution I with solution II is merged, adds in chlorophenol red solution, genipic acid, toad general glycoside, bacteriostatic agent, added in and steam
Distilled water is to 1500ml, mixing, is filtered under aseptic condition through supermicro filtration membrane, is poured into sterilized petri dishes, treats its solidification, low temperature seal
It saves backup.
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| CN201510512645.8A CN105112494B (en) | 2015-08-18 | 2015-08-18 | A kind of combined mycoplasma test reagent and preparation method thereof |
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| CN201510512645.8A CN105112494B (en) | 2015-08-18 | 2015-08-18 | A kind of combined mycoplasma test reagent and preparation method thereof |
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| CN105112494A CN105112494A (en) | 2015-12-02 |
| CN105112494B true CN105112494B (en) | 2018-05-18 |
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2015
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Non-Patent Citations (3)
| Title |
|---|
| 溶脲脲原体培养基的改良研究;丁建中等;《湖北省卫生职工医学院学报》;19991231(第3期);1-2 * |
| 自制解脲与人型支原体培养基的实验研究;陈芳等;《中国微生态学杂志》;20041231;第16卷(第6期);373、375 * |
| 解脲支原体液体培养方法的改进;刘华等;《中国优生与遗传杂志》;20001231;第8卷(第2期);105、114 * |
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