CN1098352C - High-efficiency mycoplasma culture medium and its preparation - Google Patents
High-efficiency mycoplasma culture medium and its preparation Download PDFInfo
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- CN1098352C CN1098352C CN96117201A CN96117201A CN1098352C CN 1098352 C CN1098352 C CN 1098352C CN 96117201 A CN96117201 A CN 96117201A CN 96117201 A CN96117201 A CN 96117201A CN 1098352 C CN1098352 C CN 1098352C
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- culture medium
- medium
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- mycoplasma
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Abstract
The present invention provides a high-efficiency mycoplasma culture medium and a preparation method thereof. The high-efficiency mycoplasma culture medium comprises a basal culture medium composed of peptone, the extracting pure powder of fresh beef, sodium chloride, monopotassium phosphate and distilled water. New-born calf serum, fresh yeast extract, 10% carbamide solution, 10% arginine solution and 0.4% phenolsulfonphthalein solution and penicillin are added before the high-efficiency mycoplasma culture medium is used. Agar is also added in a solid culture medium. The high-efficiency mycoplasma culture medium has the characteristics of fast growth and high detection rate under the same experiment conditions.
Description
The present invention relates to a kind of high-efficiency mycoplasma culture medium and preparation method thereof, particularly a kind ofly can make that Ureaplasma urealyticum is grown fast, isolating hypersensitivity and substratum of high specific and preparation method thereof.
Nongonococcal urethritis is the maximum venereal disease of number of the infected in the world, is only second to gonorrhoea.Ureaplasma urealyticum and mycoplasma hominis are one of pathogenic agent that causes nongonococcal urethritis.In the prevention and treatment work of venereal disease, be crucial as detection of mycoplasma to the patient.And in the various laboratory diagnostic methods to mycoplasma, separation and Culture still is used as the most reliable " gold standard ".Mycoplasma culture medium of the prior art has multiple, for example:
U98 urease color test substratum
Basis meat soup
Trysinization meat soup (pulvis) 0.75g
Sodium-chlor 0.50g
Potassium primary phosphate 0.02g
Deionized water 100ml
With 1N HCL adjust pH to 5.5.Through 121 ℃ of autoclavings 15 minutes, separate standby.
Perfect medium (pH6.0)
Basic meat soup (pH5.5) 95ml of sterilization
The not heating horse serum 5ml of sterilization
10% urea stock solution 0.5ml of sterilization
2%L-halfcystine (hydrochloride) 0.1ml of sterilization
1% phenol red solution 0.1ml of sterilization
Potassium penicillin G 100,000 u/ml 1.0ml
For another example:
Fresh beef immersion liquid 100ml
NaCl 0.5g
K
2HPO
4 0.15g
Peptone 1.0g
Urea 0.05g
Phenol red 0.02g
Face with before adding:
Penicillin 1000u/ml
Aseptic foetal calf serum 10ml
Adjust pH to 6.0 ± 0.1
Solid medium: add 1.4% agar
Also have multiple different ingredients such as " A78 differentiate agar " in addition, all can be or be used for the separation and Culture of mycoplasma clinically in scientific research.But the common drawback of prior art is, the speed of growth of mycoplasma in substratum is slower, simultaneously its to detect positive rate also not ideal enough.
At the shortcoming of prior art, task of the present invention provides a kind of new culture medium prescription, and it should have mycoplasma fast growth, isolating susceptibility and the high characteristics of specificity of making.Simultaneously, the present invention also provides the preparation method of this substratum.
The prescription of high-efficiency mycoplasma culture medium is as follows:
At first make basic medium with following ingredients:
1, peptone 9.0-11.0g
2, fresh beef is extracted pure powder 9.0-11.0g
3, sodium-chlor 4.0-6.0g
4, potassium primary phosphate 0.4-0.6g
5, distilled water 1000ml
Add following ingredients before using, make liquid nutrient medium:
6, new-born calf serum 90-110ml
7, fresh yeast immersion liquid 90-110ml
8,10% urea soln or 10% arginine solution 5-15ml
9,0.4% phenol red solution 4-6ml
10, penicillin 500-2000u/ml
In solid medium, also have
11, agar 1.2-1.4%.
The preparation method of above-mentioned substratum is:
One, preparation basic medium
The composition of getting above-mentioned 1-4 mixes, stirs, and makes it to dissolve in fully in the distilled water, filter,
Be basic medium behind the autoclaving, standby.
Two, preparation liquid nutrient medium
The composition that adds above-mentioned 6-10 before using,
Transferring pH with HCL is 6.0, and packing is standby.
Three, preparation solid medium:
Get above-mentioned basic medium, add
Agar 1.2-1.4%
Behind the autoclaving, be cooled to about 50 ℃,
Add the composition of 6-10 one by one with aseptic formality,
Shake up, pour in the sterilization plate, standby.
Substratum provided by the present invention shows with the comparative result of several substratum of the prior art, and under same experiment condition, this high-efficiency mycoplasma culture medium has fast growth, characteristics that recall rate is high.Comparative result sees Table 1, table 2:
Table 1 is separated urine mycoplasma culture medium comparison substratum and is checked the routine number number positive speed of growth
(%)≤24hr (%)＞24hr (%) 〉=48hr (%) the present invention 91 29 (31.9) 25 (86.2) 4 (13.8) 0 (0) existing technology A 91 18 (19.8) 4 (22.2) 8 (44.4) 6 (33.4) the present invention 54 18 (33.3) 17 (94.4) 1 (5.6) 0 (0) existing technology B 54 3 (5.6) 0 (0) 0 (0) 3 (94.4) the present invention 144 53 (36.8) 42 (79.2) 11 (20.8) 0 (0) existing technology C 144 38 (26.4) 14 (36.9) 17 (44.7) 7 (18.4)
Two kinds of mycoplasma hominis substratum of table 2 comparison substratum is checked the routine number number positive speed of growth
(%)≤24hr, (%)＞24hr, (%) 〉=48hr, (%) the present invention 52 4, (7.7) 0, (0) 4, (7.7) 0, (0) existing technology B 52 0, (0) 0, (0) 0, (0) 0, (0) the present invention 144 21, (14.6) 1, (4.8) 12, (57.1) 8, (38.1) existing technology C 144 19, (13.2) 0, (0) 8, (42.1) 11, (57.9)
Now be described further in conjunction with the embodiments.Embodiment 1: the prescription and the preparation method of high-efficiency mycoplasma culture medium are as follows: one, at first make basic medium with following ingredients:
1, peptone 10.0g
2, fresh beef is extracted pure powder 10.0g
3, sodium-chlor 5.0g
4, potassium primary phosphate 0.5g
5, distilled water 1000ml
Get above-mentioned composition and mix, stir, make it to dissolve in fully in the distilled water,
Use filter paper filtering,
Put in the autoclave, standby after 15 pounds of sterilizations in 20 minutes.Two, preparation liquid nutrient medium:
Before the use, get basic medium and add following sterile preparation:
6, new-born calf serum 100ml
7, fresh yeast immersion liquid 100ml
8,10% urea soln (detection Ureaplasma urealyticum) 5-15ml
Or 10% arginine solution (detection mycoplasma hominis) 5-15ml
9,0.4% phenol red solution 5ml
10, penicillin 500-2000u/ml is 6.0 with HCL accent pH, and the packing of sterilization test tube is standby.Three, preparation solid medium:
In the liquid base substratum, add
11, agar 1.2-1.4%
Behind the autoclaving, be cooled to about 50 ℃, add the composition of 6-10 one by one with aseptic formality,
Adjust pH to 6.0 shakes up, and pours in the sterilization plate, solidifies in the rearmounted refrigerator standby.Embodiment 2: compound method and last example are basic identical, but formula rate is as follows: one, at first make basic medium with following ingredients:
1, peptone 9.0g
2, fresh beef is extracted pure powder 9.0g
3, sodium-chlor 4.0g
4, potassium primary phosphate 0.4g
5, distilled water 1000ml
Get the 1-4 composition and mix, stir, make it to dissolve in fully in the distilled water,
Use filter paper filtering,
Put in the autoclaving, standby after 15 pounds of sterilizations in 20 minutes.Two, liquid nutrient medium:
Before the use, get basic medium and add following sterile preparation:
6, new-born calf serum 90ml
7, fresh yeast immersion liquid 90ml
8,10% urea soln (detection Ureaplasma urealyticum) 5-15ml
Or 10% arginine solution 5-15ml
9,0.4% phenol red solution 4ml
10, penicillin 506-2000u/ml is 6.0 with HCL accent pH, and the packing of sterilization test tube is standby.Three, preparation solid medium:
In the liquid base substratum, add
11, agar 1.2-1.4%
Behind the autoclaving, be cooled to about 50 ℃, add the composition of 6-10 one by one with aseptic formality,
Adjust pH to 6.0 shakes up, and pours in the sterilization plate, solidifies in the rearmounted refrigerator standby.Embodiment 3, basic identical with last two examples, but formula rate is as follows: one, at first make basic medium with following ingredients:
1, peptone 11.0g
2, fresh beef is extracted pure powder 11.0g
3, sodium-chlor 6.0g
4, potassium primary phosphate 0.6g
5, distilled water 1000ml
Get above-mentioned 1-4 composition and mix, stir, make it to dissolve in fully in the distilled water,
Use filter paper filtering,
Put in the autoclaving, standby after going out all in 20 minutes through 15 pounds.Two, preparation liquid nutrient medium:
Before the use, get basic medium and add following sterile preparation:
6, new-born calf serum 110ml
7, fresh yeast immersion liquid 110ml
8,10% urea soln (detection Ureaplasma urealyticum) 5-15ml
Or 10% arginine solution (detection mycoplasma hominis) 5-15ml
9,0.4% phenol red solution 6ml
10, penicillin 500-2000u/ml is 6.0 with HCL accent pH, and the packing of sterilization test tube is standby.Three, preparation solid medium:
In the liquid base substratum, add
11, agar 1.2-1.4%
Behind the autoclaving, be cooled to about 50 ℃, add the composition of 6-10 one by one with aseptic formality,
Adjust pH to 6.0 shakes up, and pours in the sterilization plate, solidifies in the rearmounted refrigerator standby.
Claims (3)
1, a kind of high-efficiency mycoplasma culture medium, it is as follows to it is characterized in that filling a prescription: basic medium:
1, peptone 9.0-11.0g
2, fresh beef is extracted pure powder 9.0-11.0g
3, sodium-chlor 4.0-6.0g
4, potassium primary phosphate 0.4-0.6g
5, distilled water 1000ml
Add following ingredients before using and make liquid nutrient medium:
6, new-born calf serum 90-110ml
7, fresh yeast immersion liquid 90-110ml
8,10% urea soln or 10% arginine solution 5-15ml
9,0.4% phenol red solution 4-6ml
10, penicillin 500-2000u/ml
Can add following ingredients again and make solid medium:
11, agar 1.2-1.4%.
2, according to the described high-efficiency mycoplasma culture medium of claim 1, it is characterized in that: prescription limits as follows:
1, peptone 10.0g
2, fresh beef is extracted pure powder 10.0g
3, sodium-chlor 5.0g
4, potassium primary phosphate 0.5g
5, distilled water 1000ml
6, new-born calf serum 100ml
7, fresh yeast immersion liquid 100ml
8,10% urea soln (detection Ureaplasma urealyticum) 5-15ml
Or 10% arginine solution (detection mycoplasma hominis) 5-15ml
9,0.4% phenol red solution 5ml
10, penicillin 500-2000u/ml also has in solid medium
11, agar 1.2-1.4%.
3, a kind of preparation method of high-efficiency mycoplasma culture medium is characterized in that:
At first make basic medium with following ingredients:
1, peptone 9.0-11.0g
2, fresh beef is extracted pure powder 9.0-11.0g
3, sodium-chlor 4.0-6.0g
4, potassium primary phosphate 0.4-0.5g
5, distilled water 1000ml
Get above-mentioned 1-4 composition and mix, stir, make it to dissolve in fully in the distilled water, filter, the sterilization back is standby.The preparation liquid nutrient medium:
Before the use, get above-mentioned basic medium and add following sterile preparation:
6, new-born calf serum 90-110ml
7, fresh yeast immersion liquid 90-110ml
8,10% urea soln (detection Ureaplasma urealyticum) 5-15ml
Or 10% arginine solution 5-15ml
9,0.4% phenol red solution 4-6ml
10, penicillin 500-2000u/ml
Transferring pH is 6.0, and the packing of sterilization test tube is standby, the preparation solid medium:
In basic medium, add
11, agar 1.2-1.4%
Behind the autoclaving, be cooled to about 50 ℃, add the composition of 6-10 one by one with aseptic formality,
Adjust pH to 6.0, standby after solidifying.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN96117201A CN1098352C (en) | 1996-12-05 | 1996-12-05 | High-efficiency mycoplasma culture medium and its preparation |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN96117201A CN1098352C (en) | 1996-12-05 | 1996-12-05 | High-efficiency mycoplasma culture medium and its preparation |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1184155A CN1184155A (en) | 1998-06-10 |
| CN1098352C true CN1098352C (en) | 2003-01-08 |
Family
ID=5124133
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN96117201A Expired - Fee Related CN1098352C (en) | 1996-12-05 | 1996-12-05 | High-efficiency mycoplasma culture medium and its preparation |
Country Status (1)
| Country | Link |
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| CN (1) | CN1098352C (en) |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1095500C (en) * | 2000-03-15 | 2002-12-04 | 黄威权 | Combined mycoplasma test reagent and preparing process thereof |
| CN103849673B (en) * | 2014-03-27 | 2015-03-25 | 武汉菁华时间科技有限公司 | MP (mycoplasma pneumoniae) culture and identification reagent and preparation method thereof |
| CN103937728B (en) * | 2014-05-07 | 2016-05-11 | 石河子大学 | Mycoplasma bovis expands cultivates special culture media (MB-HLS) and preparation method thereof |
| CN111304279A (en) * | 2020-03-09 | 2020-06-19 | 珠海市银科医学工程股份有限公司 | Mycoplasma identification agar culture medium and preparation method thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3883395A (en) * | 1972-03-18 | 1975-05-13 | Behringwerke Ag | Medium for the cultivation of mycoplasms |
| US4473647A (en) * | 1981-02-27 | 1984-09-25 | Amf Inc. | Tissue culture medium |
-
1996
- 1996-12-05 CN CN96117201A patent/CN1098352C/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3883395A (en) * | 1972-03-18 | 1975-05-13 | Behringwerke Ag | Medium for the cultivation of mycoplasms |
| US4473647A (en) * | 1981-02-27 | 1984-09-25 | Amf Inc. | Tissue culture medium |
Also Published As
| Publication number | Publication date |
|---|---|
| CN1184155A (en) | 1998-06-10 |
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Owner name: WANG HEYING; YE SHUNZHANG; SHI MEIQIN Free format text: FORMER NAME OR ADDRESS: WANG HEYING |
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Inventor after: Wang Heying Inventor after: Ye Shunzhang Inventor after: Shi Meiqin Inventor before: Wang Heying |
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