CN1095500C - Combined mycoplasma test reagent and preparing process thereof - Google Patents
Combined mycoplasma test reagent and preparing process thereof Download PDFInfo
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- CN1095500C CN1095500C CN00113758A CN00113758A CN1095500C CN 1095500 C CN1095500 C CN 1095500C CN 00113758 A CN00113758 A CN 00113758A CN 00113758 A CN00113758 A CN 00113758A CN 1095500 C CN1095500 C CN 1095500C
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Abstract
The present invention relates to a reagent used for detecting combined mycoplasmas and a preparing method thereof. The reagent used for detecting combined mycoplasmas is prepared from mixing basic medium, fresh calf serum, fresh yeast liquid, urea, arginine, phenol red and penicillin according to a certain proportion. The reagent used for detecting combined mycoplasmas is not only beneficial to the growth of ureaplasma urealyticum but is also beneficial to the growth of mycoplasma hominis, and the reagent used for detecting combined mycoplasmas contains urea which is decomposed and used as energy sources by ureaplasma urealyticum and arginine which is decomposed into energy by mycoplasma hominis. The present invention has stable performance, intuitionistic judge standard, strong specificity, high detection rate and no need of special equipment, and results can be reported within 24 hours. The present invention provides guarantee for timely and effective treatment for mycoplasma infections.
Description
The invention belongs to field of medicaments, detection reagent of particularly a kind of urogenital tract disease that causes because of mycoplasma and preparation method thereof.
Mycoplasma is the common disease substance of spreading through sex intercourse property urogenital tract disease.Recall rate accounts for venereal disease crowd's 40～70%, and mycoplasma infection can cause diseases such as urodaeum inflammation, prostatitis, pelvic inflammatory disease, male sterility, female acyesis, chorion inflammation and low birthweight infant.At present, more to mycoplasma laboratory diagnosis Study on Technology both at home and abroad, as (Taylor-Robinson D.Clin Lab Sci, 1989 such as specific antibody detection technique, metabolic inhibition test, Southem Blot, PCR method, culture methods; 9 (30): 501-523. and Jensen JS et al.J ClinMicrobiol, 1991; 29 (1): 46-50).The antibody test technology is prone to cross reaction.Metabolic inhibition test need add the growth that specific antibody suppresses mycoplasma, and clinical application is few.The SouthernBlot experimental technique is had relatively high expectations, and is difficult for generally carrying out.False positive and false negative appear in PCR method easily, by the health ministry Clinical Laboratory of ordering to forbid being applied to.And the culture method specificity selective medium that to be the life habit according to mycoplasma adopt detects the method for mycoplasma, each hospital substratum commonly used mostly is single mycoplasma culture medium at present, as single urea mycoplasma culture medium, the single mycoplasma hominis substratum separated.The domestic existing people of single mycoplasma culture medium has declared patent (application number: 96117201.0 contrivers: Wang Heying).M.urealyticum is domestic to have tame unit application to arrive lot number (Shanghai company of Golden Bridge).But the combined mycoplasma substratum does not see that the someone declares patent, and also not seeing so far both at home and abroad has lot number to come out.
The objective of the invention is to overcome the shortcoming of above-mentioned prior art, the reagent and the preparation method of several mycoplasma species have been proposed in a kind of reagent, to detect simultaneously, reached and reduce cost, prevent omission, save patient's spending and reduce purposes such as testing amount, for the timely and effective treatment of mycoplasmosis provides assurance.
Combined mycoplasma test reagent is a kind of liquid nutrient medium, and its preparation method is as follows:
1, substratum: the extraction of fresh beef leach liquor and pig stomach or lung digestive fluid:
(1), the extraction of beef leach liquor: after 1～2 jin of rubbing of fresh beef, added 500～1000ml distilled water immersion 10～25 hours, boiled its filtrate of leaching.
(2), the extraction of pig stomach or lung digestive fluid: get 1～2 jin in fresh pig stomach or lung, rub, add 500～1000ml distilled water, 0.3～2 gram proteolytic enzyme, digested 10～25 hours, boil filtration, get its filtrate.
2, the preparation of basic medium:
In following ratio following material is mixed:
Beef leach liquor 300～600ml
Pig stomach or lung digestive fluid 100～400ml
Sodium-chlor 3～5 grams
Potassium primary phosphate 1～3 gram
Peptone 5～20 grams
After treating that solid chemical material dissolves fully, it is standby to boil after-filtration.
3, the preparation of perfect medium:
In following ratio following material is mixed:
Basic medium 500～900ml
Fresh calf serum 100～500ml
Fresh yeast liquid 50～150ml
Urea element 0.5～2 gram
Arginine 3～15 grams
Phenol red 50～150 milligrams
Penicillin 50～1,500,000 units
After the perfect medium preparation is finished, be combined mycoplasma test reagent.This reagent had both helped the M.urealyticum growth, helped the mycoplasma hominis growth again.And contain promising M.urealyticum and decompose the urea that is utilized as the energy and, discharge NH after urea and arginine are decomposed for mycoplasma hominis is decomposed into the arginine of energy
4 +, reagent pH value is raise, finally make the phenol red indicator in the substratum become redness from yellow.
The present invention can detect mycoplasma hominis and M.urealyticum efficiently simultaneously, compares with single mycoplasma test reagent, is characterized in: stable performance, and judging criterion is directly perceived, high specificity, the recall rate height does not need specific installation, can report the result in 24 hours.This reagent is on probation in 6 tame general hospital, has observed 5000 many cases cases, and effect is fine.For the timely and effective treatment of mycoplasma infection disease provides assurance.
Claims (6)
1. combined mycoplasma test reagent is characterized in that, is mixed in proportion by following substances to form:
Basic medium 500～900ml
Fresh calf serum 100～500ml
Fresh yeast liquid 50～150ml
Urea element 0.5～2 gram
Arginine 3～15 grams
Phenol red 50～150 milligrams
Penicillin 50～1,500,000 units
2. combined mycoplasma test reagent according to claim 1 is characterized in that, said basic medium is mixed in proportion by following substances and forms:
Beef leach liquor 300～600ml
Pig stomach or lung digestive fluid 100～400ml
Sodium-chlor 3～5 grams
Potassium primary phosphate 1～3 gram
Peptone 5～20 grams
3. the preparation method of combined mycoplasma test reagent, it is characterized in that, 500～900ml basic medium, 100～500ml fresh calf serum, 50～150ml fresh yeast liquid, 0.5～2 gram urea, 50～150 milligrams of phenol red being mixed in proportion with 50～1,500,000 unit penicillin of 3～15 gram arginine are formed.
4. the preparation method of combined mycoplasma test reagent according to claim 3, it is characterized in that, said basic medium by: 300～600ml beef leach liquor, 100～400ml pig stomach or lung digestive fluid, 3～5 gram sodium-chlor, 1～3 gram potassium primary phosphate and 5～20 restrain peptones and are mixed in proportion and form.
5, the preparation method of combined mycoplasma test reagent according to claim 4, it is characterized in that, the preparation method of said beef leach liquor is: after fresh beef is rubbed, than 500～1000ml distilled water immersion 10～25 hours, boiled its filtrate of leaching by 1～2 jin of beef.
6, the preparation method of combined mycoplasma test reagent according to claim 4, it is characterized in that, the preparation method of said pig stomach or lung digestive fluid is: after fresh pig stomach or lung rubbing, the ratio that adds 500～1000ml distilled water and 0.3～2 gram proteolytic enzyme in 1～2 jin of pig stomach or lung, digested 10～25 hours, boil filtration, get its filtrate.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN00113758A CN1095500C (en) | 2000-03-15 | 2000-03-15 | Combined mycoplasma test reagent and preparing process thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN00113758A CN1095500C (en) | 2000-03-15 | 2000-03-15 | Combined mycoplasma test reagent and preparing process thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1270227A CN1270227A (en) | 2000-10-18 |
| CN1095500C true CN1095500C (en) | 2002-12-04 |
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN00113758A Expired - Fee Related CN1095500C (en) | 2000-03-15 | 2000-03-15 | Combined mycoplasma test reagent and preparing process thereof |
Country Status (1)
| Country | Link |
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| CN (1) | CN1095500C (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103436468B (en) * | 2013-08-11 | 2015-10-14 | 杭州致远医学检验所有限公司 | Quick growth additive of a kind of helicobacter pylori and preparation method thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5091307A (en) * | 1987-09-29 | 1992-02-25 | Diffusion Bacteriologie Du Var-D.B.V. | Procedure for the counting, detection and identification of mycoplasms in general and urinogenital mycoplasms in particular and a biological medium specially adapted to this effect |
| CN1184155A (en) * | 1996-12-05 | 1998-06-10 | 中国医学科学院皮肤病研究所 | High-efficiency mycoplasma culture medium and its preparation |
-
2000
- 2000-03-15 CN CN00113758A patent/CN1095500C/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5091307A (en) * | 1987-09-29 | 1992-02-25 | Diffusion Bacteriologie Du Var-D.B.V. | Procedure for the counting, detection and identification of mycoplasms in general and urinogenital mycoplasms in particular and a biological medium specially adapted to this effect |
| CN1184155A (en) * | 1996-12-05 | 1998-06-10 | 中国医学科学院皮肤病研究所 | High-efficiency mycoplasma culture medium and its preparation |
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| Publication number | Publication date |
|---|---|
| CN1270227A (en) | 2000-10-18 |
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