CN103757112A - Mycobacterium separation and culture kit and testing method thereof - Google Patents
Mycobacterium separation and culture kit and testing method thereof Download PDFInfo
- Publication number
- CN103757112A CN103757112A CN201410019032.6A CN201410019032A CN103757112A CN 103757112 A CN103757112 A CN 103757112A CN 201410019032 A CN201410019032 A CN 201410019032A CN 103757112 A CN103757112 A CN 103757112A
- Authority
- CN
- China
- Prior art keywords
- mycobacterium
- separation
- medium
- test kit
- culture
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
- C12Q1/04—Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor
Abstract
The invention discloses and provides a mycobacterium separation and culture kit and a testing method thereof which have the advantages that the cost is low, the positive detectable rate of mycobacteria in a sample to be detected can be increased, the detection time can be shortened, and special devices and equipment are not required. The kit disclosed by the invention comprises a box body (1), wherein the box body (1) contains a solid culture medium (2), a liquid culture medium (3), a sterile diluent (4) and a competitor inhibitor (5). The testing method comprises the steps of pre-treating the sample to be detected; diluting the competitor inhibitor; adding the competitor inhibitor into the liquid culture medium, and then, adding the liquid culture medium into the solid culture medium, so as to form a double-phase culture medium; adding the treated sample to be detected into the culture medium and incubating; observing a culture result, and judging whether the sample contains the mycobacteria or not. The kit and the testing method thereof are applied to the field of medical detection on the separation and culture of the mycobacteria.
Description
Technical field
The present invention relates to a point a kind of test kit, relate in particular to a kind of branch bacillus separation and Culture test kit and testing method thereof.
Background technology
Tuberculosis is the respiratory infectious disease of serious harm HUMAN HEALTH, and China is one of the high burden of 22, whole world tuberculosis country.According to the 5th the tuberculosis epidemiology investigation result in the whole nation in 2010, estimation China total man group active tuberculosis morbidity is 4,59/,100,000, and wherein infectivity pulmonary tuberculosis morbidity rate is 66/,100,000.Improve recall rate lungy and shorten detection time, early stage discovery tubercular more in time, is treated tubercular timely and effectively, for preventing and controlling propagation lungy, has great importance.The separation and Culture of tubercule bacillus is one of important evidence of diagnosis of tuberculosis, culture method remains objective, the most important and the most most scientific detection method of tuberculosis at present, " gold standard " of infected by microbes diagnosis is culture method, the bacteriological detection of tuberculosis patient is diagnosis of tuberculosis, treatment and the reliable means of evaluating result for the treatment of, is applicable to each unit.The Russell medium generally using clinically for a long time, because its nutritive ingredient is fairly simple, particularly grow the slowly separation and Culture speed of mycobacterium of mycobacterium is slow, and on the low side to the separation and Culture positive rate of the mycobacterium tuberculosis in clinical samples.The World Health Organization advocates energetically and adopts liquid nutrient medium to carry out the detection of mycobacterium separation and Culture.But reagent dependence on import, for example BACTEC-960 system or BacT/ALERT 3D system, its advantage is that nutritive ingredient is high, cultivation speed is fast, shortcoming is expensive, and intuitively observations needs specific apparatus (equipment and substratum all need import); And China tubercular is mainly distributed in the poor areas of economic condition such as rural area, side area, mostly cannot bear quick liquid and cultivate the testing cost of instrument detection system costliness, therefore for our tuberculosis epidemic situation feature, be badly in need of the inexpensive mycobacterium tuberculosis culture of isolated reagent that exploitation not only can improve positive rate but also can shorten detection time, but at present domesticly can be used for clinical market-oriented test kit and also do not have.
Summary of the invention
Technical problem to be solved by this invention is to overcome the deficiencies in the prior art, provide a kind of cheap, both can improve the positive rate of the mycobacterium in sample to be checked, can shorten detection time again, and without mycobacterium separation and Culture test kit and the testing method thereof of special instruments and equipment.
The technical scheme that mycobacterium separation and Culture test kit of the present invention adopts is: described test kit comprises box body, includes solid medium, liquid nutrient medium, sterile diluent and miscellaneous bacteria inhibitor on described box body.
Described solid medium is traditional modified Russell medium component.
Described liquid nutrient medium is comprised of basic medium, growth promoter, fungistat and developer.
Described sterile diluent is comprised of sylvite and sodium salt sterile solution.
Described miscellaneous bacteria inhibitor is comprised of the freeze-dried preparation of PXB, amphotericin B, Nalidixic Acid, azlocillin and trimethoprim.
Described developer is phenol red.
Described growth promoter is microbiotic system.
The technical scheme that the testing method of mycobacterium separation and Culture test kit of the present invention adopts is that the method comprises the following steps:
(1) sample to be checked is carried out to antipollution liquefaction processing;
(2) take out freeze-drying miscellaneous bacteria inhibitor, the bottle cap of outwarding winding, all adds sterile diluent and fully shakes up afterwards;
(3) take out solid medium and liquid nutrient medium, make it before inoculation sample to be checked, approach room temperature, draw the miscellaneous bacteria inhibitor mixing and add liquid nutrient medium, fully mix;
(4) get the liquid nutrient medium mixing, add solid medium, obtain biphasic culture;
(5) above-mentioned steps (1) the sample 0.1ml to be checked after treatment that learnt from else's experience is inoculated in the substratum of processing through step (4) under aseptic condition, puts 37 ℃ of incubations;
(6) inoculate latter 3 days, 7 days each observationss once, later twice of observations weekly; Being cultured to 6 weeks solid culture primary surfaces is not still any change as cultivating feminine gender without any colony growth or liquid nutrient medium; In culture medium slant, occur bacterium colony, or it is positive in liquid medium within, to occur that red or pink particulate state is grown to cultivation.
Further, the blending ratio of basic medium, growth promoter, fungistat and the developer in described liquid nutrient medium is 900 ︰ 100 ︰ 1 ︰ 1.
The invention has the beneficial effects as follows: test kit of the present invention provides complete matched reagent and teachings, clinical user is carried out the detection of mycobacterium in sample according to specification sheets; This test kit has had the two-fold advantage of liquid nutrient medium and Russell medium concurrently, liquid advantage and solid advantage embody a pipe, both had and shortened detection time, the advantage that positive rate is high, there is again the advantage that can observe mycobacterium settlement pattern, and do not need special plant and instrument, be adapted at the area that China medical institutions at different levels, particularly basic unit and resource are relatively deficient and promote the use of.
Accompanying drawing explanation
The easy structure schematic diagram of test kit described in Fig. 1.
Embodiment
Test kit of the present invention comprises box body 1, includes solid medium 2, liquid nutrient medium 3, sterile diluent 4 and miscellaneous bacteria inhibitor 5 on described box body 1, specific as follows.
1, solid medium is produced according to traditional modified Russell medium, filling for 8ml/ props up with the smooth clear polycarbonate cylindrical tube of 80 × 25mm, by steam constant temperature case 50min, 85 ℃, solidifies.Be packaged as 24/box.
2, liquid nutrient medium mixes basic medium, growth promoter, fungistat and developer according to the ratio of 900:100:1:1 after, filling for 2.5ml/ props up with the brown glass screw socket bottle of 5ml, totally 24/box.Described developer is phenol red, and described growth promoter is conventional microbiotic system.
3, sterile diluent is sylvite, the sodium salt sterile solution that 3ml/ props up, filling with antitheft drop bottle, 1/box.
4, miscellaneous bacteria inhibitor is the freeze-dried preparation of PXB, amphotericin B, Nalidixic Acid, azlocillin and trimethoprim, the admixing medical solutions of PXB, amphotericin B, Nalidixic Acid, azlocillin, trimethoprim is made to 1/box by vacuum freeze-drying technique.
5, the detection that adopts mycobacterium separation and Culture test kit to carry out mycobacterium is carried out according to the following steps:
(1) pre-treatment of sample to be checked is with reference to carrying out antipollution liquefaction processing with centrifugal in the alkaline purification in " diagnosis of tuberculosis laboratory inspection rules ";
(2) take out freeze-drying miscellaneous bacteria inhibitor, the bottle cap of outwarding winding, all adds 1 bottle of sterile diluent and fully shakes up afterwards.
(3) take out solid medium and liquid nutrient medium, make it before inoculation sample to be checked, approach room temperature, the miscellaneous bacteria inhibitor that absorption 100ul mixes adds the liquid nutrient medium of 1 bottle of 2.5ml, fully mixes.
(4) get the liquid nutrient medium mixing, add 8ml solid medium, be biphasic culture.
(5) step (1) the sample 0.1ml to be checked after treatment that learns from else's experience is inoculated in step (4) substratum after treatment under aseptic condition, puts 37 ℃ of incubations.
(6) inoculate latter 3 days, 7 days each observationss once, later twice of observations weekly.While being cultured to 10 days, in liquid medium within, there is red or the pink particulate state grown cultures positive.
Concrete steps are: sterile diluent is all joined in miscellaneous bacteria inhibitor and fully shaken up; The miscellaneous bacteria inhibitor that absorption 100ul mixes adds liquid nutrient medium, fully mixes; Get the liquid nutrient medium mixing, add solid medium, be biphasic culture; Inoculate sample to be checked to biphasic culture, inclination substratum makes liquid cover solid culture primary surface completely, puts 37 ℃ of incubation 48h; After hatching 48h, upright substratum and continuation are cultivated, observe cultivation results.
Mycobacterium separation and Culture test kit of the present invention is based on a kind of novel culture medium that can promote mycobacterium Fast Growth, and this substratum is comprised of solid and liquid two parts.Solid part contains general modified Russell medium all the components both at home and abroad, liquid portion is comprised of basic medium, growth promoter, fungistat and developer, this substratum can optionally promote the Fast Growth of mycobacterium, can suppress again other varied bacteria growing; Mycobacterium can form typical bacterium colony on the solid inclined-plane of substratum, simultaneously again because contain developer, presents pink or the growth of red granules shape in liquid medium within, very easily observes, and contributes to the preliminary evaluation of mycobacterium.
The present invention is applied to the field of medical examination of mycobacterium separation and Culture.
Claims (9)
1. a mycobacterium separation and Culture test kit, it is characterized in that: described mycobacterium separation and Culture test kit comprises box body (1), on described box body (1), include solid medium (2), liquid nutrient medium (3), sterile diluent (4) and miscellaneous bacteria inhibitor (5).
2. mycobacterium separation and Culture test kit according to claim 1, is characterized in that: described solid medium (2) is traditional modified Russell medium component.
3. mycobacterium separation and Culture test kit according to claim 1, is characterized in that: described liquid nutrient medium (3) is comprised of basic medium, growth promoter, fungistat and developer.
4. mycobacterium separation and Culture test kit according to claim 1, is characterized in that: described sterile diluent (4) is comprised of sylvite and sodium salt sterile solution.
5. mycobacterium separation and Culture test kit according to claim 1, is characterized in that: described miscellaneous bacteria inhibitor (5) is comprised of the freeze-dried preparation of PXB, amphotericin B, Nalidixic Acid, azlocillin and trimethoprim.
6. mycobacterium separation and Culture test kit according to claim 3, is characterized in that: described developer is phenol red.
7. mycobacterium separation and Culture test kit according to claim 3, is characterized in that: described growth promoter is microbiotic system.
8. a testing method of utilizing mycobacterium separation and Culture test kit as claimed in claim 1 to carry out mycobacterium detection, is characterized in that, the method comprises the following steps:
Sample to be checked is carried out to antipollution liquefaction processing;
Take out freeze-drying miscellaneous bacteria inhibitor, the bottle cap of outwarding winding, all adds sterile diluent and fully shakes up afterwards;
Take out solid medium and liquid nutrient medium, make it before inoculation sample to be checked, approach room temperature, draw the miscellaneous bacteria inhibitor mixing and add liquid nutrient medium, fully mix;
Get the liquid nutrient medium mixing, add solid medium, obtain biphasic culture;
The above-mentioned steps of learning from else's experience (1) sample 0.1ml to be checked after treatment is inoculated in the substratum of processing through step (4) under aseptic condition, puts 37 ℃ of incubations;
Inoculate latter 3 days, 7 days each observationss once, later twice of observations weekly; Being cultured to 6 weeks solid culture primary surfaces is not still any change as cultivating feminine gender without any colony growth or liquid nutrient medium; In culture medium slant, occur bacterium colony, or it is positive in liquid medium within, to occur that red or pink particulate state is grown to cultivation.
9. the testing method of mycobacterium separation and Culture test kit according to claim 8, is characterized in that: the blending ratio of basic medium, growth promoter, fungistat and developer in described liquid nutrient medium is 900:100:1:1.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410019032.6A CN103757112A (en) | 2014-01-15 | 2014-01-15 | Mycobacterium separation and culture kit and testing method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410019032.6A CN103757112A (en) | 2014-01-15 | 2014-01-15 | Mycobacterium separation and culture kit and testing method thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN103757112A true CN103757112A (en) | 2014-04-30 |
Family
ID=50524427
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410019032.6A Pending CN103757112A (en) | 2014-01-15 | 2014-01-15 | Mycobacterium separation and culture kit and testing method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103757112A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104450860A (en) * | 2013-09-17 | 2015-03-25 | 上海市肺科医院 | Pneumonia mycoplasma medium |
| CN106635800A (en) * | 2016-10-31 | 2017-05-10 | 珠海市银科医学工程股份有限公司 | Quick separation and culture method for mycobacteria |
| CN107389666A (en) * | 2017-07-17 | 2017-11-24 | 长沙金域医学检验所有限公司 | A kind of biochemical luminesceence analysis calibrating method |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1699592A (en) * | 2005-04-29 | 2005-11-23 | 胡忠义 | Two phase Roe's culture medium and preparation method thereof |
-
2014
- 2014-01-15 CN CN201410019032.6A patent/CN103757112A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1699592A (en) * | 2005-04-29 | 2005-11-23 | 胡忠义 | Two phase Roe's culture medium and preparation method thereof |
Non-Patent Citations (2)
| Title |
|---|
| 刘华等: "一种快速培养分枝杆菌生长指示(MGIT)系统应用评价", 《实用医院临床杂志》 * |
| 金法祥等: "结核分枝杆菌快速培养", 《临床检验杂志》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104450860A (en) * | 2013-09-17 | 2015-03-25 | 上海市肺科医院 | Pneumonia mycoplasma medium |
| CN104450860B (en) * | 2013-09-17 | 2018-01-02 | 上海市肺科医院 | A kind of mycoplasma pneumoniae culture medium |
| CN106635800A (en) * | 2016-10-31 | 2017-05-10 | 珠海市银科医学工程股份有限公司 | Quick separation and culture method for mycobacteria |
| CN107389666A (en) * | 2017-07-17 | 2017-11-24 | 长沙金域医学检验所有限公司 | A kind of biochemical luminesceence analysis calibrating method |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Bhalla et al. | Methods of phenotypic identification of non-tuberculous mycobacteria | |
| CN100580091C (en) | Zoonosis tuberculosis fluorescence PCR rapid diagnosis kit | |
| CN102177249B (en) | For the reaction culture medium of staphylococcus aureus | |
| CN104611403A (en) | Food and tableware coliform bacteria rapid detection scrip preparation and application | |
| CN104105795A (en) | Method of detecting a salmonella microorganism | |
| CN103757112A (en) | Mycobacterium separation and culture kit and testing method thereof | |
| CN102154472A (en) | Quantitative detection method of salmonella living body in water | |
| Zarzour et al. | Evaluation of three test procedures for identification of Staphylococcus aureus from clinical sources | |
| US10550437B2 (en) | Clostridium difficile culture medium | |
| Lejbkowicz et al. | Recovery of Kingella kingae from blood and synovial fluid of two pediatric patients by using the BacT/Alert system | |
| JP2017108721A (en) | Long-term storable culture medium for culturing obligate anaerobes or microaerophilic bacteria in aerobic environment, and detection method of obligate anaerobes or microaerophilic bacteria using the same culture medium | |
| CN103308360B (en) | The viscosity reduction equipment of toughness clinical memory qu antized table and viscosity reducing process | |
| CN106755274B (en) | Separation and identification method of swine escherichia coli | |
| CN109627330A (en) | A kind of porcine pseudorabies virus high-titer positive serum preparation method | |
| CN104498381B (en) | A kind of Huizhou Autoinducer and application thereof | |
| CN105420341A (en) | B hemolytic streptococcus screening reagent, preparation method thereof and B hemolytic streptococcus screening method | |
| CN105296406A (en) | Culture method and inactivation method of haemophilus parasuis | |
| CN106635800A (en) | Quick separation and culture method for mycobacteria | |
| CN109536568A (en) | A kind of quick detection produces method, kit and its application of carbapenem enzyme bacterial strain | |
| CN106167771B (en) | Bacillus megaterium D122, microbial inoculum thereof and preparation method of microbial inoculum | |
| CN108956247A (en) | A kind of staphylococcus saprophyticus serum antibody slide agglutination detection kit and its application | |
| Upton et al. | Disappointing performance of rapid antigen detection tests for group A streptococcus in the Auckland school-based sore throat programme | |
| WO2011119739A1 (en) | Method and medium for accelerating target analyte growth in a test sample | |
| CN1321193C (en) | Kit for quick identification of active mycobacteria | |
| CN108368534A (en) | Measurement for determining antibiotic in waste |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20140430 |
|
| RJ01 | Rejection of invention patent application after publication |