CN110283882A - There are the methods of mycoplasma pneumoniae in a kind of quick detection clinical sample - Google Patents
There are the methods of mycoplasma pneumoniae in a kind of quick detection clinical sample Download PDFInfo
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Abstract
The present invention discloses in a kind of quickly detection clinical sample that there are the methods of mycoplasma pneumoniae, by detecting the activity for being not present in human body cell but being present in the generation enzyme of the ATP in bovis cells, detect in clinical sample whether contain mycoplasma pneumoniae, to judge whether it is mycoplasma pneumoniae infection.The method of the present invention includes contacting after eluting clinical sample, respectively with reagent I and reagent II, it is measured respectively and the ATP signal in two mixtures again after incubation, and calculate signal ratio or net signal, infer whether contain mycoplasma pneumoniae in clinical sample with this, and then judges whether it is mycoplasma pneumoniae infection.Reagent I contains ATP detection ingredient, and reagent II the reagent I, II is the biochemiluminescence reagent based on luciferase in addition to that can generate containing ATP detection reagent and by the enzyme that is not present in human body cell the component of ATP.Not only simple to operate, speed is fast compared with prior art for the method for the present invention, but also sensitivity and specificity with higher.
Description
Technical field
The present invention relates to there are the methods of mycoplasma pneumoniae in a kind of quickly detection clinical sample.
Technical background
Eaton agent pneumonia (MP) is caused by lower respiratory tract infection mycoplasma pneumoniae.Mycoplasma pneumoniae is a kind of cell-free
The bacteroid microorganism of the class of wall.Compared with Nosocomial, up to 20% Community acquired pneumonia is by pneumonia branch
Caused by substance.The common symptoms of mycoplasma pneumoniae infection include dry persistent coughs, duration low fever, sore throat, no
It is suitable, be slightly short of breath and pneumonia.In rare cases, infection can also result in arthritis, pericarditis, the nervous system disease,
Encephalitis, kidney failure, hemolytic anemia, skin disease and otopathy etc..
It has been known that there is more than 200 kinds of mycoplasma pneumoniaes can infect the mankind.It can be used for effectively controlling in spite of effective antibiotic
It treats, but effectively treatment and proper use of antibiotic need diagnosis promptly and accurately.Due to the clinical symptoms of mycoplasma pneumoniae infection
Similar to the infection of other pathogens, Accurate Diagnosis is difficult.For example, other bacteriums such as streptococcus and haemophilus and virus (packet
Include parainfluenza virus and influenza virus) infection symptom, it is closely similar with the clinical symptoms of mycoplasma pneumoniae infection, therefore very
It is easy to obscure with them.And use effective antibiotic treatment, it is necessary to dependent on promptly and accurately diagnosing whether be mycoplasma pneumoniae
Infection.
Currently, available mycoplasma pneumoniae infection test includes serology test and nucleic acid determination etc..Wherein, serology is examined
Mainly for strain, there are corresponding antibody (IgG and/or IgM) for survey.In addition, also it has been reported that developing similar thin for detecting
The rapid detection method of bacterium property antigen.But all there is disadvantage in all these detection methods.For example, former for pneumonia branch for detecting
The serology method of testing of the antibody of body cannot be used for the diagnosis of acute stage, because it is generally necessary to week age could can detect
Antibody；Antigen measuring usually lacks sensitivity again；Molecular assay operation based on nucleic acid is even more complicated and valuableness, makes it not
It is suitble to use in outpatient service.
Therefore, need a kind of method for capableing of accurate quick diagnosis mycoplasma pneumoniae infection, not only detect speed it is fast and
And it is easy to operate, it is easy to clinical use, and low in cost.
Summary of the invention
In view of defect existing for above-mentioned existing mycoplasma pneumoniae detection technique, the present invention provides a kind of clinical sample of quickly detection
There are the methods of mycoplasma pneumoniae in this, the antibody of mycoplasma pneumoniae neither in detection clinical sample, do not detect yet antigen or
Nucleic acid from mycoplasma pneumoniae, but detection is not present in human body cell but is present in the generation enzyme of the ATP in bovis cells
Activity, determine whether as mycoplasma pneumoniae infection.Specific technical solution is as follows:
There are the method for mycoplasma pneumoniae in a kind of quick detection clinical sample, comprise the following steps that
A) it reacts I: a certain amount of clinical sample being contacted with reagent I, the reagent I contains the ingredient for detecting ATP；
B) it reacts II: taking and contacted with the clinical sample of I moderate of reaction with reagent II, and the reagent II is in addition to containing
It is enough with react the identical group for ATP in test sample of ingredient in I exceptionally, also containing by being not present in human body
ATP generates the component that enzyme generates ATP；
C) incubate: the reaction mixture that will be prepared in step a) and step b) incubates one section at a proper temperature respectively
Time；
D) calculate signal ratio: respectively measurement reaction I and reaction II in ATP signal, and calculate reaction I signal with react II
In ATP signal ratio or net signal, by signal ratio or net signal come in judgement sample whether there is mycoplasma pneumoniae.
There are the methods of mycoplasma pneumoniae in quick detection clinical sample above-mentioned, further include sample elution step, i.e., will
The clinical sample carries out at elution before contacting with reagent I or reagent II by the sample buffer containing nonionic detergent
Reason.The sample includes the clinical sample collected from patient's throat, sputum or the lung from Diagnosis of Suspected Pneumonia mycoplasma infection patient
The sample of portion's acquisition.The concentration of nonionic detergent can cause the dissolution of cell membrane or perforation but not cause in the buffer
The dissolution or perforation of bacteria cell wall.
There are the method for mycoplasma pneumoniae in quick detection clinical sample above-mentioned, step a) and b) in, it is described for examining
The component for surveying ATP and the component for generating ATP are the reagent that biochemiluminescence can be generated based on luciferase.
As a preferred technical scheme, the component for detecting ATP include luciferase, fluorescein, magnesium ion,
DTT, coacetylase and buffer system appropriate.Preferably, the luciferase is firefly luciferase；Preferably, the magnesium from
Son selects magnesium acetate solution；Preferably, the buffer system is made of the HEPES of 0.5%BSA and pH=7.5.Further preferably
, the ultimate density of each substance is firefly luciferase 20mg/L, fluorescein 30mg/L, vinegar in the component for detecting ATP
Sour magnesium 10mM, DTT 5mM, coacetylase 0.5mM, buffer system 10mM.
As a preferred technical scheme, the ATP being not present in the human body generates enzyme and preferably includes acetokinase, ammonia
Any combination of base formate kinase, two kinases of pyruvate phosphate or three.
There are the method for mycoplasma pneumoniae in quick detection clinical sample above-mentioned, also wrapped in the reagent I and reagent II
It includes people's cell ATP and generates enzyme specific inhibitor.
There are the method for mycoplasma pneumoniae in quick detection clinical sample above-mentioned, in step c), the temperature of the incubation
It is 10~40 DEG C, the incubative time is less than 30min.
The beneficial effects of the present invention are:
The present invention passes through work that is being not present in detection people's cell but being present in the generation enzyme of the ATP in bovis cells
Property, to judge clinical sample with the presence or absence of mycoplasma pneumoniae.Not only speed is fast compared with prior art, but also with higher
Sensitivity and specificity.Meanwhile the method for the present invention is easy to operate, is easy to clinical use, and the excellent feature such as low in cost
It is suitble to outpatient service to use, quickly and effectively detects the presence of mycoplasma pneumoniae, whether quick patient diagnosed suffers from mycoplasma sense
Pneumonia is contaminated, treatment in time is conducive to, patient's benefited intensity is high.
The meaning respectively abridged:
ATP: atriphos
ADP: adenosine diphosphate (ADP)
AMP: adenosine monophosphate
DTT: dithiothreitol (DTT)
BSA: bovine serum albumin(BSA)
HEPES:4- (2- ethoxy) -1- piperazinyl ethanesulfonic acid
PPi: pyrophosphate
PPDK: two kinases of phosphoenolpyruvate
Specific embodiment
To make the object, technical solutions and advantages of the present invention clearer, below in conjunction with embodiment to skill of the invention
Art scheme is clearly and completely described.
Embodiment 1
Whether the present embodiment is whether to contain mycoplasma pneumoniae in determining clinical sample, to judge the patient by pneumonia branch
Pathogen infection.In the present embodiment, the clinical sample can be the sputum of the patient obtained by throat swab or from doubtful
The sample of lung's acquisition of mycoplasma pneumoniae infection patient.First by the clinical sample of acquirement in the sample containing nonionic detergent
It savors and suspends or elute in buffer, the concentration of nonionic detergent can cause the dissolution of cell membrane or wear in the sample buffer
Hole but not the dissolution or perforation for causing bacteria cell wall, thus can be special by the cellular membrane lysis of mycoplasma pneumoniae or wear
Hole, so that the ATP for releasing mycoplasma pneumoniae generates enzyme and ATP itself.
Since nonionic detergent simultaneously also can be by the cellular membrane lysis of human body cell in sample or perforation, to release
The ATP of human body.Therefore, two reaction groups are set up in the present embodiment to solve the problems, such as this.Reaction I: it is washed above-mentioned through nonionic
The clinical sample for washing agent processing is contacted with reagent I, and the reagent I contains the ingredient for detecting ATP；Reaction II: it takes and reacts I
The clinical sample of moderate is contacted with reagent II, in addition to containing enough groups for ATP in test sample in the reagent II
Exceptionally, also containing the component for generating ATP.The component of the component and generation ATP for detecting ATP is that can be based on fluorescence
Plain enzyme generates the reagent of biochemiluminescence, i.e. the present embodiment uses the ATP detection system based on luciferase, this system can produce
Third contact of a total solar or lunar eclipse signal is analyzed so that the later period measures.
In the present embodiment, the clinical sample after sample buffer of learning from else's experience respectively elution, is added to reagent I and reagent II
In, it is respectively formed II mixture of I mixture of reaction and reaction.Two reaction mixtures are both placed under common room temperature and are incubated, i.e.,
The condition of incubation is identical, and the time of incubation is usually less than 30min, then respectively measurement reaction I and reaction II in optical signal into
And judge in clinical sample with the presence or absence of mycoplasma pneumoniae.Concrete principle is as follows:
It shines in the reaction and needs three principle active components: luciferase, fluorescein and ATP.It is also required to magnesium simultaneously
Ion, DTT, coacetylase and buffer system appropriate.Therefore, the luminescence system based on firefly luciferase can be used for ATP's
Detection, and under the appropriate reaction conditions, the optical signal and ATP concentration correlation which generates.Therefore
Component described in reagent I for detecting ATP includes luciferase, fluorescein, magnesium ion, DTT, coacetylase and appropriate slow
Flushing system.It include acetokinase, carbamate kinase or pyruvate phosphate since the ATP present in mycoplasma pneumoniae generates enzyme
Two kinases etc., but these ATP generate enzyme and are not present in human body cell.Therefore in addition to including enough and reagent I in reagent II
Middle ingredient is identical for detecting each component of ATP, further comprises the component that these enzymes can be facilitated to generate ATP.In this way in situation,
The concentration (concentration of ATP existing for script) of background ATP in optical signal reflected sample in reaction I；Optical signal in reaction II
Background ATP concentration and newly-generated ATP concentration are then reflected, and newly-generated ATP concentration and ATP generate the activity and concentration of enzyme
It is positively correlated.In the present embodiment, the optical signal in reaction mixture is measured using luminometer, is to react by signal ratio
It is no to there is new ATP to generate, and then judge in clinical sample with the presence or absence of mycoplasma pneumoniae cell.Furthermore net signal (reacts II
Middle ATP signal subtracts in reaction I remaining signal after ATP signal) or signal multiple, it can also be used as the standard of judgement.And
Determine whether contain mycoplasma pneumoniae in sample, the cutoff value of signal ratio or net signal is by testing a large amount of clinical samples come really
Fixed, and determine that cutoff value is confirmed using generally acknowledged method such as nucleic acid detection method.
In some embodiments, the luciferase is preferably firefly luciferase；The magnesium ion preferably uses vinegar
Sour magnesium solution or magnesium chloride solution；The buffer system is preferably made of the HEPES of 0.5%BSA and pH=7.5.For detecting
The ultimate density of each substance is preferably firefly luciferase 20mg/L, fluorescein 30mg/L, magnesium acetate in the component of ATP
10mM, DTT 5mM, coacetylase 0.5mM, buffer system 10mM.
In the present embodiment, each solution formula is as follows:
Sample buffer (2X): BSA 0.5%, Triton X-100 1%, magnesium acetate 20mM, HEPES 100mM (pH tune
To 7.5).
(4X) ingredient: D- fluorescein 120mg/L, DTT 40mM, coacetylase 2mM, BSA is detected for ATP in reagent I
1%, firefly luciferase 80mg/L.
For facilitating ATP to generate the related component of enzyme production ATP in reagent II, it is specifically dependent upon and will test which kind of ATP life
At enzyme.
When for acetokinase detection, reagent II a (4X) are as follows: acetyl phosphate 12mM, ADP 12mM；The then reaction of the reaction
Path are as follows:
When for carbamate kinase detection, reagent II b (4X) are as follows: carbamyl phosphate 12mM, ADP 12mM；The reaction
Response path is as follows:
When detection for PPDK, reagent II c (4X) are as follows: phosphoenolpyruvate ester 12mM, ammonium chloride 100mM, AMP
12mM, pyrophosphate 12mM；The response path of the reaction is as follows:
When for detecting acetokinase and carbamate kinase simultaneously, reagent II d (4X) are as follows: acetyl phosphate 12mM, ammonia first
Acyl phosphatase 11 2mM, ADP 12mM；The response path of the reaction is as described in reagent II a and reagent II b.
For above-mentioned plurality of reagents II, how to select to carry out diagnosis of pneumonia mycoplasma infection using optimal reagent, and
It is determined by testing a large amount of clinical samples.For example, if the detection that mycoplasma ATP generates enzyme is confined to acetokinase,
Then reagent II is reagent II a；If the detection that mycoplasma ATP generates enzyme is confined to carbamate kinase, reagent II is
For reagent II b；If the detection that mycoplasma ATP generates enzyme is confined to PPDK, reagent II is reagent II c；If branch is former
When body ATP generates enzyme while detecting acetokinase and carbamate kinase, then reagent II is reagent II d, and is ground by clinic
Study carefully confirmation.Compared with traditional test, by whether fast containing mycoplasma speed in this method measurement clinical sample, and have
Higher sensitivity and specificity.
Embodiment 2
Because in a liquid state, in each reagent some main components should not be transported at room temperature with Long-term Cold Storage save, these
Ingredient can be lyophilized.Such as in certain embodiments, reagent I, freeze-dried reagent II can be freeze-dried, and buffer is the examination of 1X liquid
Agent.The buffer can be used for sample cleaning solution, and the buffer containing detection sample after washing is used directly for detecting.
In one embodiment, three kinds of agent prescriptions are as follows:
Sample buffer (1X): BSA 0.5%, Triton X-100 0.5%, magnesium chloride 10mM, HEPES 50mM (pH
It is adjusted to 7.5).
It is as follows that reagent I (detects) 4X formula for ATP: D- fluorescein 120mg/L, DTT 40mM, coacetylase 2mM, BSA
1%, firefly luciferase 80mg/L；62.5 μ l are taken, are lyophilized in a test tube.
Reagent II (for acetokinase and carbamate kinase ATP production and ATP detection, 4X) formula is as follows: acetyl phosphorus
Sour 12mM, carbamyl phosphate 12mM, ADP 12mM, D- fluorescein 120mg/L, DTT 40mM, coacetylase 2mM, BSA 1%, firefly
Fireworm luciferase 80mg/L；62.5 μ l are taken, are lyophilized in a test tube.
In above-described embodiment, reagent I and reagent II are lyophilized into single part detection reagent, thus simplify detection process.Inspection
Survey process is as follows: clinical sample being washed to suspension in buffer, takes the buffer 0.25ml containing sample that reagent I is added respectively
In reagent II, a period of time is incubated, is placed in photometer and detects optical signal, calculates optical signal ratio.
The optium concentration of the component of reagent I and reagent II determines after needing to test by a large number of experiments.Similarly, detector bar
Part such as temperature and detection time are also required to determine after a large number of experiments is tested.These tests can be by using a large amount of negative pharynxs
The enzyme of swab sample and standard items such as recombinant expression, such as acetokinase and carbamate kinase.Bacterium includes the acetic acid of mycoplasma
The nucleic acid sequence of kinases and carbamate kinase can be found delivering to reach in document.The gene of these enzymes can be by manually closing
It at forming, is then implanted in coli expression system, obtains these enzymes to separate in Escherichia coli.And these recombinations
Enzyme can be used for Optimum Experiment and Quality Control.
In the present embodiment, find that acetokinase and carbamate kinase provide the optimal of enough sensitivity through overtesting
Or close to the concentration of component of optimal reagent I and reagent II be above-mentioned 4x concentration, i.e., the 62.5 μ l 4x concentration are taken respectively
Reagent I and reagent II freeze-drying are with detection reagent pipe, and taking 0.25ml sample, (a small amount of enzyme is added to the simulation after above-mentioned buffer
Sample) it is added in reagent I and II, reach enough signal ratios after incubation at room temperature 15 minutes.Experiments have shown that incubation temperature
It is 10~40 DEG C, incubative time is no more than 30mim, and optimal incubation temperature is 37 DEG C, and optimal incubative time is that 30mim can be obtained
Better signal ratio, but in view of clinical use is convenient, in the present embodiment, ambient temperature is still used, incubates 15min.
3 comparative example of embodiment
After the component and concentration, dosage form and checking procedure and condition of reagent determine, it is critical that test determination can be carried out
It is worth (or cutoff value).Confirm with 100 by detection of nucleic acids for example, being confirmed as feminine gender by detection of nucleic acids by test 100
For positive sample.By the test to this 200 clinical samples determine critical value (or cutoff value) be 2.15 when sensitivity and
Specificity be it is best, respectively 93% and 95%.In the present embodiment, detection of nucleic acids is assumed to compare reagent (i.e. goldstandard side
Method).
A clinical test is carried out based on the critical value (2.15) of above-mentioned determination.In this embodiment, clinical test has 3
Clinical test point, sharing 1000 has the patient of mycoplasma infection symptom to participate in clinical test.Every participant takes two pharynxs to wipe
Son, for detection of nucleic acids diagnosis (such as fluorescence PCR method), another is detected for the method for the present invention for one of them.
In the present embodiment, testing result is as follows:
1. the method for the present invention nucleic acid detection method sensitivity and specificity of table compare
According to table 1, compared with nucleic acid detection method, the sensitivity and specificity of the method for the present invention are respectively
92.77% and 97.43%.
4 comparative example of embodiment
Since penicillin class antibiotic (most of antibiotic is such antibiotic) and antiviral class drug are to treatment
Mycoplasma infection is invalid, therefore in order to suit the remedy to the case, and quick and precisely detects mycoplasma infection to treatment mycoplasma infection to Guan Chong
It wants.In addition, being used in outpatient clinic detection, simple operations step, miniature instrument, and economic reagent is also critically important.Separately
Outside, optimum sampling and detection time are acute stage, therefore the sensitivity of acute stage detection is also critically important.Following table lists several normal
With the comparison of detection reagent.
Mycoplasma pneumoniae is often compared with detection method in 2. clinic of table
As seen from Table 2, compared with traditional ELISA method, colloidal gold chromatography and conventional fluorescent PCR method, this hair
The bright method detection used time is only 10-20 minutes, is maintained an equal level with the famous colloidal gold chromatography of speed, and much due to ELISA
Method and conventional fluorescent PCR method；Secondly, equally compared with nucleic acid detection method, the sensitivity and specificity of the method for the present invention detection
Reach 90% or more, this is much superior to ELISA method, colloidal gold chromatography, and than conventional fluorescent PCR detection of nucleic acids
Method is not susceptible to sample contamination influence；In addition to this, the method for the present invention also have cost of labor is low, instrument cost is low, reagent at
This is low, easy to operate, outpatient service is suitble to the excellent feature such as to use, and quickly and effectively detects whether patient suffers from mycoplasma sense
Pneumonia is contaminated, treatment in time is conducive to, patient's benefited intensity is high.
It is obvious to a person skilled in the art that invention is not limited to the details of the above exemplary embodiments, Er Qie
In the case where without departing substantially from spirit or essential attributes of the invention, the present invention can be realized in other specific forms.Therefore, no matter
From the point of view of which point, the present embodiments are to be considered as illustrative and not restrictive.Although in addition, it should be understood that originally
Specification is described in terms of embodiments, but only includes not one technical solution, and this narrating mode of specification is only
It is only for clarity that the skilled in the art should refer to the specification as a whole, the technical solution in embodiment can also
To be properly combined, form other embodiments that can be understood by those skilled in the art.
Claims (10)
1. in a kind of quickly detection clinical sample, there are the methods of mycoplasma pneumoniae, it is characterised in that: comprises the following steps that
A) it reacts I: a certain amount of clinical sample being contacted with reagent I, the reagent I contains the component for detecting ATP；
B) it reacts II: taking and contacted with the clinical sample of I moderate of reaction with reagent II, and the reagent II is in addition to containing enough
With react the identical group for ATP in test sample of ingredient in I exceptionally, also containing can be by being not present in human body
ATP generates the component that enzyme generates ATP；
C) incubate: the reaction mixture that will be prepared in step a) and step b) is incubated under identical incubation conditions respectively
For a period of time；
D) calculate signal ratio: measuring the light letter for being able to reflect ATP concentration in the reaction I after step c) incubates and reaction II respectively
Number, and the ratio or net signal for reacting I signal and reacting II signal are calculated, and by signal ratio or net signal come in judgement sample
With the presence or absence of mycoplasma pneumoniae.
2. in quick detection clinical sample according to claim 1, there are the methods of mycoplasma pneumoniae, it is characterised in that: also
Including sample elution step, i.e., the clinical sample through before contacting with reagent I or reagent II and containing nonionic detergent
Sample buffer carry out elution processing, the concentration of the nonionic detergent can cause cell membrane dissolution or perforation but not
Cause the dissolution or perforation of bacteria cell wall.
3. feature exists there are the method for mycoplasma pneumoniae in quick detection clinical sample according to claim 1 or 2
In: the clinical sample include the sample collected from patient's throat, sputum or lung from Diagnosis of Suspected Pneumonia mycoplasma infection patient
The sample of portion's acquisition.
4. in quick detection clinical sample according to claim 1, there are the methods of mycoplasma pneumoniae, it is characterised in that: step
It is rapid a), b) in, it is described for detect ATP component and generate ATP component be can based on luciferase generate bioid
Learn luminous reagent.
5. feature exists there are the method for mycoplasma pneumoniae in quick detection clinical sample according to claim 1 or 4
In: step a), b) in, the component for detecting ATP includes that luciferase, fluorescein, magnesium ion, DTT, coacetylase are gentle
Flushing system.
6. in quick detection clinical sample according to claim 5, there are the methods of mycoplasma pneumoniae, it is characterised in that: institute
Stating luciferase is firefly luciferase；It is magnesium acetate solution or magnesium chloride solution that the magnesium ion, which is selected,；The buffer system
System is made of the HEPES of 0.5%BSA and pH=7.5.
7. feature exists there are the method for mycoplasma pneumoniae in quick detection clinical sample according to claim 5 or 6
In: the ultimate density of each substance is firefly luciferase 20mg/L, fluorescein 30mg/ in the component for detecting ATP
L, magnesium ion solution 10mM, DTT 5mM, coacetylase 0.5mM, buffer system 10mM.
8. in quick detection clinical sample according to claim 1, there are the methods of mycoplasma pneumoniae, it is characterised in that: institute
Stating the ATP being not present in human body and generating enzyme includes acetokinase, carbamate kinase, two kinases of pyruvate phosphate or three
Any combination.
9. in quick detection clinical sample according to claim 1, there are the methods of mycoplasma pneumoniae, it is characterised in that: institute
State the specific inhibitor for generating enzyme in reagent I and reagent II containing people's cell ATP.
10. in quick detection clinical sample according to claim 1, there are the methods of mycoplasma pneumoniae, it is characterised in that:
In step c), the temperature of the incubation is 10~40 DEG C, and the incubative time is less than 30min.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20030049708A1 (en) * | 1999-11-08 | 2003-03-13 | Lumitech (Uk) Limited | Assay of micro-organisms in cell cultures |
| CN1756848A (en) * | 2003-04-17 | 2006-04-05 | 坎布里克斯生物科学诺丁汉有限公司 | Assay for detecting mycoplasma by measuring acetate kinase or carbamate kinase activity |
-
2019
- 2019-07-22 CN CN201910660493.4A patent/CN110283882A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20030049708A1 (en) * | 1999-11-08 | 2003-03-13 | Lumitech (Uk) Limited | Assay of micro-organisms in cell cultures |
| CN1756848A (en) * | 2003-04-17 | 2006-04-05 | 坎布里克斯生物科学诺丁汉有限公司 | Assay for detecting mycoplasma by measuring acetate kinase or carbamate kinase activity |
Non-Patent Citations (1)
| Title |
|---|
| VAHID MOLLA KAZEMIHA, ET AL.: "Sensitivity of biochemical test in comparison with other methods for the detection of mycoplasma contamination in human and animal cell lines stored in the National Cell Bank of Iran", 《CYTOTECHNOLOGY》 * |
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