CN1990876A - Staphylococcus drug sensitive batten and preparation method thereof - Google Patents

Abstract

The invention discloses an aureus drug sensitivity lath. The antibiotic on lath is prepared according to clinical requirement and regulation, and anti- oxidation agent is added into it to prolong its storage period. The product can direct clinical medication and is very great value of clinical application.

Description

Staphylococcus drug sensitive batten and preparation method thereof

Technical field:

The invention belongs to the biologic product technology field.Be specifically related to a kind of staphylococcus (MS) drug sensitive batten and preparation method thereof.

Background technology:

Over nearly 20 years, owing to extensively use miscellaneous antibacterials in large quantities clinically, drug-resistance of bacteria is more and more general, and the bacterium that has is anti-2-7 kind medicine simultaneously, and the wide-scale distribution of Resistant strain is brought very big difficulty to clinical treatment.The clinical diagnosis and treatment means are brought in constant renewal in addition, though improved diagnosis and result of treatment, have also brought more cross infection chance, make modulation on immune status be tending towards low.Especially noticeable is that very big transition have taken place infection type, and be among the variation, in order to adapt to this new variation, the research fresh condition, solve the new problem that constantly occurs, formed a new subject gradually--clinical microbiology (Clinical Microbiology).

In the last few years, hospital infection (Nosocomial infection) more and more becomes one of problem that people extremely pay close attention to.So-called hospital infection is meant that patient occurred in the while in hospital and the just infection of morbidity soon of leaving hospital, but does not comprise that patient has begun before being admitted to hospital or be in infection in latent period when being admitted to hospital.So the infection of the characteristics of hospital infection in appearing at hospital environment, and take place popular, so can be referred to as hospital acquired infections (Hospital acquired infection) again.Nineteen eighty-three, 11, the 295 routine patients of 21 hospitals, the incidence average out to 8.4% of hospital infection were investigated by the Ministry of Health.Though the state of an illness of generation hospital infection has and gently has heavily, has all increased extra misery, economical load and misfortune to patient, also can increase the weight of the burden of medical treatment and nursery work simultaneously owing to the prolongation hospital stays, influence sick bed and have enough to meet the need.The disease that serious hospital infection often makes patient is suffered from can not reach the curative effect of expection, even produces sequela or the death that is difficult to treat.According to an incomplete statistics data report of the U.S., have 5% hospital infection takes place among its inpatient approximately, the average 4～5d that is in hospital that prolongs has 100,000 people's death every year approximately in hospital infection, estimate annual since hospital infection to consume 20 more surplus hundred million dollars.

From causing the microorganism of hospital infection, no doubt some is still the pathogenic very strong microorganism that we are familiar with, they are everlasting and cause that complication infects after the some diseases, but the pathogenic agent kind of hospital infection in recent years has the trend of considerable change, it was the gram-positive cocci of main pathogen originally that Gram-negative bacteria has replaced, wherein staphylococcus section and pseudomonas account for the 60%-65% of hospital infection, and are mainly many drug tolerant bacterias.Wherein major part is human body normal microflora, fungi and virus, and also there are some reports the parasite aspect.

With regard to entire society, hospital is a susceptible host, because the patient of hypoimmunity concentrates (comprising infant and the elderly), the continual renovation of diagnostic means (as the application of endoscope, intubate, conduit etc.), though improved diagnosis and result of treatment, also increased the chance of cross infection.Owing to use microbiotic widely, the continuous transition of bacterium have encouraged the existence and the development of nosocomial infection, and the clinical microbiology most important character of today is to change to the hospital infection aspect.

Microorganism identification and susceptibility analytical system are the high-tech in-vitro diagnosis products that is mainly used in clinical labororatory, and be significant to auxiliary clinical each section office's diagnoses and treatment.At present in the world, Rapid identification and the susceptibility analysis of bacterium obtained very big progress, microorganism automatization testing product has progressively substituted manual inspection.

At present, like product mainly contains in the world: the MicroScan automatic microbe of the VITEK-AMS microbiological analysis system of French biology-Mei Liai company, U.S. Dade-MicroScan company is identified and the SENSTITRE fluorescent method fast microbiological of antibiotics susceptibility test system and Britain Trek Diagnostic System Ltd. is identified and the susceptibility analytical system.Their principle of work is close, mainly be difference according to the bacterium physico-chemical property, adopt photoelectric colorimetry (susceptibility is a turbidimetry), measure bacterial growth and decompose substrate and cause pH value to change and the different colours that produces and turbidity change and judge response situation, thereby provide the identification and susceptibility result.

At present, for adapting to the demand of specialty of clinical labororatory to microorganism identification susceptibility equipment, have higher requirement to the function of microflora product in market, the microniological proudcts that development and operation is easy, the evaluation scope is wide, susceptibility MIC value is accurately practical, also can provide experimental result have fast become a kind of an urgent demand.

Though the microbial identification system of BIOLOG company is the Identifying micro-organisms bacterial classification comparatively exactly, but lacking susceptibility (microbiotic MIC) detects, and when doing the clinical expansion of Micro biological Tests product, the general system that all requires possesses two kinds of functions of MIC (minimal inhibitory concentration) value of identifying bacterial classification and detect antibiotics simultaneously, to instruct clinical application.For meeting the need of market, and in competition, occupy advantage.

And the drug sensitive batten kind of product is less now, can not satisfy the drug sensitive test of all kinds bacterium, antibiotic concentration gradient is less in the lath in addition, and major part is reported sensitivity, medium sensitivity and resistance result qualitatively only, and can accurately not report out quantitative MIC value.

Summary of the invention:

Technical problem to be solved by this invention is to overcome above-mentioned weak point, a kind of susceptibility fast and accurately of research and design detection system.

The invention provides a kind of staphylococcus (MS) drug sensitive batten.

Susceptibility of the present invention detects to be tested based on following principle:

Linear relationship according to each medicine growth slope and MIC (minimal inhibitory concentration), select the extent of dilution of proper number, the bacteria suspension that adds bacterium to be checked, after 4-15h is hatched, use photoelectricity than turbid principle, can obtain the slope of bacterium to be checked in each concentration, compare with positive control hole deviation rate, and calculate the compound slope of bacterium to be checked, obtain the MIC value through regression analysis, and obtain the result of corresponding sensitivity (S), medium sensitivity (I) and resistance (R) according to the NCCLS standard.

On the other hand, the inventor improves the stability of drug sensitive batten through great deal of experimental.Because in drug sensitive batten of the present invention, added microbiotic, because oxidation very easily takes place the performance of microbiotic own, just can lose efficacy in the short period of time, storage has caused very big influence to drug sensitive batten.Therefore the present invention selects for use antioxidant to add the preservation period that prolongs drug sensitive batten in the drug sensitive batten.

Antioxidant (AOA) also be scavenging agent, be startup and hyperplasia (spreading) process that a class can suppress or block free chain reaction, reduce number of free radical, delay the old process of sorrow, the compound antioxidant and the free radical that improve vitality are a pair of paradox.The molecular structure of antioxidant is more special, and its electronics is single arrangement, and it can provide in the electronics and free radical, and self can not be formed with the material of harm, and chain reaction can not take place yet, and reaches oxidation resistant purpose by removing free radical.How antioxidant works

1. normal Sauerstoffatom has 4 pairs of electronics, and the body eubolism can make atom lose an electronics, has so just formed free radical, and free radical mainly makes oneself pairing by the electronics of grabbing on other his molecule.

2. when free radical is captured an electronics from cytolemma after, just produced another new free radical, and begun chain reaction.

3. electronics is captured chain reaction erosion cytolemma, and the forfeiture that causes the cell integrity is that cancer and other disease have been opened convenience.

4. because its specific molecule structure, antioxidant can provide that an electronics is given free radical and self can not form the deleterious Hazardous substances that can cause chain reaction.

Common antioxidant has VC, VE, selenium, germanium, veronal, glycine, sodium bisulfite, tea-polyphenol class, spice extract.

Amino acid and dipeptide amino acid such as methionine(Met), tryptophane, phenylalanine, proline(Pro) etc. can both with metal ion-chelant, so their good auxiliary antioxidants.In recent years, the food scientist finds that the L-Ala end is that 9 kinds of dipeptides of nitrogen are all stronger than any amino acid whose resistance of oxidation.Wherein the strongest with L-Ala-Histidine, L-Ala-tyrosine, 3 kinds of dipeptides resistance of oxidation of L-Ala-tryptophane especially, be worth exploitation energetically.

Spice extract is as far back as the thirties in 20th century, and people study the antioxygenation of spice with regard to beginning.To the fifties, the scientific research personnel analyzes 32 kinds of spices, find antioxidant property wherein best be Rosmarinus officinalis and Salvia japonica Thunb..This series products contains multiple antioxidant components such as flavonoid, terpenoid, organic acid more, can cut off greasy autoxidation chain, chelated metal ions, and play and the organic acid synergistic function.Rosmarinus officinalis (Rosmarinus officinalisL.) is a Labiatae, belongs to evergreen perennial filling and wants the incense drug mould thing of tool high anti-oxidation ability.Its resistance of oxidation comes from the aldehydes matter (phenolics) that includes, serve as main wherein with phenolic diterpenes, as Salvin (camosicacid), carnosol (carnosol), rosmarinic acid rosemary extract Nantural non-toxics such as (rosmarinic acid), anti-oxidation efficacy is higher than natural antioxidantss such as existing VC, VE, tea-polyphenol far away, resistance of oxidation than the oxygenant BHT of synthetic and BHA is strong more than 4 times, and its Stability Analysis of Structures, is difficult for decomposing.Therefore rosemary antioxidant is a natural antioxidant of new generation, has thoroughly avoided the toxic side effect of synthetized oxidation preventive agent and the characteristics of pyrolytic decomposition.

The inventor selects the use of antioxidant by following test:

1, experimental technique

Adopted veronal, glycine and Rosmarinus officinalis to be configured to the aqueous solution and the aqueous solution that does not add antioxidant respectively and added that microbiotic is mid-to become drug sensitive batten, stored for-10 ℃ as antioxidant.Selection standard bacterial strain 29213 adds drug sensitive batten to be tested, and the microbiotic MIC value of testing 4 kinds of laths for the first time is all in specialized range.First month carried out one-time detection weekly, from per two months per two weeks carrying out one-time detection, carries out one-time detection after six months every other month, to all losing efficacy to all microbiotic.Every kind of lath of each test is tested ten times altogether.

2, lath working method (clinical setting during use)

(1), get 27853 usefulness reason salt solution and dispose 0.5 Maxwell bacteria suspension, bacteria suspension and cultured solution of broth are diluted (being that every 12ml meat soup adds the 60ul bacteria suspension) by 1: 200 times.

(2), the bacterium liquid that configures is added in the drug sensitive batten with pipettor, every hole adds 100ul.

(3), lath being placed 35 ℃ of incubators cultivated 24 hours.

(4), read the lath result, if it is long into dirt to become turbid in the hole, as if haze-free for not growing, every kind of first concentration value of not growing the hole of microbiotic is correct MIC value.

3, experimental result

This experiment has been carried out 1 year altogether, and experimental result is as shown in the table:

The microbiotic title	The antioxidant service condition			Do not add antioxidant
					Veronal	Glycine	Rosmarinus officinalis
	Ciprofloxacin	Lost efficacy in six months	Lost efficacy in six months	Lost efficacy after 1 year					Lost efficacy in nine months
Ofloxacine USP 23					Lost efficacy in six months	Lost efficacy in six months	Lost efficacy in ten months	Lost efficacy in seven months
	Gentamicin	Lost efficacy in two months	Lost efficacy in two months	Lost efficacy in ten months					Lost efficacy in seven months
Amikacin					Lost efficacy in two months	Lost efficacy in two months	Lost efficacy in 11 months	Lost efficacy in seven months
	Tobramycin	Lost efficacy in six months	Lost efficacy in six months	Lost efficacy after 1 year					Lost efficacy in nine months
Piperacillin					Lost efficacy in two months	Lost efficacy in two months	Lost efficacy after 1 year	Lost efficacy in nine months
	Aztreonam	Lost efficacy in six months	Lost efficacy in six months	Lost efficacy after 1 year					Lost efficacy in nine months
Imipenum					Lost efficacy in two months	Lost efficacy in two months	Lost efficacy in ten months	Lost efficacy in seven months
	Ceftazime	Lost efficacy in six months	Lost efficacy in six months	Lost efficacy after 1 year					Lost efficacy in nine months
Cefoperazone					Lost efficacy in six months	Lost efficacy in six months	Lost efficacy in 11 months	Lost efficacy in nine months
	Ceftriaxone	Lost efficacy in six months	Lost efficacy in six months	Lost efficacy after 1 year					Lost efficacy in nine months
Cefepime					Lost efficacy in six months	Lost efficacy in six months	Lost efficacy after 1 year	Lost efficacy in nine months

4, interpretation of result

Test-results shows:

(1), use veronal, glycine antioxidant, inefficacy has just appearred in lath some microbiotic after two months, shows that these two kinds of microbiotic not only prolong the preservation period of susceptibility, can influence antibiotic performance on the contrary, has shortened the period of storage of lath.So two kinds of antioxidants can not be selected for use.

(2), use the rosemary antioxidant microbiotic just to have some microbiotic to occur losing efficacy later at ten months, obviously long than the lath shelf time that does not add antioxidant.So using Rosmarinus officinalis is feasible as the preservation period that antioxidant adding drug sensitive batten prolongs drug sensitive batten.

On the basis of above-mentioned test, the present invention has selected rosemary antioxidant for use, strengthens the stability of drug sensitive batten, prolongs the preservation period of drug sensitive batten.

Another object of the present invention has provided the preparation method of staphylococcus drug sensitive batten, and this method comprises the following steps:

(1) configuration microbiotic diluent

It is stand-by as antibiotic diluent to add (Alecrim antioxidant) concentration in the distilled water and be 100,000 of distilled water/to 1,000,000/be configured to the aqueous solution;

(2) packing

It is stand-by that the diluent that configures is packed as every test tube 15ml;

(3) configuration stoste

Require the various antibiotic initial stostes of configuration according to dilution;

(4) packing liquid material

According to the requirement of various lath concentration dilution, the microbiotic stoste of step (3) is joined in step (2) test tube that corresponding branch installs.

(5) application of sample packing lath

Use to divide assembling system in the corresponding position in 96 holes to specified drug sensitive batten application of sample, every hole adds 50ul;

(6) lath drying

Above-mentioned lath vacuum-drying 16-18 hour;

(7) packing

Above-mentioned antibiotic concentration dilution requires:

The microbiotic title	The position	The solution title	Consumption (ul)
				No. 17 medicine Ciprofloxacins	A1	--	--
B1	--	--
			C1		No. 17 medicine stostes	180
D1	No. 17 medicine stostes	90
			E1		No. 17 medicine stostes	45
F1	No. 17 medicine diluents	225
			G1		No. 17 medicine diluents	112
H1	No. 17 medicine diluents	56

The microbiotic title	The position	The solution title	Consumption (ul)
				No. 3 medicine amikacins	A2	No. 3 medicine stostes	288
B2	No. 3 medicine stostes	144
			C2		No. 3 medicine stostes	72
D2	No. 3 medicine stostes	36
			E2		No. 3 medicine diluents	180
F2	No. 3 medicine diluents	90
			G2		No. 3 medicine diluents	45
H2	No. 3 medicine diluents	23

The microbiotic title	The position	The solution title	Consumption (ul)
				No. 16 medicine Rifampins	A3	No. 16 medicine stostes	480
B3	No. 16 medicine stostes	240
			C3		No. 16 medicine stostes	120
D3	No. 16 medicine stostes	60
			E3		No. 16 medicine stostes	30
F3	No. 16 medicine diluents	150
			G3		No. 16 medicine diluents	75
H3	No. 16 medicine diluents	38

The microbiotic title	The position	The solution title	Consumption (ul)
				No. 14 medicine erythromycin	A4	No. 14 medicine stostes	384
B4	No. 14 medicine stostes	192
			C4		No. 14 medicine stostes	96
D4	No. 14 medicine stostes	48
			E4		No. 14 medicine diluents	240
F4	No. 14 medicine diluents	120
			G4		No. 14 medicine diluents	60
H4	No. 14 medicine diluents	30

The microbiotic title	The position	The solution title	Consumption (ul)
				No. 12 medicine clindamycins	A5	No. 12 medicine stostes	384
B5	No. 12 medicine stostes	192
			C5		No. 12 medicine stostes	96
D5	No. 12 medicine stostes	48
			E5		No. 12 medicine diluents	240

No. 12 medicine clindamycins	F5	No. 12 medicine diluents	120
				G5	No. 12 medicine diluents	60
	H5	No. 12 medicine diluents	30

The microbiotic title	The position	The solution title	Consumption (ul)
				No. 11 medicine SMZ	A6	No. 11 medicine stostes	684
B6	No. 11 medicine stostes	342
			C6		No. 11 medicine stostes	171
D6	No. 11 medicine stostes	86
			E6		No. 11 medicine stostes	42
F6	No. 11 medicine diluents	210
			G6		No. 11 medicine diluents	105
H6	No. 11 medicine diluents	53

The microbiotic title	The position	The solution title	Consumption (ul)
				No. 15 medicine vancomycins	A7	No. 15 medicine stostes	360
B7	No. 15 medicine stostes	180
			C7		No. 15 medicine stostes	90
D7	No. 15 medicine stostes	45
			E7		No. 15 medicine diluents	225
F7	No. 15 medicine diluents	112
			G7		No. 15 medicine diluents	56
H7	No. 15 medicine diluents	28

The microbiotic title	The position	The solution title	Consumption (ul)
				No. 19 medicine penicillin	A8	No. 19 medicine stostes	45
B8	No. 19 medicine diluents	225
			C8		No. 19 medicine diluents	112
D8	No. 19 medicine diluents	56
			E8		No. 19 medicine redilution liquid	280
F8	No. 19 medicine redilution liquid	140
			G8		No. 19 medicine redilution liquid	70
H8	No. 19 medicine redilution liquid	35

The microbiotic title	The position	The solution title	Consumption (ul)
				No. 13 medicine Oxazacillins	A9	No. 13 medicine stostes	240
B9	No. 13 medicine stostes	120
			C9		No. 13 medicine stostes	60
D9	No. 13 medicine stostes	30
			E9		No. 13 medicine diluents	150
F9	No. 13 medicine diluents	75
			G9		No. 13 medicine diluents	38
H9	No. 13 medicine diluents	19

The microbiotic title	The position	The solution title	Consumption (ul)
				No. 6 medicine Kefzols	A10	No. 6 medicine stostes	288
B10	No. 6 medicine stostes	144
			C10		No. 6 medicine stostes	72
D10	No. 6 medicine stostes	36
			E10		No. 6 medicine diluents	180
F10	No. 6 medicine diluents	90
			G10		No. 6 medicine diluents	45
H10	No. 6 medicine diluents	23

The microbiotic title	The position	The solution title	Consumption (ul)
				No. 41 medicine cefaclors	A11	No. 41 medicine stostes	288
B11	No. 41 medicine stostes	144
			C11		No. 41 medicine stostes	72
D11	No. 41 medicine stostes	36
			E11		No. 41 medicine diluents	180
F11	No. 41 medicine diluents	90
			G11		No. 41 medicine diluents	45
H11	No. 41 medicine diluents	23

The microbiotic title	The position	The solution title	Consumption (ul)
				No. 10 medicine ceftriaxones	A12	No. 10 medicine stostes	288
B12	No. 10 medicine stostes	144
			C12		No. 10 medicine stostes	72
D12	No. 10 medicine stostes	36
			E12		No. 10 medicine diluents	180
F12	No. 10 medicine diluents	90
			G12		No. 10 medicine diluents	45
H12	No. 10 medicine diluents	23

MS lath susceptibility concentration gradient table

Annotate: be the scope between medicaments insensitive and resistance between the shade.(with reference to the NCCLS2004 standard)

Product of the present invention is compared with existing Micro biological Tests product, has more the clinical application directive significance, can reduce growing of Resistant strain, and reduces various infection in the institute.Statistical analysis system can be according to different statistical condition, in one period, analysis on the epidemiology is made in each lesion of hospital, this crowd zone and surrounding area, and the effect of monitoring is played in the control of the variation of special Resistant strain and disease, bigger clinical value is arranged.

Embodiment:

Example:

The MS staphylococcus drug sensitive batten is an example, and the technological process of production of producing 250 products in batches is as follows:

1, the configuration Rosmarinus officinalis aqueous solution

Taking by weighing 0.03 gram Rosmarinus officinalis (Rosmarinus officinalis ultimate density be 100,000/) fully is dissolved in the 1500ml deionized water.

Or take by weighing 0.3 gram Rosmarinus officinalis (Rosmarinus officinalis ultimate density be ten thousand/) and fully be dissolved in the 1500ml deionized water.

Or take by weighing 0.003 gram Rosmarinus officinalis (Rosmarinus officinalis ultimate density be 1,000,000/) and fully be dissolved in the 1500ml deionized water.

2,, evenly divide then to install in 96 test tubes that disinfect every pipe packing 15ml with the solution autoclaving that configures.

3, dispose the initial liquid of each microbiotic.

Concentration according to following table disposes each antibiotic initial liquid:

Medicine name	The medicine numbering	Initial liquid concentration
			Ciprofloxacin	17	0.00133gl/ml
Amikacin	3	0.0133g/ml
			Rifampin	16	0.002g/ml
Erythromycin	14	0.00125g/ml
			Clindamycin	12	0.00125g/ml
SMZ	11	0.0133g/ml
			Vancomycin	15	0.00533g/ml
Penicillin	19	0.00133g/ml
			Prostaphlin	13	0.002g/ml
Kefzol	6	0.00667g/ml
			Cefaclor	41	0.00667g/ml
Ceftriaxone	10	0.0133g/ml

4, dispose each hole antidusting cellulose solution

According to following table the plain concentration dilution table of antidusting add in the stand-by 15ML test tube of 96 packing of lath by the volume that shows in the table.

Staphylococcus lath (No. 2 plates) MS
												The medicine numbering	The medicine name	Sequence number	A	B	C	D	E	F	G	H	Total fluid volume
17	Ciprofloxacin	1	Blank	Positive	180	90	45	225	112	56	415
												3	Amikacin	2	288	144	72	36	180	90	45	23	640
16	Rifampin	3	480	240	120	60	30	150	75	38	1030
												14	Erythromycin	4	384	192	96	48	240	120	60	30	820
12	Clindamycin	5	384	192	96	48	240	120	60	30	862
												11	SMZ	6	684	342	171	86	42	210	105	53	1383
15	Vancomycin	7	360	180	90	45	225	112	56	28	775
												19	Penicillin	8	45	225	112	56	280	140	70	35	145
13	Prostaphlin	9	240	120	60	30	150	75	38	19	550
												6	Kefzol	10	288	144	72	36	180	90	45	23	640
41	Cefaclor	11	288	144	72	36	180	90	45	23	640
												10	Ceftriaxone	12	288	144	72	36	180	90	45	23	640

Remarks:

1. above unit all is ul

2.. boldface type is represented with 10 times of stoste dilutions: i.e. 100ul stoste+900ul water diluent.

3. italics is represented with 10 times of diluent redilution: i.e. 100ul diluent liquid+900ul water

5, application of sample packing lath

96 cuvette cartridges that configure are being added on the model machine, using to add model machine to specified drug sensitive batten application of sample, every hole adds 50ul.

(to move the branch assembling system earlier before this; Whole procedure approximately needs 3 hours).

6, lath drying

The vacuum pump that the lath that application of sample is good is put into drying room carries out vacuum to be drained, and lath can drying finish after 24 hours.

7, lath packing

The lath that drying is good is delivered to make-up room from drying room through pass-through and is packed.

8, lath storage

The lath that packing finishes is put into-20 ℃ of refrigerators and is carried out freezing preservation.

Claims (4)

1, a kind of staphylococcus drug sensitive batten is characterized in that what this staphylococcus drug sensitive batten was made by following method:

(1) disposes antibiotic diluent

Add rosemary antioxidant in the distilled water, the concentration of adding be 100,000 of distilled water/to 1,000,000/, be configured to the aqueous solution, stand-by as antibiotic diluent;

(2) packing

It is stand-by that the diluent that configures is packed as every test tube 15ml;

(3) configuration stoste

Require the various antibiotic stostes of configuration according to dilution;

(4) packing liquid material

According to the requirement of lath concentration dilution, the microbiotic stoste of step (3) is joined in step (2) test tube that corresponding branch installs;

(5) application of sample packing lath

Use to divide assembling system in the corresponding position in 96 holes to specified drug sensitive batten application of sample, every hole adds 50ul;

(6) lath drying

Above-mentioned lath vacuum-drying 16-18 hour;

(7) packing.

2, a kind of preparation method of staphylococcus drug sensitive batten as claimed in claim 1 is characterized in that this method comprises the following steps:

(1) disposes antibiotic diluent

Add rosemary antioxidant in the distilled water, the concentration of adding be 100,000 of distilled water/to 1,000,000/, be configured to the aqueous solution, stand-by as antibiotic diluent;

(2) packing

It is stand-by that the diluent that configures is packed as every test tube 15ml;

(3) configuration stoste

Require the various antibiotic stostes of configuration according to dilution;

(4) packing liquid material

According to the requirement of lath concentration dilution, the microbiotic stoste of step (2) is joined in step (2) test tube that corresponding branch installs;

(5) application of sample packing lath

Use to divide assembling system in the corresponding position in 96 holes to specified drug sensitive batten application of sample, every hole adds 50ul;

(6) lath drying

Above-mentioned lath vacuum-drying 16-18 hour;

(7) packing.

3, the preparation method of staphylococcus drug sensitive batten according to claim 2 is characterized in that in wherein said step (4) packing liquid material and (5) the application of sample packing lath, stoste consumption and as follows at the branch holding position in 96 holes during each microbiotic dilution:

Stoste: the initial soln of medicine after preparation

(once) diluent: 10 times of gained solution of stoste dilution, promptly 100ul stoste and 900ul solvent form

(secondary) redilution liquid: 10 times of gained solution of diluent redilution, promptly 100ul diluent and 900ul solvent form:

Microbiotic name location solution title consumption (ul)

A1 -- --

B1 -- --

No. 17 medicine stostes 180 of C1

No. 17 medicines

No. 17 medicine stostes 90 of D1

Ciprofloxacin

No. 17 medicine stostes 45 of E1

No. 17 medicine diluents 225 of F1

No. 17 medicine diluents 112 of G1

No. 17 medicine diluents 56 of H1

Microbiotic name location solution title consumption (ul)

No. 3 medicine stostes 288 of A2

No. 3 medicine stostes 144 of B2

No. 3 medicine stostes 72 of C2

No. 3 medicines

No. 3 medicine stostes 36 of D2

Amikacin

No. 3 medicine diluents 180 of E2

No. 3 medicine diluents 90 of F2

No. 3 medicine diluents 45 of G2

No. 3 medicine diluents 23 of H2

Microbiotic name location solution title consumption (ul)

No. 16 medicine stostes 480 of A3

No. 16 medicine stostes 240 of B3

No. 16 medicine stostes 120 of C3

No. 16 medicines

No. 16 medicine stostes 60 of D3

Rifampin

No. 16 medicine stostes 30 of E3

No. 16 medicine diluents 150 of F3

No. 16 medicine diluents 75 of G3

No. 16 medicine diluents 38 of H3

Microbiotic name location solution title consumption (ul)

No. 14 medicine stostes 384 of A4

No. 14 medicine stostes 192 of B4

No. 14 medicines

No. 14 medicine stostes 96 of C4

Erythromycin

No. 14 medicine stostes 48 of D4

No. 14 medicine diluents 240 of E4

No. 14 medicine diluents 120 of F4

No. 14 medicine diluents 60 of G4

No. 14 medicine diluents 30 of H4

Microbiotic name location solution title consumption (ul)

No. 12 medicine stostes 384 of A5

No. 12 medicines

No. 12 medicine stostes 192 of B5

Clindamycin

No. 12 medicine stostes 96 of C5

No. 12 medicine stostes 48 of D5

No. 12 medicine diluents 240 of E5

No. 12 medicine diluents 120 of F5

No. 12 medicine diluents 60 of No. 12 medicine G5

No. 12 medicine diluents 30 of clindamycin H5

Microbiotic name location solution title consumption (ul)

No. 11 medicine stostes 684 of A6

No. 11 medicine stostes 342 of B6

No. 11 medicine stostes 171 of C6

No. 11 medicines

No. 11 medicine stostes 86 of D6

SMZ

No. 11 medicine stostes 42 of E6

No. 11 medicine diluents 210 of F6

No. 11 medicine diluents 105 of G6

No. 11 medicine diluents 53 of H6

Microbiotic name location solution title consumption (ul)

No. 15 medicine stostes 360 of A7

No. 15 medicine stostes 180 of B7

No. 15 medicine stostes 90 of C7

No. 15 medicines

No. 15 medicine stostes 45 of D7

Vancomycin

No. 15 medicine diluents 225 of E7

No. 15 medicine diluents 112 of F7

No. 15 medicine diluents 56 of G7

No. 15 medicine diluents 28 of H7

Microbiotic name location solution title consumption (ul)

No. 19 medicine stostes 45 of A8

No. 19 medicine diluents 225 of B8

No. 19 medicine diluents 112 of C8

No. 19 medicines

No. 19 medicine diluents 56 of D8

Penicillin

No. 19 medicine redilution of E8 liquid 280

No. 19 medicine redilution of F8 liquid 140

No. 19 medicine redilution of G8 liquid 70

No. 19 medicine redilution of H8 liquid 35

Microbiotic name location solution title consumption (ul)

No. 13 medicine stostes 240 of A9

No. 13 medicine stostes 120 of B9

No. 13 medicine stostes 60 of C9

No. 13 medicines

No. 13 medicine stostes 30 of D9

Oxazacillin

No. 13 medicine diluents 150 of E9

No. 13 medicine diluents 75 of F9

No. 13 medicine diluents 38 of G9

No. 13 medicine diluents 19 of H9

Microbiotic name location solution title consumption (ul)

No. 6 medicine stostes 288 of A10

No. 6 medicine stostes 144 of B10

No. 6 medicine stostes 72 of C10

No. 6 medicines

No. 6 medicine stostes 36 of D10

Kefzol

No. 6 medicine diluents 180 of E10

No. 6 medicine diluents 90 of F10

No. 6 medicine diluents 45 of G10

No. 6 medicine diluents 23 of H10

Microbiotic name location solution title consumption (ul)

No. 41 medicine stostes 288 of A11

No. 41 medicine stostes 144 of B11

No. 41 medicines

No. 41 medicine stostes 72 of C11

Cefaclor

No. 41 medicine stostes 36 of D11

No. 41 medicine diluents 180 of E11

No. 41 medicine diluents 90 of F11

No. 41 medicine diluents 45 of G11

No. 41 medicine diluents 23 of H11

Microbiotic name location solution title consumption (ul)

No. 10 medicine stostes 288 of A12

No. 10 medicine stostes 144 of B12

No. 10 medicine stostes 72 of C12

No. 10 medicines

No. 10 medicine stostes 36 of D12

Ceftriaxone

No. 10 medicine diluents 180 of E12

No. 10 medicine diluents 90 of F12

No. 10 medicine diluents 45 of G12

No. 10 medicine diluents 23 of H12.

4, want the preparation method of 2 described a kind of staphylococcus drug sensitive battens according to right, the staphylococcus drug sensitive batten that it is characterized in that making by this method is behind the bacterium bacteria suspension that carries out bacterium operation adding 100ul, and the microbiotic ultimate density is as follows in the lath: unit is: ug/ml

MS lath susceptibility concentration gradient table

Annotate: be the scope between medicaments insensitive and Nai Yue between the shade.