CN1114699C - Prepn of animal's fast drug susceptability test paper - Google Patents

Abstract

The present invention relates to a method for preparing fast drug sensitive test paper for animals, which comprises technological steps, such as preparation of sterilizing filter paper sheets, preparation of a drug solution, preparation of drug sensitive test paper, etc. Compared with the prior art, the present invention has the advantages of simple technology and low cost; when the product of the present invention is used for testing, the operation is convenient, and the result is accurate; economic loss caused by blind use, excessive use and wrong use of drugs is reduced, and the damage of drug residues of animal products to human bodies is reduced. The present invention provides the extremely direct basis for reasonable drug use.

Description

The preparation method of animal's fast drug susceptability test paper

The present invention relates to the preparation method of drug susceptability test paper, relate in particular to a kind of preparation method of animal's fast drug susceptability test paper.

At present, the edible livestock and poultry of the domestic overwhelming majority all are to adopt intensification stable breeding and the mode of raising in cages to raise, and epidemic prevention is cured the disease and become sixty-four dollar question in its feeding process.In recent years, because the abuse of antibacterials in livestock industry is cultured, Resistant strain also increases thereupon greatly, as diseases such as chicken colibacillosis, white dysentery, chronic respiratory tract disease, staphylococcosis and cyanomycosis and yellow scour of piglet, dysentery characterized by white mucous stool, because of chemical sproof generation, because of the raiser lacks the correct judgement that effective antibacterials are selected, in use blindly the administration phenomenon is quite general simultaneously.The main consequence that this phenomenon causes is: 1. the livestock-raising family has increased unnecessary financial loss; 2. increase the resistance of livestock and poultry, the control cost is risen; 3. antibacterials exceed standard residual seriously in the livestock and poultry animal body, and there is suitable detrimental effect the edible back of people to human body.In order to address the above problem, some colleges and universities and scientific research institution under lab adopt research methods such as steel ring method to carry out drug sensitive test, but that these methods prepare is loaded down with trivial details, and the working conditions harshness can't solve the practical problems that numerous raisers face.

Purpose of the present invention is exactly to provide a kind of for the defective that overcomes above-mentioned prior art existence can judge quickly and accurately that pathogenic bacterium are to the preparation method of the animal's fast drug susceptability test paper of antibiotic drug susceptibility in the animal body.

Purpose of the present invention can be achieved through the following technical solutions: the preparation method of animal's fast drug susceptability test paper, be characterized in, comprise the technological process of preparation of preparation, the drug susceptability test paper of preparation, the drug solution of the filter paper of sterilizing, its concrete processing step is as follows:

The first step, the preparation of sterilization filter paper:

A. get the prototype qualitative filter paper, it is processed into the two-sided filter paper that has word marking of diameter 4～8mm, the weighing branch installs in the cillin bottle, 100～150 every bottle;

B. with the filter paper after the packing 100～140 ℃ of following dry sterilizations 1～2 hour;

Second step: the preparation of drug solution:

To the susceptibility of antibacterials and the drug level in the blood, antibacterials are made into certain density solution through weighing, dissolving, constant volume, dilution according to pathogenic bacterium.

The 3rd step, the preparation of drug susceptability test paper:

A. with suction pipe the soup that configures is added on the sterilized filter paper, per 100 add 0.5～0.8 milliliter of soup, soak into the back fully and take out, and dry 20～24 hours down at 37～40 ℃;

B. the drug susceptability test paper that will dry in gnotobasis is inserted in the medicine bottle, with mouthful sealing of sealing with wax;

C. medicine bottle is labelled and carry out outer packaging, preserve standby down for 4～10 ℃.

Described antibacterials are selected from a kind of in the following medicine, and these medicines comprise:

A. penicillin sodium salt: 1000～2000iu/ml;

B. sodium ampicillin: 2～4mg/ml;

C. Vetstrep: 200～300iu/ml;

D. sulphuric acid kanamycin: 0.2～0.4mg/ml;

E. gentamicin sulphate: 100～300iu/ml;

F. amikacin sulfate: 0.05～0.1mg/ml;

G. neomycinsulphate: 0.06～0.15mg/ml;

H. erythromycin lactobionate: 0.2～0.4mg/ml;

I. paraxin: 0.3～0.6mg/ml;

J. sharp proceomycin: 0.4～0.8mg/ml;

K. Vibramycin hydrochloride: 1～2mg/ml;

L. ciprofloxacin HCl: 0.3～0.6mg/ml;

M. star is killed in the promise of hydrochloric acid grace: 0.3～0.6mg/ml;

N. nicotinic acid promise fluorine kills star: 0.3～0.6mg/ml;

O. U 10149a: 0.3～0.6mg/ml;

P. sulfamido: 4～8mg/ml;

G. Ofloxacine USP 23: 0.3～0.5mg/ml;

R. Zinc-bacitracin: 0.1～0.3mg/ml;

S. colistine sulfate: 0.2～0.4mg/ml;

T. fumaric acid tiamulin: 0.05～0.1mg/ml.

By implementing technique scheme, make the present invention obtain the drug susceptability test paper of pathogenic bacterium in a kind of object that can judge quickly and accurately, it is simplified to quick, the easy testing method that general raiser can both grasp to drug sensitive test process that just can carry out, complicated under quite technology and material conditions only.The present invention is simple to operate, easy to use, the result is accurate, expense is cheap, be applicable to that pathogenic bacterium are to the mensuration of antibiotic drug susceptibility in the disease of various zoogenetic infections on the breeding production, use product of the present invention to test, reduce blindness, excess, abused the loss that medicine brings, reduced the harm of animal products drug residue, for rational use of drug provides the most direct foundation.

Below by specific embodiment, the present invention is described in further detail.

Embodiment 1

Present embodiment is the preparation methods of the interior pathogenic bacterium of a kind of animal body to the salt-sensitive drug susceptability test paper of benzylpenicillin sodium.It is divided into following processing step:

The first step, the prototype qualitative filter paper is at first got in the preparation of filter paper, the diameter that adopts the spray printing technology that it is processed into the mark of two-sided having " penicillin sodium salt " is the filter paper sequin of 6mm, the weighing branch installs in the cillin bottle, and 100 every bottle, then 120 ℃ of following dry sterilizations 2 hours; In second step, the preparation of drug solution takes by weighing the standard substance 8 * 10 of penicillin sodium salt ⁴IU adds the small amounts of phosphoric acid damping fluid with after the medicine dissolving, inserts constant volume in the 50ml volumetric bottle, then it is diluted to 1600IU/ml; The 3rd step, the preparation of drug susceptability test paper, at first with suction pipe the benzylpenicillin sodium salt spices liquid that configures is added on the above-mentioned sterilized filter paper, per 100 add soup 0.6ml, soak into the back fully and take out, dried 24 hours down at 38 ℃, the drug susceptability test paper that to dry in sterilized environment is inserted in the medicine bottle then, with mouthful sealing of sealing with wax, carries out outer packaging at last after sticking the printed words that indicate penicillin sodium salt on the medicine bottle, insert refrigerator, preserve standby down at 4～10 ℃.

The drug susceptability test paper that the present invention makes generally will use with vessel together in conjunction with measuring with substratum, mensuration, and is easy to use for making the raiser, also drug susceptability test paper, substratum, mensuration vessel can be contained in the packing box.Introduce their preparation method below respectively: (1) measures the preparation with substratum, substratum is divided into ordinary nutrient agar substratum, serum nutrient agar and potato agar substratum, substratum generally is to place the substratum ware, the preparation of substratum can be divided into following a few step again, at first buy standard nutrient agar medium product, dissolve then, heat, in sterile environment, evenly place substratum ware cooling and shaping again, if any particular requirement, can add additional substances, as serum etc., seal behind the sterilization packaging bag of packing at last, insert packing box.(2) measure the preparation of using vessel, mensuration comprises glass rod, suction pipe, bottle stopper etc. with vessel, requirement is chosen in medical Glassware Factory, rubber item factory and the plastics factory of medical administrative authority fixed point, these vessel are sterilized after cleaning, drying, and in sterilized environment combination packaging, insert packing box.(3) drug susceptability test paper is measured the preparation of packing box, gets one and has certain volumetrical box body, and the drug susceptability test paper of packing into successively, substratum ware, test vessel and working instructions etc. are promptly finished after the outer box sealing.

For the present invention more clearly is described, the using method of the drug susceptability test paper box that brief description once makes according to the present invention.

1. taking-up medium cassette is got wherein bag culture medium ware opening;

2. the inoculation pathogenic bacterium are inoculated into suspicious pathogenic bacterium on the ready-made plate substratum;

3. the placement of drug susceptability test paper behind the inoculated bacteria, is taken out drug susceptability test paper and is placed on the ware from bottle, every ware can evenly be placed 7 of drug susceptability test papers at most;

4. cultivate, the plate substratum of placing drug susceptability test paper was cultivated 24 hours in 37 ℃ thermostat container;

5. observe and judge criterion:

Antibacterial circle diameter (mm)	Criterion
		≥15 11～15mm ≤10mm	Extremely sensitive medium sensitivity resistance

Various pathogenic bacterium are to the susceptibility difference of different antibacterials, and the different strains of same pathogenic bacterium is also variant to the susceptibility of different antibacterials.Separate pathogenic bacterium after cultivating, use the drug susceptability test paper of different pharmaceutical that flora is carried out the sensitivity test, can obviously observed antibacterial phenomenon with naked eyes, make the judgement that kill and wound susceptibility of flora to different antibacterials, the result of drug susceptability test paper mensuration can provide a fast and reasonable dosage regimen in view of the above, improve curative effect, reduce drug cost, for preventing and treat the approach that infectious diseases has found a reliable science in the aquaculture.

Embodiment 2

Present embodiment is the preparation methods of the interior pathogenic bacterium of animal body to the drug susceptability test paper of sodium ampicillin sensitivity.It comprises following processing step: get at first that to be processed into diameter be that the filter paper of 4mm was 100 ℃ of following dry sterilizations 2 hours, the sodium ampicillin of getting concentration then and be 2mg/ml is added on the sterilized filter paper, per 100 add 0.5ml, soak into the back fully and take out, and dry 20 hours down at 37 ℃; The last drug susceptability test paper that in sterilized environment, will dry insert seal with wax in the medicine bottle standby.All the other are identical with embodiment 1 about technology.

Embodiment 3

Present embodiment is the preparation methods of the interior pathogenic bacterium of animal body to the drug susceptability test paper of Vetstrep sensitivity.It comprises following processing step: get at first that to be processed into diameter be that the filter paper of 8mm was 140 ℃ of following dry sterilizations 1 hour, the Vetstrep of getting concentration then and be 250iu/ml is added on the sterilized filter paper, per 100 add 0.8ml, soak into the back fully and take out, and dry 24 hours down at 40 ℃; The last drug susceptability test paper that in sterilized environment, will dry insert seal with wax in the medicine bottle standby.All the other are identical with embodiment 1 about technology.

Embodiment 4～20

Following examples be in the animal body pathogenic bacterium respectively to the preparation method of the drug susceptability test paper of 17 kinds of antibacterials sensitivities such as sulphuric acid kanamycin.

The antibacterials and the concentration thereof of its use are as shown in the table:

Sulphuric acid kanamycin	0.2～0.4mg/ml
		Gentamicin sulphate	100～300 iu/ml
Amikacin sulfate	0.05～0.1mg/ml
		Neomycinsulphate	0.06～0.15mg/ml
Erythromycin lactobionate	0.2～0.4mg/ml
		Paraxin	0.3～0.6mg/ml
Sharp proceomycin	0.4～0.8mg/ml
		Vibramycin hydrochloride	1～2mg/ml
Ciprofloxacin HCl	0.3～0.6mg/ml
		Star is killed in the promise of hydrochloric acid grace	0.3～0.6mg/ml
Nicotinic acid promise fluorine kills star	0.3～0.6mg/ml
		U 10149a	0.3～0.6mg/ml
Sulfamido	4～8mg/ml
		Ofloxacine USP 23	0.3～0.5mg/ml
Zinc-bacitracin	0.1～0.3mg/ml
		Colistine sulfate	0.2～0.4mg/ml
Fumaric acid tiamulin	0.05～0.1mg/ml

Above-mentioned antibacterials are added on the sterilized filter paper, and all the other technologies are identical with embodiment 1.

Claims (2)

1, the preparation method of animal's fast drug susceptability test paper is characterized in that, comprises the technological process of preparation of preparation, the drug susceptability test paper of preparation, the drug solution of the filter paper of sterilizing, and its concrete processing step is as follows:

The first step, the preparation of sterilization filter paper:

A. get the prototype qualitative filter paper, it is processed into the two-sided filter paper that has word marking of diameter 4～8mm, the weighing branch installs in the cillin bottle, 100～150 every bottle;

B. with the filter paper after the packing 100～140 ℃ of following dry sterilizations 1～2 hour;

Second step: the preparation of drug solution:

To the susceptibility of antibacterials and the drug level in the blood, antibacterials are made into certain density solution through weighing, dissolving, constant volume, dilution according to pathogenic bacterium.

The 3rd step, the preparation of drug susceptability test paper:

A. with suction pipe the soup that configures is added on the sterilized filter paper, per 100 add 0.5～0.8 milliliter of soup, soak into the back fully and take out, and dry 20～24 hours down at 37～40 ℃;

B. the drug susceptability test paper that will dry in gnotobasis is inserted in the medicine bottle, with mouthful sealing of sealing with wax;

C. medicine bottle is labelled and carry out outer packaging, preserve standby down for 4～10 ℃.

2, the preparation method of animal's fast drug susceptability test paper according to claim 1 is characterized in that, described antibacterials are selected from a kind of in the following medicine, and these medicines comprise:

A. penicillin sodium salt: 1000～2000iu/ml;

B. sodium ampicillin: 2～4mg/ml;

C. Vetstrep: 200～300iu/ml;

D. sulphuric acid kanamycin: 0.2～0.4mg/ml;

E. gentamicin sulphate: 100～300iu/ml;

F. amikacin sulfate: 0.05～0.1mg/ml;

G. neomycinsulphate: 0.06～0.15mg/m1;

H. erythromycin lactobionate: 0.2～0.4mg/ml;

I. paraxin: 0.3～0.6mg/ml;

J. sharp proceomycin: 0.4～0.8mg/ml;

K. Vibramycin hydrochloride: 1～2mg/ml;

L. ciprofloxacin HCl: 0.3～0.6mg/ml;

M. star is killed in the promise of hydrochloric acid grace: 0.3～0.6mg/ml;

N. nicotinic acid promise fluorine kills star: 0.3～0.6mg/ml;

O. U 10149a: 0.3～0.6mg/ml;

P. sulfamido: 4～8mg/ml;

G. Ofloxacine USP 23: 0.3～0.5mg/ml;

R. Zinc-bacitracin: 0.1～0.3mg/ml;

S. colistine sulfate: 0.2～0.4mg/ml;

T. fumaric acid tiamulin: 0.05～0.1mg/ml.