CN110016494A - A kind of haemophilus drug sensitivity testing in vitro detector bar - Google Patents
A kind of haemophilus drug sensitivity testing in vitro detector bar Download PDFInfo
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- CN110016494A CN110016494A CN201910332051.7A CN201910332051A CN110016494A CN 110016494 A CN110016494 A CN 110016494A CN 201910332051 A CN201910332051 A CN 201910332051A CN 110016494 A CN110016494 A CN 110016494A
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- 230000035945 sensitivity Effects 0.000 title claims abstract description 43
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- 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
- C12Q1/18—Testing for antimicrobial activity of a material
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Abstract
The invention discloses a kind of haemophilus drug sensitivity testing in vitro detector bars, it is strip, growth factor needed for being evenly provided with severe bacteria growth in detection zone on one side, and compartment of terrain is successively arranged the antibacterials of multiple proportions decrement in order along its length, the another side of detector bar is respectively equipped with scale and minimum inhibitory concentration value corresponding with antimicrobial object amount in corresponding position；Antibacterials on detector bar are after carrying out doubling dilution to detection drug, the drug solution of resulting various concentration to be coated on the corresponding position of detector bar, re-dry forms.Severe bacteria includes haemophilus, and growth factor includes the V factor and the X factor needed for haemophilus growth.The width of detector bar is preferably 3mm.Haemophilus drug sensitivity testing in vitro detector bar provided by the invention can detect severe bacteria drug sensitivity testing in vitro in the M-H agar plate of the most basic indispensability of Microbiological Lab, and severe bacteria susceptibility problem cannot be detected on conventional applied culture base by, which making, is resolved.
Description
Technical field
The present invention relates to a kind of drug sensitive test tools, and in particular, to a kind of haemophilus drug sensitivity testing in vitro detector bar.
Background technique
In-vitro antibacterial medicament sensitivity test abbreviation drug sensitive test (AST), that is, resistance test, refer to and measure in vitro
Drug is antibacterial or the test of sterilizing ability.It is intended to understand pathogenic microorganism to the sensitivities of various antibiotic (or tolerance) degree, with
Clinical rational is instructed to select the microbiology test of antibiotic medicine.
Various pathogenic bacteria are different to the sensibility of different antibacterials, and the different strains of same bacterium are to different antimicrobials
The sensibility of object is also variant.For a long time, the generation of various pathogenic bacteria drug resistances loses various Common Antibiotics often
Drug effect, and drug cannot be grasped well to the susceptibility of bacterium, so a correct drug sensitive test is as a result, for clinic
Doctor selects the reference of antibacterials, and improves curative effect.If a kind of antibiotic can be inhibited with the dosage of very little, kill and cause a disease
Bacterium then claims this kind of pathogenic bacteria to the antibiotic " sensitivity ".Conversely, being then known as " insensitive " or " drug resistance ".To understand pathogenic bacteria pair
Which kind of antibiotic sensitivity is reduced blindness, should often be carried out drug sensitive test with the rational use of medicines.Substantially method is for it: from patient
Infection site takes the sample containing pathogenic bacteria, is seeded on culture medium appropriate, cultivates under certain condition；It will be stained with respectively simultaneously
There are the scraps of paper of a certain amount of various antibiotic to be attached to media surface (or with stainless steel coil, interior amount antibiotic solution blow-down), culture
Result is observed after a certain period of time.Since sensitivity of the pathogenic bacteria to various antibiotic is different, just occur around drug paper disk
Different size of inhibition pathogen growth and form " sky circle ", referred to as inhibition zone.Inhibition zone size and pathogenic bacteria are to various antibiosis
The sensitivity of element is proportional.Then antibiotic can targetedly be selected according to test result.According to American National
The standard that the Clinical Laboratory Standard committee (NCCLS) is recommended in the recent period, to non-severe oxygen bacterium, (enterobacteriaceae lactobacteriaceae, verdigris are false single
Born of the same parents bacterium and other non-enteric section bacillus, staphylococcus bacteria, Enterococcus) and severe oxygen bacterium (hemophilus bacterium, leaching
Sick Neisseria, streptococcus pneumonia and other streptococcus) select the choice drug (A group antibiotic) of conventional drug sensitive test or clinic to make
Major antibiotics (B group antibiotic) carry out drug sensitive test.
Abuse of antibiotics at present causes anti-medicine bacterium to increase, or even largely uses broad-spectrum antibiotic because long-term, in contact element just
Normal microorganism, loses the mutual restrictive function of microorganism, to keep non-pathogenic bacteria some rare or under normal circumstances a large amount of
The case where breeding, causing so-called " superinfection " is occurred repeatedly, and causes artificial difficulty to treatment.Therefore, it advocates and uses susceptibility
Test, adheres to that the rational use of medicines is particularly significant.
There are mainly three types of the modes of bacterium drug sensitivity testing in vitro: agar diffusion method, agar dilution and E-test method.
The first is known as agar diffusion method.Specific method is the bacterium that respective concentration is made in the bacterium to be detected that will be isolated
Liquid after bacterium solution is applied on the agar plate of adaptation, is pasted containing the round paper slightly larger than 6mm for being detected antibacterials
Piece will appear the non-growing circle of bacterium after proper temperature, gas, time culture around the scraps of paper, this circle size can
Reflect bacterium to the sensibility of drug.The method derive from agar dilution, it is easy to operate with its, detection flexibly etc. made extensively
With.The disadvantage is that first, inhibition zone size and its sensitivity relationship that individual bacteria occur in individual drugs are inconsistent.Second, this
For two stage approach, it can only accomplish qualitative detection.Third, there are no sensitivity metrics for many bacteriums.Fourth, although being clinically separated
Most bacteriums can detect on MH agar plate, and there are also some bacteriums can only respectively special culture media detects at its.Bloodthirsty bar
Bacterium is one of them.It is encountered since haemophilus is less, and the culture medium shelf-life needed for its susceptibility is very short, so, susceptibility inspection
Survey is just very chaotic, some, which have no alternative but way result accuracy, cannot be guaranteed.
Second be known as agar dilution.The method is the methodical basis of institute.Specific method is that antibacterials are added
Agar plate is made in culture medium suitable for being detected bacterial growth, a drug need to generally do 15 various concentration plates, drug
Concentration presses doubling dilution, and such as first 512 μ g/ml, second 256 μ g/ml, the 3rd 128 μ g/ml are arranged successively.It will be detected
Bacterial solution is distinguished in dibbling to each plate, and after the culture of proper temperature, gas and time, bacterium cannot inoculate a life
Long plate contained drug concentration is minimum inhibitory concentration MIC (minimal inhibit concentration).The method
It is to have the characteristics that quantitative and once can detect multiple bacteriums a little.The disadvantage is that most drugs are unstable in the medium and the shelf-life
Shorter, operation is inconvenient, so clinical labororatory does not use this method for routine testing substantially.The method derives
Broth dilution method and micro broth dilution method.The method is calibrated for retrospective scientific research and other methods.Same the method is not yet
It is usually used in haemophilus detection suitable day.
The third is E-test method.The method belongs to newer method, and with the immediate method of the application.Specific method
It is containing an antibacterials on bar shaped carrier, medication amount successively increases and decreases on item, the quite minimum suppression of identification of medications on item
Such as sequence 512,256,128,64 until 0.03 μ g/ml, that is, 15 MIC doubling dilution values, this is E-test to bacteria concentration
Item.After bacterium solution is applied on the agar plate of adaptation, Eest item is pasted, after proper temperature, gas, time culture,
It will appear the non-growing oval circle of bacterium around item, the corresponding number of oval circle point is the MIC of this bacterium.The method has
First two advantage.Disadvantage different bacterium needs respective special culture media.The haemophilus special culture media shelf-life is very short, actually makes
With being equally inconvenient.
Summary of the invention
The object of the present invention is to provide a kind of drug sensitivity testing in vitro detector bar, growth factor needed for joined severe bacteria makes
Severe bacteria susceptibility problem cannot be detected on conventional applied culture base to be resolved.
In order to achieve the above object, the present invention provides a kind of haemophilus drug sensitivity testing in vitro detector bars, wherein described
Detector bar is strip, and one side is evenly provided with the required growth factor of severe bacteria growth in detection zone, and along length side
The antibacterials that multiple proportions is reduced in order are successively arranged to compartment of terrain, the another side of detector bar is respectively equipped with scale in corresponding position
And minimum inhibitory concentration (MIC) value corresponding with antimicrobial object amount；Antibacterials on the detector bar are to detection medicine
Object carry out doubling dilution after, the drug solution of resulting various concentration is coated on the corresponding position of detector bar, re-dry and
At.
Above-mentioned haemophilus drug sensitivity testing in vitro detector bar, wherein the severe bacteria includes haemophilus, described
Growth factor includes the V factor and the X factor needed for haemophilus growth.Contain suitable severe bacteria for be suitble to the growth of severe bacteria
The solution of growth factor is uniformly coated on the one side of detector bar, then is dried；The mass concentration of the V factor and the X factor in solution
Preferably 0.03%~0.05%, it is formulated using distilled water.The V factor (V facto) is hemophilus
(Haemophilus) factor necessary to certain kinds (such as Hemophilus influenzae H.influenza) are grown.The V factor is Coenzyme I
(i.e. nicotinamide adenine dinucleotide, NAD) or Coenzyme I I (nicotinamide-adenine dinucleotide phosphate, NADP).The X factor (X
Factor) it is a kind of chemokines, is necessary to certain kinds of bacterial growths of Haemophilus spp, it can be by hematin
(Hemin) it replaces.Haemophilus needs the presence of the X factor and the V factor that can just grow, common meat soup and the training of common agar
Feeding base cannot be grown.
Above-mentioned haemophilus drug sensitivity testing in vitro detector bar, wherein the width of the detector bar is 3mm, than foreign countries
The 4mm of import is more suitable for.
Above-mentioned haemophilus drug sensitivity testing in vitro detector bar, wherein compartment of terrain along its length on the detector bar
Solution concentration gradient equipped with 8~15 doubling dilutions.Detector bar length is related with detection range, preferably 8~10 gradients
Clinical labororatory's requirement can be fully achieved, 15 gradients or more can also be done, but detector bar can mistake after more than 15 gradients
Long, agar plate needed for detecting must be special, only applies in scientific research.
Above-mentioned haemophilus drug sensitivity testing in vitro detector bar, wherein the length of each concentration gradient on the detector bar
Degree is 0.25mm, and adjacent concentration gradient is preferably spaced more than or equal to 0.25mm.
Above-mentioned haemophilus drug sensitivity testing in vitro detector bar, wherein the solution concentration gradient of the doubling dilution exists
The minimum inhibitory concentration that the another side of detector bar is correspondingly arranged successively is preferably 512 μ g/ml, 256 μ g/ml, 128 μ g/ml, 64 μ
G/ml is until 0.03 μ g/ml, 15 MIC doubling dilution values altogether.
Above-mentioned haemophilus drug sensitivity testing in vitro detector bar a, wherein face paste of solution is coated on the detector bar
On the Agar Plating (MHA) for being coated with detected severe bacteria, the one side equipped with scale and minimum inhibitory concentration value
Upward.
Above-mentioned haemophilus drug sensitivity testing in vitro detector bar, wherein the severe bacteria on the Agar Plating
After proper temperature, gas, time conditions are cultivated, occurs a non-growing oval of bacterium around detector bar
Circle, the corresponding scale of oval circle point and minimum inhibitory concentration value are the minimum inhibitory concentration of the bacterium.Due to growth factor
From detector bar, if being free of antibacterials on detector bar, it is better apart from the nearlyr growth of detector bar that detection bacterium growth will appear, and leads
Cause seems and incomplete oval, but the point differentiated is detector bar closest approach, has no effect on final judgement.Preferably, it sticks
It is cultivated 15~18 hours after detector bar, then reads Mlc.
Haemophilus drug sensitivity testing in vitro detector bar provided by the invention has the advantage that
The present invention can detect severe bacteria drug sensitivity testing in vitro in the MH agar plate of the most basic indispensability of Microbiological Lab.
It solves clinical labororatory's embarrassing phenomenon when encountering severe bacteria, ensure that inspection result authenticity, overcome severe bacteria detection
The deficiency of aspect.
Detailed description of the invention
Fig. 1 is the schematic diagram of haemophilus drug sensitivity testing in vitro detector bar of the invention.
Fig. 2 uses schematic diagram for haemophilus drug sensitivity testing in vitro detector bar of the invention.
Fig. 3 is the interpretation schematic diagram of haemophilus drug sensitivity testing in vitro detector bar of the invention.
Fig. 4 is the reading schematic diagram of haemophilus drug sensitivity testing in vitro detector bar of the invention.
Specific embodiment
A specific embodiment of the invention is further described below in conjunction with attached drawing.
As shown in Figure 1, haemophilus drug sensitivity testing in vitro detector bar provided by the invention, which is strip,
Growth factor needed for being evenly provided with the growth of severe bacteria in detection zone on one side, and along its length compartment of terrain be successively arranged by
The antibacterials of sequence multiple proportions decrement, the another side of detector bar corresponding position be respectively equipped with scale and with antimicrobial object amount pair
The minimum inhibitory concentration value answered；Antibacterials on detector bar are after carrying out doubling dilution to detection drug, by resulting difference
The drug solution of concentration is coated on the corresponding position of detector bar, and re-dry forms.
Severe bacteria includes haemophilus, and growth factor includes the V factor and the X factor needed for haemophilus growth.
The width of detector bar is preferably 3mm.
Compartment of terrain is equipped with the solution concentration gradient of 8~15 doubling dilutions along its length on detector bar.It is every on detector bar
The length of a concentration gradient is 0.25mm.
The solution concentration gradient of doubling dilution is followed successively by 512 in the minimum inhibitory concentration that the another side of detector bar is correspondingly arranged
μ g/ml, 256 μ g/ml, 128 μ g/ml, 64 μ g/ml are until 0.03 μ g/ml.
It is coated with being attached on the Agar Plating for being coated with detected severe bacteria on one side for solution on detector bar, is equipped with
The one side of scale and minimum inhibitory concentration value is upward.Severe bacteria on Agar Plating is after culture, in detector bar week
The existing non-growing oval circle of a bacterium is crossed, the corresponding scale of oval circle point and minimum inhibitory concentration value are the bacterium
Minimum inhibitory concentration.
Haemophilus drug sensitivity testing in vitro detector bar provided by the invention is further described below with reference to embodiment.
Embodiment 1
A kind of haemophilus drug sensitivity testing in vitro detector bar, the detector bar are strip, and one side is uniform in detection zone
Growth factor needed for ground is equipped with the growth of severe bacteria, and compartment of terrain is successively arranged the antibacterial of multiple proportions decrement in order along its length
The another side of drug, detector bar is respectively equipped with scale and minimum inhibitory concentration corresponding with antimicrobial object amount in corresponding position
(MIC) value；Antibacterials on detector bar be to detection drug carry out doubling dilution after, the drug of resulting various concentration is molten
Liquid is coated on the corresponding position of detector bar, and re-dry forms.
Severe bacteria includes haemophilus, and growth factor includes the V factor and the X factor needed for haemophilus growth.It will be suitble to
The solution containing suitable severe bacteria growth factor of severe bacteria growth is uniformly coated on the one side of detector bar, then is dried；
The mass concentration of the V factor and the X factor is preferably 0.03%~0.05% in solution, is formulated using distilled water.The V factor (V
Facto) be hemophilus (Haemophilus) certain kinds (such as Hemophilus influenzae H.influenza) growths institute it is required
The factor.The V factor is Coenzyme I (i.e. nicotinamide adenine dinucleotide, NAD) or Coenzyme I I (nicotinamide adenine dinucleotide
Phosphoric acid, NADP).The X factor (X factor) is a kind of chemokines, is necessary to certain kinds of bacterial growths of Haemophilus spp,
It can be replaced by hematin (Hemin).Haemophilus needs the presence of the X factor and the V factor that can just grow, common meat soup
It cannot be grown with common agar medium.
The width of detector bar is 3mm, and the 4mm than external import is more suitable for.
Compartment of terrain is equipped with the solution concentration gradient of 8~15 doubling dilutions along its length on detector bar.Detector bar length
It is related with detection range, preferably 8~10 gradients can be fully achieved clinical labororatory requirement, can also do 15 gradients or
More, but after more than 15 gradients detector bar can be too long, and agar plate needed for detecting must be special, only applies in scientific research.
The length of each concentration gradient is 0.25mm on detector bar, and being preferably spaced for adjacent concentration gradient is greater than or waits
In 0.25mm.
The minimum inhibitory concentration that the solution concentration gradient of doubling dilution is correspondingly arranged in the another side of detector bar is successively preferred
It is 512 μ g/ml, 256 μ g/ml, 128 μ g/ml, 64 μ g/ml until 0.03 μ g/ml, 15 MIC doubling dilution values altogether.
Solution is coated on detector bar is attached to the Agar Plating (MHA) for being coated with detected severe bacteria on one side
On, the one side equipped with scale and minimum inhibitory concentration value is upward.
Severe bacteria on Agar Plating is after proper temperature, gas, time conditions are cultivated, in detector bar
There is the non-growing oval circle of a bacterium in surrounding, and the corresponding scale of oval circle point and minimum inhibitory concentration value are that this is thin
The minimum inhibitory concentration of bacterium.Since growth factor comes from detector bar, if being free of antibacterials on detector bar, detection bacterium grows meeting
Appearance distance detector bar is closer, and growth is better, causes to seem and incomplete oval, but the point differentiated is detector bar closest approach,
Have no effect on final judgement.Preferably, it is cultivated 15~18 hours after sticking detector bar, then reads Mlc.Referring to fig. 2~
Shown in 4.
Haemophilus drug sensitivity testing in vitro detector bar provided by the invention joined severe bacteria institute on the basis of E-test item
Growth factor is needed, severe bacteria drug sensitivity testing in vitro can be detected in the MH agar plate of the most basic indispensability of Microbiological Lab, made
Severe bacteria susceptibility problem cannot be detected on conventional applied culture base to be resolved.
It is discussed in detail although the contents of the present invention have passed through above preferred embodiment, but it should be appreciated that above-mentioned
Description is not considered as limitation of the present invention.After those skilled in the art have read above content, for of the invention
A variety of modifications and substitutions all will be apparent.Therefore, protection scope of the present invention should be limited to the appended claims.
Claims (8)
1. a kind of haemophilus drug sensitivity testing in vitro detector bar, which is characterized in that the detector bar is strip, and one side is being examined
Growth factor needed for region is evenly provided with severe bacteria growth is surveyed, and compartment of terrain is successively arranged multiple proportions in order along its length
The antibacterials of decrement, the another side of detector bar are respectively equipped with scale and corresponding with antimicrobial object amount minimum in corresponding position
Inhibitory concentration values；
Antibacterials on the detector bar are after carrying out doubling dilution to detection drug, by the drug of resulting various concentration
On the corresponding position of detector bar, re-dry forms solution coating.
2. haemophilus drug sensitivity testing in vitro detector bar as described in claim 1, which is characterized in that the severe bacteria includes
Haemophilus, the growth factor include the V factor and the X factor needed for haemophilus growth.
3. haemophilus drug sensitivity testing in vitro detector bar as described in claim 1, which is characterized in that the width of the detector bar
Degree is 3mm.
4. haemophilus drug sensitivity testing in vitro detector bar as claimed in claim 3, which is characterized in that edge on the detector bar
Length direction compartment of terrain is equipped with the solution concentration gradient of 8~15 doubling dilutions.
5. haemophilus drug sensitivity testing in vitro detector bar as claimed in claim 4, which is characterized in that every on the detector bar
The length of a concentration gradient is 0.25mm.
6. haemophilus drug sensitivity testing in vitro detector bar as claimed in claim 4, which is characterized in that the doubling dilution
Solution concentration gradient is followed successively by 512 μ g/ml, 256 μ g/ml, 128 in the minimum inhibitory concentration that the another side of detector bar is correspondingly arranged
μ g/ml, 64 μ g/ml are until 0.03 μ g/ml.
7. the haemophilus drug sensitivity testing in vitro detector bar as described in any one of claim 1~6, which is characterized in that institute
It is coated with being attached on the Agar Plating for being coated with detected severe bacteria on one side for solution on the detector bar stated, is equipped with scale
Upward with the one side of minimum inhibitory concentration value.
8. haemophilus drug sensitivity testing in vitro detector bar as claimed in claim 7, which is characterized in that the agar plate training
The severe bacteria on base is supported after culture, occurs the non-growing oval circle of a bacterium, oval circle point around detector bar
Corresponding scale and minimum inhibitory concentration value are the minimum inhibitory concentration of the bacterium.
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| CN110904189A (en) * | 2019-12-05 | 2020-03-24 | 湖南长沙天地人生物科技有限公司 | Haemophilus identification drug sensitivity test card and preparation method thereof |
| CN115341013A (en) * | 2022-07-06 | 2022-11-15 | 清华珠三角研究院 | Microbial drug sensitivity detection method, device, equipment and storage medium |
| WO2024110938A1 (en) * | 2022-11-25 | 2024-05-30 | Biomaxima S.A. | Minimal inhibitory concentration (mic) test strip |
| WO2024110939A1 (en) * | 2022-11-25 | 2024-05-30 | Biomaxima S.A. | Minimal inhibitory concentration (mic) test strip |
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