CN104614493A - Natural allergen detection agent preparation instrument and application thereof - Google Patents
Natural allergen detection agent preparation instrument and application thereof Download PDFInfo
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Abstract
The invention provides a natural allergen detection agent preparation instrument and application thereof. The preparation instrument comprises a sample adding groove, a solvent groove, a low-temperature mixing and crushing tank and a low-temperature centrifugal extraction tank; the top of the sample adding groove is provided with a sample adding port, the bottom of the sample adding groove is provided with a sample outlet, the top of the solvent groove is provided with a solvent adding port, and the bottom of the solvent groove is provided with a solvent outlet; the sample outlet of the sample adding groove and the solvent outlet of the solvent groove are respectively communicated with the interior of the low-temperature mixing and crushing tank, the low-temperature mixing and crushing tank is communicated with the low-temperature centrifugal extraction tank through a connecting pipe, and one path of the liquid separated from the connecting pipe is used as a liquid separating outlet for homogenate in the low-temperature mixing and crushing tank; the top of the low-temperature centrifugal extraction tank is also provided with a supernatant outlet, and the bottom is provided with a sediment outlet. The preparation instrument is used for preparing the allergen immersion liquid reagent for clinical allergen internal detection, and has the advantages of short time, low price, no limitation of allergen types and the like.
Description
Technical field
The invention belongs to Allergic skin test device field, particularly relate to a kind of native allergens detection agent preparing instrument and application thereof.
Background technology
Along with the change of environment for human survival and life style, the incidence of disease of anaphylactia raises year by year.Point out in the global white paper (WAO-WHITEON ALLERGY) that the World Health Organization (WHO) (WHO) issues when the new century arrives: anaphylactia becomes global health problem, the global population of about 20% suffers from anaphylactia at present, and it has been known as the worldwide chronic disease of ranking the 6th after disease of cardiovascular system, respiratory disease, diabetes, tumour and psychotic mental illness.Showing great attention to and paying attention to of international community and medical science all circles has been subject to as common disease, frequently-occurring disease.
Anaphylactia belongs to hypersensitivity disease category, and hypersensitivity refers to that body accepts specific antigen and continues to stimulate or same antigen stimulates the reaction of the pathologic immune such as caused dysfunction and (or) tissue damage again.Hypersensitivity is divided into again four types, wherein the hypersensitivity of I type is also called immediate hypersensitivity, that (allergic effect) is former is combined with the IgE of mast cell, basophil cellular surface because of irritated, cause cell degranulation, delivery of biologically active medium, act on effector organ, cause telangiectasis, permeability increase, smooth muscle contraction, glandular secretion enhancing etc., the allergic inflammation reaction caused.The hypersensitivity of IV type is also called delayed allergy, because again meeting with corresponding antigens by the T lymphocyte of sensitization after irritated (allergic effect) primary stimuli, direct killing target cell or generation cytokine profiles, (hypersensitivity of II type is cytotoxicity hypersensitivity, as ABO transfusion reaction to cause the inflammatory reaction based on mononuclear cells infiltration; Type III hypersensitivity is immune complex type hypersensitivity, as serum sickness).
I type and the hypersensitivity of IV type is diseases related is also referred to as anaphylactia, mainly comprises the diseases such as allergic rhinitis, allergic conjunctivitis, allergia nose-nasosinusitis, bronchial astehma, atopic dermatitis, contact dermatitis, severe allergic reaction, food hypersenstivity, nettle rash, angioedema, medicine and biopreparate are irritated, insect toxins is irritated, occupational is irritated, motion relevance is irritated.The generation of above anaphylactia is closely bound up with individual gene form (neurological susceptibility), living environment (anaphylactogen) and life style (risk factor) with development.
At present the research that the mankind cross sensitive gene is still in the junior stage, cannot cut off expression and the modification of gene from essence, and the risk factor in life is progressively familiar with, the measure also gradual perfection of corresponding antagonism risk factor, as haze weather wears mouth mask, shrimp crab allergy forbids eating.But the investigation of anaphylactogen and confirmation are still clinician to the judgement of patient of hypersensitivity's property disease and cognitive main means.The Allergic skin test mode applied clinically at present comprises detection and allergen detection in vitro in irritated substance.Detect in body and be mainly used in the judgement of anaphylactogen kind and the Preliminary detection of skin sensitivity, detection mode mainly comprises Skin-test (Histamine positive, intracutaneous test, scratch test, patch test, passive transfer test), provocative test (conjunctiva provocative test, Challenging Test On The Nasal Mucosa, BPT, oral provocative test); Vitro detection is mainly used in the judgement of cellular immunity and humoral immunity degree, and main detection mode comprises IgE coherent detection (specific IgE detection, total IgE detect) and cell activity assays (Determination of Tryptase, Basohil activation mensuration, eosinophil correlation factor measure).In addition, domesticly at present the detection mode of complete independently can be mainly Skin-test and IgE coherent detection, and Allergen provocation and Hypersensitivity Cells activity test not yet universal.The unification substantially of current vitro detection national and foreign standards, comprises the Uni-CAP system of goldstandard Phidia company, Fook company, Shanghai Ling Jian company etc.And the Allergic skin test mode world there is no goldstandard test in body, common external reagent comprise ALK company of Denmark peace thorn, German A Luoge company irritated substance in detection agent, French Stallergen company irritated substance in detection agent, but above offshore company only comprises Europe and U.S.'s Typical allergic former more than ten is planted, and does not comprise some Chinese distinctive anaphylactogen domestic.And domestic production producer is less, along with the incidence of anaphylactia raises year by year, and the sharply change of living environment, Typical allergic is former have been had with last century and has obviously changed.
Summary of the invention
The object of the present invention is to provide a kind of native allergens detection agent preparing instrument and application thereof, be intended to solve detection agent kind in irritated substance and make inconvenient problem very little, immediately.
The present invention is achieved in that a kind of native allergens detection agent preparing instrument, comprises loading slot, flux bath, low-temperature mixed pulverizing tank, low-temperature centrifugation extractor; Described loading slot top is provided with adding mouth, and bottom is provided with outlet, and described flux bath top is provided with solubilizer mouth, and bottom is provided with out solvent mouth;
The outlet of described loading slot, the solvent mouth that goes out of flux bath pulverize the inner conducting of tank respectively with low-temperature mixed, described low-temperature mixed pulverizes tank and low-temperature centrifugation extractor passes through connecting pipe conducting, this connecting pipe separates homogenate in tank is pulverized on road separatory outlet as low-temperature mixed; The top of described low-temperature centrifugation extractor is also provided with supernatant escape hole, and bottom is provided with sediment escape hole.
Preferably, described flux bath comprises the neutral flux groove for the acid flux material groove of splendid attire acid flux material, the basic solvent groove of splendid attire basic solvent and splendid attire neutral flux, and described acid flux material groove, basic solvent groove are identical with neutral flux groove structure.
Preferably, described native allergens detection agent preparing instrument also comprises some solenoid valves; Wherein, the outlet of described loading slot, acid flux material groove, basic solvent groove, neutral flux groove go out separatory outlet on solvent mouth, connecting pipe, the sediment escape hole of low-temperature centrifugation extractor is provided with solenoid valve respectively.
Preferably, described native allergens detection agent preparing instrument also comprises suction pumps; The inlet of described suction pumps is arranged in low-temperature centrifugation extractor supernatant layer, and the liquid outlet of described suction pumps is located at the outside of low-temperature centrifugation extractor.
Invention further provides above-mentioned native allergens detection agent preparing instrument detection agent in irritated substance prepare in application.
The present invention overcomes the deficiencies in the prior art, provides a kind of native allergens detection agent preparing instrument, comprises loading slot, flux bath, low-temperature mixed pulverizing tank, low-temperature centrifugation extractor; Loading slot top is provided with adding mouth, and bottom is provided with outlet, and flux bath top is provided with solubilizer mouth, and bottom is provided with out solvent mouth; The outlet of loading slot, the solvent mouth that goes out of flux bath pulverize the inner conducting of tank respectively with low-temperature mixed, low-temperature mixed pulverizes tank and low-temperature centrifugation extractor passes through connecting pipe conducting, this connecting pipe separates homogenate in tank is pulverized on road separatory outlet as low-temperature mixed; The top of low-temperature centrifugation extractor is also provided with supernatant escape hole, and bottom is provided with sediment escape hole.
The present invention's application natural materials, the mycotic spore that the pollen of such as fresh food, fresh collection, local are collected, the compound (restodent) etc. suspecting allergy, by pulverizing, mixing, centrifugal, extraction, be separated, the mode such as quantitative is prepared into the anaphylactogen immersion liquid reagent that can be used for detecting in Clinical allergy substance, directly can the pricking method method of use safety specification carry out detecting whether to this type of allergies.
Compared to the shortcoming and defect of prior art, the present invention has following beneficial effect:
(1) not by the restriction of anaphylactogen kind, the allergin of clinical signs of suspected scope all can use this equipment to be prepared as reagent for medical treatment.
(2) preparation time is of short duration, can keep the biologically active of allergenic agent, avoids the interference of other materials (easily having the growth of mould in dirt mite).
(3) cheap, preparation process only needs to collect a certain amount of suspection material, without the need to commercial reagents, with low cost.
(4) calculating of volume weight ratio can be carried out, carry out semi-quantitative analysis, be conducive to the assessment of allergic degree.
(5) different component separating and extracting is carried out to allergin, the possibility of rare allergenic components allergy (be mainly the molten wheat component allergy of alcohol as wheat relies on exercise induced type shock, and Wheat Dood Allergy stoste being mainly water-soluble) can be investigated.
(6) anaphylactogen is passed for gas to detect according to the peculiar allergin in locality (such as camphor tree allergy there is no research).
(7) detection can be carried out with food after original food or culinary art for food allergen and keep anaphylactogen integrality and promptness.
(8) pricking method or patch detection is carried out after can directly using compound finished product to collect mixing for professional anaphylactogen, assess whether after allergy again row composition allergy detect (such as which kind of hair dye is hair dye allergy can first investigate as, and " Invest, Then Investigate " Allergen).
(9) different reagent Omnibus Research incompatibility and irritated component analysis (allergic reaction of Chinese medicine preparation judges) can be carried out for special anaphylactogen as medicine or biopreparate, because concentration height is controlled, the bad reaction of common skin test can be avoided.
(10) can be used for basic medical unit, equipment price is cheap, and anaphylactogen preparation method is simple, for without commercialization or region Allergic skin test with the obvious advantage.
(11) for external conventional point-to-point pricking method mode, owing to using food original matter likely to cause excessive concentration to cause bad reaction, and this equipment adopts the semiquantitative way of w/v can effectively reduce this type of risk.
Accompanying drawing explanation
Fig. 1 is the structural representation of native allergens detection agent preparing instrument of the present invention.
Embodiment
In order to make object of the present invention, technical scheme and advantage clearly understand, below in conjunction with drawings and Examples, the present invention is further elaborated.Should be appreciated that specific embodiment described herein only in order to explain the present invention, be not intended to limit the present invention.
As shown in Figure 1, Fig. 1 is the structural representation of native allergens detection agent preparing instrument of the present invention.
A kind of native allergens detection agent preparing instrument, comprise loading slot 1, flux bath (described flux bath comprises the neutral flux groove 4 for the acid flux material groove 2 of splendid attire acid flux material, the basic solvent groove 3 of splendid attire basic solvent and splendid attire neutral flux, and described acid flux material groove 2, basic solvent groove 3 are identical with neutral flux groove 4 structure), low-temperature mixed pulverizing tank 5, low-temperature centrifugation extractor 6; Described loading slot 1 top is provided with adding mouth, and bottom is provided with outlet; Described acid flux material groove 2, basic solvent groove 3 and neutral flux groove 4 top are respectively equipped with solubilizer mouth, and bottom is respectively equipped with out solvent mouth;
The solvent mouth that goes out of the outlet of described loading slot 1, acid flux material groove 2, basic solvent groove 3 and neutral flux groove 4 pulverizes the inner conducting of tank 5 with low-temperature mixed respectively, described low-temperature mixed is pulverized tank 5 and is passed through connecting pipe 7 conducting with low-temperature centrifugation extractor 6, this connecting pipe 7 separates homogenate in tank 5 is pulverized on road separatory outlet as low-temperature mixed; The top of described low-temperature centrifugation extractor 5 is also provided with supernatant escape hole 8, and bottom is provided with sediment escape hole 9.
In embodiments of the present invention, more specifically, described native allergens detection agent preparing instrument also comprises some solenoid valves; Wherein, the outlet of described loading slot 1, acid flux material groove 2, basic solvent groove 3, neutral flux groove 4 go out separatory outlet on solvent mouth, connecting pipe 7, the sediment escape hole of low-temperature centrifugation extractor 6 is provided with solenoid valve respectively.
In the present invention, more specifically, described native allergens detection agent preparing instrument also comprises suction pumps (omitting view in figure); Described suction pumps is located in described low-temperature centrifugation extractor 6, and the inlet of described suction pumps is arranged in low-temperature centrifugation extractor 6 supernatant layer, and the liquid outlet of described suction pumps is located at the outside of low-temperature centrifugation extractor 6.
In the present invention, more specifically, described low-temperature mixed pulverizes tank 5 can with the comminuting method of two kinds of principles, the first is that directly pulverizing is stirred in application, there are two wells (control element can be utilized to be rotated counterclockwise) above right cylinder closed container, add sample after a hole is rotated in place, after another hole is rotated in place, add solvent, well closed by closed container afterwards, utilizes to stir to pulverize (such as adding with juice extractor, comminutor).The second utilizes sonioation method, also be first application of sample and add solvent and then utilize ultrasonication (at present conventional Ultrasonic Cell Disruptor has the JY92-IIDN type Ultrasonic Cell Disruptor of SCIENTZ board), because crushing process can produce high temperature, the protein active of test substance may be destroyed, therefore add low temperature in a reservoir and maintain equipment, such as controlled refrigerating plant.And low-temperature centrifugation extractor 6 essence is refrigerated centrifuge.
In embodiments of the present invention, above-mentioned solenoid valve, low-temperature mixed are pulverized tank 5, low-temperature centrifugation extractor 6, suction pumps and are all arranged Automated condtrol element controlled work by surface-mounted integrated circuit.
The course of work of native allergens detection agent preparing instrument of the present invention specifically comprises the following steps:
1, add testing sample in loading slot 1, in acid flux material groove 2, basic solvent groove 3 and neutral flux groove 4, add corresponding reagent respectively;
2, low-temperature mixed pulverizes tank 5 is counterclockwise controllability device, according to thinking that requirement adds testing sample and mixed solvent successively, carries out low-temperature mixed pulverizing afterwards by Automated condtrol element;
3, after co-grinding, sample is collected by homogenate outlet or directly enters low-temperature centrifugation extractor 6;
4, compounding substances starts Centrifugical extraction according to after Automated condtrol element adjustment rotating speed and time setting in low-temperature centrifugation extractor 6;
5, low-temperature centrifugation extractor 6 stops centrifugal work, and suction pump passes through the supernatant escape hole 8 sucking-off supernatant of low-temperature centrifugation extractor 6, by sediment escape hole 9 collecting precipitation protein.
Native allergens detection agent preparing instrument application natural materials of the present invention, the mycotic spore that the pollen of such as fresh food, fresh collection, local are collected, the compound (restodent) etc. suspecting allergy, by pulverizing, mixing, centrifugal, extraction, be separated, the mode such as quantitative is prepared into the anaphylactogen immersion liquid reagent that can be used for detecting in Clinical allergy substance, directly can the pricking method method of use safety specification carry out detecting whether to this type of allergies.
The foregoing is only preferred embodiment of the present invention, not in order to limit the present invention, all any amendments done within the spirit and principles in the present invention, equivalent replacement and improvement etc., all should be included within protection scope of the present invention.
Claims (5)
1. a native allergens detection agent preparing instrument, is characterized in that, comprises loading slot, flux bath, low-temperature mixed pulverizing tank, low-temperature centrifugation extractor; Described loading slot top is provided with adding mouth, and bottom is provided with outlet, and described flux bath top is provided with solubilizer mouth, and bottom is provided with out solvent mouth;
The outlet of described loading slot, the solvent mouth that goes out of flux bath pulverize the inner conducting of tank respectively with low-temperature mixed, described low-temperature mixed pulverizes tank and low-temperature centrifugation extractor passes through connecting pipe conducting, this connecting pipe separates homogenate in tank is pulverized on road separatory outlet as low-temperature mixed; The top of described low-temperature centrifugation extractor is also provided with supernatant escape hole, and bottom is provided with sediment escape hole.
2. native allergens detection agent preparing instrument as claimed in claim 1, it is characterized in that, described flux bath comprises the neutral flux groove for the acid flux material groove of splendid attire acid flux material, the basic solvent groove of splendid attire basic solvent and splendid attire neutral flux, and described acid flux material groove, basic solvent groove are identical with neutral flux groove structure.
3. native allergens detection agent preparing instrument as claimed in claim 2, it is characterized in that, described native allergens detection agent preparing instrument also comprises some solenoid valves; Wherein, the outlet of described loading slot, acid flux material groove, basic solvent groove, neutral flux groove go out separatory outlet on solvent mouth, connecting pipe, the sediment escape hole of low-temperature centrifugation extractor is provided with solenoid valve respectively.
4. native allergens detection agent preparing instrument as claimed in claim 3, it is characterized in that, described native allergens detection agent preparing instrument also comprises suction pumps; The inlet of described suction pumps is arranged in low-temperature centrifugation extractor supernatant layer, and the liquid outlet of described suction pumps is located at the outside of low-temperature centrifugation extractor.
5. the native allergens detection agent preparing instrument detection agent in irritated substance described in Claims 1 to 4 prepare in application.
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RU2304000C1 (en) * | 2006-02-10 | 2007-08-10 | Государственное учреждение Научно-исследовательский институт вакцин и сывороток им. И.И. Мечникова Российской академии медицинских наук | Method for producing diagnostic allergen |
CN102109431A (en) * | 2010-12-21 | 2011-06-29 | 江南大学 | Method for preparing shrimp allergen standard substance |
CN103645328A (en) * | 2013-12-17 | 2014-03-19 | 中国计量学院 | Method for preparing peanut allergic protein Arah2 standard |
CN103822999A (en) * | 2014-02-28 | 2014-05-28 | 中国农业科学院农产品加工研究所 | Full-automatic QuEChERS preprocessing all-in-one machine and preprocessing method |
CN204374176U (en) * | 2015-01-26 | 2015-06-03 | 华中科技大学同济医学院附属同济医院 | Natural allergen detection agent preparation instrument |
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