CN114504659A

CN114504659A - Nasal cavity excitation kit and preparation method and application thereof

Info

Publication number: CN114504659A
Application number: CN202210111934.7A
Authority: CN
Inventors: 叶菁; 沈李
Original assignee: Individual
Current assignee: Individual
Priority date: 2022-01-29
Filing date: 2022-01-29
Publication date: 2022-05-17
Anticipated expiration: 2042-01-29
Also published as: CN114504659B

Abstract

The invention discloses a nasal cavity excitation kit, a preparation method and application, wherein the nasal cavity excitation kit comprises: a nasal cavity stimulating spray comprising a nasal cavity stimulating agent comprising an allergen; a solvent spray, which contains a solvent. The kit comprises a nasal cavity excitation spraying agent and a solvent spraying agent, when the kit is used for a nasal cavity allergen excitation test, the allergen can be constantly, repeatedly and uniformly distributed in a nasal cavity mucous membrane, the allergen excitation process under the natural environment can be simulated as far as possible, and the kit has the advantages of safety, economy, simplicity in operation, rapidness, convenience, high repeatability, high patient acceptance and easiness in popularization.

Description

Nasal cavity excitation kit and preparation method and application thereof

Technical Field

The invention belongs to the technical field of biological preparations, and particularly relates to a nasal cavity excitation kit, and a preparation method and application thereof.

Background

Allergic Rhinitis (AR) is an Immunoglobulin E (IgE) -mediated type I hypersensitivity to allergens, affecting 10-40% of the world population^[1]. The incidence rate of AR in the main cities of China is 4 to 38 percent^[2]. Clinical symptoms such as nasal obstruction, watery nasal discharge, nasal itching, sneezing, etc. in combination with allergen Skin Prick Test (SPT) and/or serum allergen Specific IgE (sIgE) are the clinical routine methods for diagnosing AR^[1]. SPT and serum sIgE correspond to systemic allergic inflammatory response, only can reflect the expression condition of sIgE in vivo, and cannot necessarily reflect the local pathophysiology and physiology of nasal mucosa^[3]. Non-allergic rhinitis (NAR) patients also often show sneezing, watery nasal discharge, nasal congestion and nasal itching, which are highly similar to the symptoms of allergic rhinitis, but the NAR patient allergen SPNegative results of T and serum sIgE examination^[4]This group of patients currently lacks a clinically definitive diagnostic method. In addition, it has been found that some of AR patients have type I hypersensitivity reactions only in the nasal mucosa, producing local sIgE^[5]. These locally expressed sIgE in the nasal mucosa cannot be detected by serum sIgE or SPT examination. These patients are defined as Local Allergic Rhinitis (LAR). Patients diagnosed with NAR by examination of allergen SPT and serum sIgE, after performing Nasal allergen challenge test (NAPT), 26.5% of NAR patients were confirmed as LAR^[6]. To identify AR, NAR and LAR, NAPT is required.

NAPT is the internationally recognized gold standard for diagnosing AR^[7]Can objectively reflect allergic reaction of nasal mucosa. NAPT acts specific allergens on the nasal mucosa in a quantitative manner in order to induce type i hypersensitivity of the nasal mucosa, diagnoses AR and determines the allergens of AR patients by evaluating the clinical symptoms and signs of the nose before and after challenge within a specific time. NAPT results are determined by subjective scoring and/or objective evaluation methods. The subjective scoring system includes Likert score, Total Nasal Symptoms Score (TNSS), VAS score, Linder score, and Lebel score. Objective evaluation systems include Peak Nasal Inspiratory Flow (PNIF), nasal acoustic reflex measurements, active anterior nasal tonometry, and four-phase nasal resistance testing. Although the prior studies have adopted different test schemes and result evaluation methods, no international uniform standard has been achieved for the normalization of NAPT. But the European Association of allergy and clinical immunology^[8]The methods and conclusions of all the above studies are approved.

Allergen Immunotherapy (AIT) of AR is currently the only recognized causative treatment that can alter the natural course of allergy. Without NAPT, LAR patients cannot be diagnosed, missing AIT treatment. NAPT can also be used to validate allergens before attempting AIT in AR patients who have been positively diagnosed with SPT or serum sIgE results. It is also important that AR patients will achieve stable and long lasting efficacy after receiving about 3 years of continuous therapy, but studies have been conducted^[9]Indicating that patients who still partially received AIT failed to obtain the desired therapeutic effect. This may be because such patients are in a "primed" (SPT or serum sIgE test positive, NAPT test negative) state, which makes it difficult for such patients to benefit from AIT. This highlights the importance of NAPT.

At present, a related research report of NAPT implementation is lacked in China. In foreign research reports, NAPT may be implemented using a dropping method, a filter paper method, a spraying method, and an allergen challenge chamber. Wherein the dripping method comprises dripping allergen solution on the mucosa of inferior turbinate by use of injector, dropper or micropipette. Although the method is easy to operate, the allergen distribution range is unpredictable and uneven, and the allergen may be deposited in the pharynx, larynx and bronchus, and even the risk of inducing bronchial asthma attack and causing severe anaphylaxis exists. The filter paper method is to apply the filter paper soaked with allergen to mucosa of middle turbinate and inferior turbinate. Although the method can save the using amount of the allergen reagent, the filter paper sheet can not be uniformly spread in the nasal cavity due to nasal cavity swelling and stenosis, and the operation can also cause nasal cavity pain of a patient, and has the risks of causing nasal mucosa injury, epistaxis and sinus disease induction. The existing spray method nasal cavity excitation test has the following problems: 1. the allergen is unevenly distributed in the nasal cavity during the use process, and the reasons include unstable and uneven dispersion of the allergen in the nasal cavity stimulating agent and uneven and consistent spray particles during the use; 2. the pressure and spray stroke of a spray bottle used in the spray method are not constant and inconsistent, and the nasal cavity stimulating reagent cannot be quantitatively sprayed out; 3. under the condition of not setting gradient concentration, the test cannot reflect the nasal cavity hyperreactivity degree of patients with different diseases. In addition, allergen challenge compartments are currently recognized devices or equipment that may be used in NAPTs. Allergen excitation chambers (EECs) are one of the closest approaches to daily allergen exposure, and the safety, effectiveness, and repeatability of this approach are currently proven. EEC is a controlled exposure of patients to allergens in a closed environment, where the allergens are released by specific means in the room and the concentration of allergens in the air can be monitored and adjusted in real time. The device can achieve the effect of simulating the distribution characteristics of the allergen in the natural environment so as to closely reproduce the natural allergen nasal cavity excitation process. However, EECs are complex to operate and extremely expensive in equipment, which can cost as high as tens of millions. The number of EECs established worldwide is very limited, 1 EEC is owned in Japan in Asia range, and the EEC has no unified excitation standard and is not popularized at present.

Therefore, no kit product related to the nasal cavity excitation test is available in the market at present, so as to solve the problems of insecurity, high price, complex operation, low repeatability, low patient acceptance, difficulty in popularization and use and the like of the existing method, reagent, material or device for the nasal cavity excitation test.

The relevant documents are given below:

[1]Wang XD,Zheng M,Lou HF,et al.An increased prevalence of self-reported allergic rhinitis in major Chinese cities from 2005to 2011.Allergy 2016；71(8):1170-1180.

[2]Brozek JL,Bousquet J,Agache I,et al.Allergic Rhinitis and its Impact on Asthma(ARIA)guidelines-2016revision.J Allergy Clin Immunol 2017；140(4):950-958.

[3]Powe DG,Bonnin AJ,Jones NS.'Entopy':local allergy paradigm.Clin Exp Allergy 2010；40(7):987-997.

[4]Rondon C,Bogas G,Barrionuevo E,et al.Nonallergic rhinitis and lower airway disease.Allergy 2017；72(1):24-34.

[5]Gosepath J,Amedee RG,Mann WJ.Nasal provocation testing as an international standard for evaluation of allergic and nonallergic rhinitis.Laryngoscope 2005；115(3):512-516.

[6]Auge J,Vent J,Agache I,et al.EAACI Position paper on the standardization of nasal allergen challenges.Allergy 2018；73(8):1597-1608.

[7]Shamji MH,Kappen JH,Akdis M,et al.Biomarkers for monitoring clinical efficacy of allergen immunotherapy for allergic rhinoconjunctivitis and allergic asthma:an EAACI Position Paper.Allergy 2017；72(8):1156-1173.

[8]Hamizan AW,Rimmer J,Alvarado R,et al.Positive allergen reaction in allergic and nonallergic rhinitis:a systematic review.Int Forum Allergy Rhinol 2017；7(9):868-877.

[9]Gould HJ,Ramadani F.IgE responses in mouse and man and the persistence of IgE memory.Trends Immunol 2015；36(1):40-48.

disclosure of Invention

The invention aims to solve the technical problems that a kit product related to a nasal cavity excitation test does not appear in the prior art, and the existing method, reagent, material or device for the nasal cavity excitation test is unsafe, high in price, complex in operation, low in repeatability, low in patient acceptance, difficult to popularize and use and the like.

In order to solve the above technical problem, an embodiment of the present invention provides a nasal cavity excitation kit, including:

a nasal cavity stimulating spray comprising a nasal cavity stimulating agent comprising an allergen;

a solvent spray, which contains a solvent.

Optionally, the nasal cavity stimulating spray can uniformly and quantitatively spray the nasal cavity stimulating reagent, and the solvent spray can uniformly and quantitatively spray the solvent.

Alternatively, the nasal cavity stimulating spray may contain only a single type of allergen per unit size, the allergen being an inhalant allergen.

Alternatively, the allergen includes, but is not limited to, a dermatophagoides farinae allergen, a dermatophagoides pteronyssinus allergen, a ragweed allergen, an alternaria allergen, a dog hair allergen, a cat hair allergen, a phoenix tree allergen, a birch allergen, a grass pollen allergen, a ragweed pollen allergen, a tropical mite allergen, a blattaria germanica allergen, an alternaria allergen, an aspergillus niger allergen, an mugwort allergen, a phoenix tree allergen, a willow allergen, a poplar allergen, a birch allergen, a hop allergen, an elm pollen allergen, or a cedar pollen allergen.

Optionally, the solvent comprises one or more of phenol, sodium dihydrogen phosphate, disodium phosphate, sodium oxide, glycerol, and water for injection.

Optionally, the nasal cavity stimulating agent and the solvent are respectively filled in a spray bottle, a drug spray pump is arranged on the spray bottle, and each spray of the nasal cavity stimulating agent and the solvent spray is 0.04mL to 0.06 mL.

Optionally, the nasal cavity excitation reagent comprises a nasal cavity excitation reagent stock solution with gradient concentration, and the nasal cavity excitation reagent stock solution is dermatophagoides pteronyssinus allergen prick liquid.

Optionally, the concentration of allergen in the nasal cavity challenge reagent stock solution is from 8HEP to 10 HEP.

The nasal cavity stimulating reagent stock solution with gradient concentration comprises: 1/1000 nasal cavity stimulating agent stock solution, 1/100 nasal cavity stimulating agent stock solution, 1/10 nasal cavity stimulating agent stock solution and nasal cavity stimulating agent stock solution.

The embodiment of the invention also provides a preparation method of the nasal cavity stimulation kit, which is used for preparing the nasal cavity stimulation kit and comprises the following steps:

preparing a solvent;

preparing a nasal cavity stimulating reagent;

and respectively filling the solvent and the nasal cavity excitation reagent into spray bottles to obtain the nasal cavity excitation kit.

Optionally, the preparation method comprises:

preparing a solvent, and filling 1000 parts by volume of the solvent into a spray bottle to prepare 1000 parts by volume of the solvent spray;

preparing a nasal cavity stimulating reagent, wherein the nasal cavity stimulating reagent comprises a nasal cavity stimulating reagent stock solution, taking four spray bottles, respectively filling 1 volume part of the nasal cavity stimulating reagent stock solution, 999 volume parts of a solvent, 10 volume parts of the nasal cavity stimulating reagent stock solution, 990 volume parts of a solvent, 100 volume parts of the nasal cavity stimulating reagent stock solution, 900 volume parts of the solvent and 1000 volume parts of the nasal cavity stimulating reagent stock solution into the four spray bottles, and respectively obtaining 1000 volume parts of the nasal cavity stimulating spray, thus obtaining the nasal cavity stimulating kit.

The embodiment of the invention also provides application of the nasal cavity excitation kit in a nasal cavity allergen excitation test, wherein the nasal cavity excitation kit is the nasal cavity excitation kit.

Optionally, the use of the nasal cavity challenge kit in a nasal cavity allergen challenge test comprises:

filling out a basic rhinitis symptom VAS score, and taking the score as a reference basic value;

using the solvent spray to carry out a solvent excitation test, if the VAS score increases relative to the basic value and is less than the reference value, judging that the result is negative, and carrying out an allergen excitation test, otherwise, judging that the result is positive, and carrying out the solvent excitation test after two weeks;

performing allergen excitation test by using the nasal cavity excitation spraying agent, if the VAS score increases more than or equal to a reference value relative to a basic value, judging that the excitation test result is positive, ending the inspection, and recording the concentration threshold of the nasal cavity excitation reagent; otherwise, it is "negative".

Optionally, the reference value is set to 55 points.

carrying out a solvent excitation test by using the solvent spray, spraying 1 spray to the nasal cavities at two sides of a patient by using the solvent spray, filling a VAS score of rhinitis symptoms immediately after 10 minutes, if the VAS score is increased by less than 55 points relative to a basic value, judging that the result is negative, carrying out an allergen excitation test, and if the result is positive, carrying out the solvent excitation test after two weeks;

performing allergen excitation test by using the nasal cavity excitation spraying agent, spraying 1 spray to the nasal cavities at both sides of a patient by using the nasal cavity excitation spraying agent containing 1/1000 concentration nasal cavity excitation reagent stock solution, filling the VAS score of rhinitis symptoms immediately after 10 minutes, judging that the excitation test result is positive if the VAS score is increased by more than or equal to 55 points relative to a basic value, finishing the inspection, and recording the concentration threshold value of the nasal cavity excitation reagent; otherwise, the result is negative, and the inspection A is required;

checking A: spraying 1 spray of nasal cavity stimulating spray containing nasal cavity stimulating reagent stock solution with 1/100 concentration into nasal cavities of both sides of a patient respectively, filling out a VAS score of rhinitis symptoms immediately after 10 minutes, judging that a stimulating test result is positive if the VAS score is increased by more than or equal to 55 points relative to a basic value, finishing the inspection, and recording a concentration threshold value of the nasal cavity stimulating reagent; otherwise, the test is negative, and a test B is required;

and B, checking: spraying 1 spray of nasal cavity stimulating spray containing nasal cavity stimulating reagent stock solution with 1/10 concentration into nasal cavities of both sides of a patient respectively, filling out a VAS score of rhinitis symptoms immediately after 10 minutes, judging that a stimulating test result is positive if the VAS score is increased by more than or equal to 55 points relative to a basic value, finishing the inspection, and recording a concentration threshold value of the nasal cavity stimulating reagent; otherwise, the result is negative, and the test C needs to be carried out;

and (5) checking C: spraying 1 spray of a nasal cavity stimulating spray containing a nasal cavity stimulating reagent stock solution to the nasal cavities on the two sides of a patient respectively, filling a VAS score of rhinitis symptoms immediately after 10 minutes, judging that a stimulating test result is positive if the VAS score is increased by more than or equal to 55 points relative to a basic value, and recording a nasal cavity stimulating reagent concentration threshold; otherwise, judging the result of the patient arousal test as negative.

The embodiment of the invention also provides application of the house dust mite allergen pricking liquid in a nasal cavity allergen excitation test, wherein the house dust mite allergen pricking liquid is derived from a mite allergen skin pricking kit, the mite allergen skin pricking kit is named as Anci in Chinese, Soluprick SQ in English, and ALK-Abello A/S is a manufacturer.

Compared with the prior art, the invention has the following advantages:

the nasal cavity excitation kit provided by the embodiment of the invention comprises the nasal cavity excitation spray and the solvent spray, when the kit is used for a nasal cavity allergen excitation test, not only can the allergen be constantly, repeatedly and uniformly distributed in a nasal cavity mucous membrane, and the allergen excitation process under the natural environment can be simulated as much as possible, but also the kit has the advantages of safety, economy, simplicity in operation, rapidness, convenience, high repeatability, high patient acceptance and easiness in popularization.

Further, be provided with medicine spraying agent pump on the spray bottle, nasal cavity arouses spraying agent and menstruum spraying agent each to spout for 0.04mL to 0.06mL, the nasal cavity arouses spraying agent can be even, the ration blowout nasal cavity arouses reagent, menstruum spraying agent can be even, the ration blowout the menstruum, when specifically carrying out the nasal cavity arouses the experiment, only need each nostril of every side respectively spout 1 time, can make the allergen spray evenly cover on well turbinate and inferior turbinate mucous membrane, not only the security of kit is high, and economic convenience, patient's acceptance is high, can not cause the obvious discomfort of patient.

Further, the nasal cavity stimulating reagent provided by the embodiment of the invention is a nasal cavity stimulating reagent stock solution with gradient concentration, the nasal cavity stimulating reagent is set through gradient concentration, discomfort of patients caused by direct detection of overhigh concentration of the nasal cavity stimulating reagent is avoided, the nasal cavity hyperreactivity degree of patients with different diseases can be reflected through NAPT test, AR, NAR and LAR are detected according to the nasal cavity allergen stimulating test, and reference values are provided for helping to judge the type of allergic rhinitis and whether AIT treatment can be carried out or not or predicting AIT treatment effect.

In addition, the embodiment of the invention also provides a preparation method of the nasal cavity excitation kit, and the preparation method of the nasal cavity excitation kit is simple and convenient to popularize and apply.

The embodiment of the invention also provides application of the nasal cavity excitation kit in a nasal cavity allergen excitation test, and the nasal cavity allergen excitation test performed by using the nasal cavity excitation kit is safe, economic, simple to operate, high in repeatability, high in patient acceptance and easy to popularize.

The embodiment of the invention also provides application of the dermatophagoides pteronyssinus allergen prick liquid in a nasal cavity allergen excitation test, widens the application of the dermatophagoides pteronyssinus allergen prick liquid, and provides a standard reagent which is safe, economical, simple to operate, high in repeatability, high in patient acceptance and easy to popularize for a nasal cavity excitation test by a spray method.

Drawings

Fig. 1 is a process flow diagram of a method of making a nasal cavity challenge kit according to some embodiments of the present invention.

Fig. 2 is a process flow diagram of a method of making a nasal cavity challenge kit according to further embodiments of the present invention.

Fig. 3 is a VAS sample table in an example of the present invention.

Fig. 4 is a flow chart of the application of the nasal cavity excitation kit in the nasal cavity allergen excitation test according to the embodiment of the invention.

Detailed Description

As described in the background, NAPT is the internationally accepted gold standard for diagnosis of AR, and it is currently practiced using the drip method, filter paper method, spray method, and allergen challenge chamber. The method, the reagent, the material or the device for the nasal cavity excitation test have the problems of unsafe use, high price, complex operation, low repeatability, low patient acceptance, difficult popularization and use and the like at present, and a kit product related to the nasal cavity excitation test for solving the problems does not exist in the market at present.

The nasal cavity excitation kit provided by the embodiment of the invention comprises the nasal cavity excitation spray and the solvent spray, when the kit is used for a nasal cavity allergen excitation test, not only can the allergen be constantly, repeatedly and uniformly distributed in a nasal cavity mucous membrane, but also the allergen excitation process under the natural environment can be simulated as far as possible, and the kit has the advantages of safety, economy, simplicity in operation, rapidness, convenience, high repeatability, high patient acceptance and easiness in popularization.

The embodiment of the invention also provides a preparation method of the nasal cavity excitation kit, and the preparation method of the nasal cavity excitation kit is simple and convenient to popularize and apply.

The embodiment of the invention also provides application of the house dust mite allergen pricking liquid in a nasal cavity allergen excitation test, widens the application of the house dust mite allergen pricking liquid, and provides a standard reagent which is safe, economical, simple to operate, high in repeatability, high in patient acceptance and easy to popularize for the nasal cavity excitation test by a spraying method.

In order to make the aforementioned objects, features and advantages of the present invention comprehensible, embodiments accompanied with figures are described in detail below.

In the present invention, the source of the reagents, apparatuses and equipment used is not particularly limited unless otherwise specified, and commercially available products, for example, may be used.

The embodiment of the invention provides a nasal cavity excitation kit, which comprises:

a solvent spray, which contains a solvent.

Wherein the allergen contained in the nasal cavity stimulating reagent is used for inducing nasal mucosa hypersensitivity. In some embodiments, the nasal excitation spray contains only a single species of allergen per unit size, which is an inhalant allergen.

In some embodiments, the allergen includes, but is not limited to, a dermatophagoides farinae allergen, a dermatophagoides pteronyssinus allergen, a ragweed allergen, an alternaria allergen, a dog hair allergen, a cat hair allergen, a phoenix tree allergen, a birch allergen, a grass pollen allergen, a ragweed pollen allergen, a tropical mite allergen, a blattaria germanica allergen, an alternaria alternata allergen, an aspergillus niger allergen, an artemisia argyi allergen, a phoenix tree allergen, a willow allergen, a poplar allergen, a birch allergen, a hop allergen, an elm pollen allergen, or a cedar pollen allergen.

Wherein the solvent is a liquid for dissolving or dispersing the allergen. The solvent comprises one or more of phenol, sodium dihydrogen phosphate, disodium phosphate, sodium oxide, glycerol or water for injection. In general, the choice of the vehicle in the present invention is not particularly limited as long as the allergen can be dissolved or dispersed. For example, in one embodiment, the vehicle is formulated from phenol, sodium dihydrogen phosphate, disodium phosphate, sodium oxide, glycerol, and water for injection.

In some embodiments, the nasal cavity stimulating agent and the vehicle are separately contained in a spray bottle having a drug spray pump, and the nasal cavity stimulating spray and the vehicle spray are each 0.04 to 0.06 mL. The medicine spray pump arranged on the spray bottle can ensure that the nasal cavity excitation spray can uniformly and quantitatively spray the nasal cavity excitation reagent and the solvent spray can uniformly and quantitatively spray the solvent. For example, in one embodiment, the nasal cavity stimulating spray and vehicle spray are 0.05mL each. When the nasal cavity excitation test is specifically carried out, only 1 time of spraying is needed to be carried out on each nostril, so that the allergen spray can uniformly cover the mucous membranes of the middle turbinate and the lower turbinate, the safety of the kit is high, the kit is economical and convenient, the acceptance of a patient is high, and the obvious discomfort of the patient can not be caused.

In some embodiments, the nasal cavity stimulating reagent comprises a gradient concentration of a nasal cavity stimulating reagent stock solution, the nasal cavity stimulating reagent stock solution being a dermatophagoides pteronyssinus allergen prick solution. In some embodiments, the concentration of allergen in the nasal challenge reagent stock is from 8HEP to 10 HEP. For example, in one embodiment, the nasal cavity challenge reagent stock has a concentration of 10 HEP. It should be noted that, in general, the gradient concentration setting of the nasal cavity stimulating agent is not particularly limited in the present invention, as long as the discomfort of the patient caused by the unreasonable concentration gradient setting can be avoided, and it is ensured that the measured VAS score can be used to determine the allergic status of the patient with different disease conditions. For example, in one embodiment, the gradient concentration nasal cavity stimulating reagent stock comprises: 1/1000 nasal cavity stimulating agent stock solution, 1/100 nasal cavity stimulating agent stock solution, 1/10 nasal cavity stimulating agent stock solution and nasal cavity stimulating agent stock solution. The nasal cavity stimulating reagent is arranged through gradient concentration, discomfort of a patient caused by direct detection of overhigh concentration of the nasal cavity stimulating reagent is avoided, the nasal cavity hyperreactivity degree of different patients with different disease states can be reflected through NAPT test, AR, NAR and LAR are detected according to the nasal cavity allergen stimulating test, and the reference value is provided for helping to judge the type of allergic rhinitis and whether AIT treatment can be carried out or not or predicting the AIT treatment effect.

Fig. 1 is a process flow diagram of a method of making a nasal cavity challenge kit according to some embodiments of the present invention. The preparation method of the nasal cavity stimulation kit according to the embodiment of the present invention can be applied to the preparation of the nasal cavity stimulation kit according to the embodiment of the present invention, and the preparation method is shown in each step in fig. 1.

In these embodiments, the preparation method comprises:

s10, preparing a solvent;

s20, preparing a nasal cavity stimulating reagent;

s30, respectively filling the solvent and the nasal cavity stimulating reagent into spray bottles to obtain the nasal cavity stimulating kit.

Fig. 2 is a process flow diagram of a method of making a nasal cavity challenge kit according to further embodiments of the present invention. The preparation method of the nasal cavity stimulation kit provided by the embodiment of the invention can be applied to the preparation of the nasal cavity stimulation kit provided by the embodiment of the invention, namely the steps in fig. 2.

In these embodiments, the preparation method comprises:

s100, preparing a solvent, and filling 1000 parts by volume of the solvent into a spray bottle to prepare 1000 parts by volume of a solvent spray;

s200, preparing a nasal cavity stimulating reagent, wherein the nasal cavity stimulating reagent comprises a nasal cavity stimulating reagent stock solution, taking four spray bottles, respectively filling 1 volume part of the nasal cavity stimulating reagent stock solution, 999 volume parts of a solvent, 10 volume parts of the nasal cavity stimulating reagent stock solution, 990 volume parts of the solvent, 100 volume parts of the nasal cavity stimulating reagent stock solution, 900 volume parts of the solvent and 1000 volume parts of the nasal cavity stimulating reagent stock solution into the four spray bottles, and respectively obtaining 1000 volume parts of the nasal cavity stimulating spray, thus obtaining the nasal cavity stimulating kit.

It can be understood that the nasal cavity stimulating kit prepared by the above preparation method comprises 1 bottle of 1000 parts by volume of the solvent spray and four bottles of 1000 parts by volume of the nasal cavity stimulating spray, wherein the four bottles of nasal cavity stimulating agents comprise a gradient concentration nasal cavity stimulating agent stock solution, and the gradient concentration nasal cavity stimulating agent stock solution is: 1/1000 nasal cavity stimulating reagent stock solution, 1/100 nasal cavity stimulating reagent stock solution, 1/10 nasal cavity stimulating reagent stock solution and nasal cavity stimulating reagent stock solution. In addition, during actual preparation, nasal cavity stimulating kits with different specifications can be prepared according to requirements. For example, in one embodiment, the formulation is formulated to a size of 2 mL/vial to provide a kit with a size of 2mL nasal and vehicle sprays per vial. The preparation method for preparing the nasal cavity excitation kit is simple and convenient to popularize and apply.

The embodiment of the invention also provides application of the nasal cavity excitation kit in a nasal cavity allergen excitation test, wherein the nasal cavity excitation kit is the nasal cavity excitation kit disclosed by the embodiment of the invention, and the nasal cavity allergen excitation test is performed by using the nasal cavity excitation kit disclosed by the invention, so that the nasal cavity allergen excitation test is safe, economic, simple to operate, high in repeatability, high in patient acceptance and easy to popularize.

In some embodiments, the use of the nasal cavity challenge kit in a nasal cavity allergen challenge assay comprises:

In some embodiments, the reference value is set to 55 points. The reference value is set to 55 points, so that the result of the excitation test can be accurately judged to be negative or positive.

In other embodiments, the use of the nasal cavity challenge kit in a nasal cavity allergen challenge assay comprises:

performing allergen excitation test by using the nasal cavity excitation spraying agent, spraying 1 spray to the nasal cavities at both sides of a patient by using the nasal cavity excitation spraying agent containing '1/1000 concentration nasal cavity excitation reagent stock solution', filling out the VAS score of rhinitis symptoms immediately after 10 minutes, judging that the excitation test result is positive if the VAS score is increased by more than or equal to 55 points relative to the base value, finishing the inspection, and recording the concentration threshold value of the nasal cavity excitation reagent; otherwise, the result is negative, and the inspection A is required;

and (5) checking C: spraying 1 spray to the nasal cavity at both sides of a patient by using a nasal cavity stimulating spray containing a nasal cavity stimulating reagent stock solution, filling a VAS score of rhinitis symptoms immediately after 10 minutes, judging that a stimulating test result is positive if the VAS score is increased by more than or equal to 55 points relative to a basic value, and recording a nasal cavity stimulating reagent concentration threshold; otherwise, judging the result of the patient arousal test as negative.

The embodiment of the invention also provides application of the house dust mite allergen pricking liquid in a nasal cavity allergen excitation test, wherein the house dust mite allergen pricking liquid is derived from a mite allergen skin pricking kit, the mite allergen skin pricking kit is named as Anci in Chinese, Soluprick SQ in English, and ALK-Abello A/S is a manufacturer. By applying the house dust mite allergen pricking liquid to the kit, the application of the house dust mite allergen pricking liquid is widened, and meanwhile, a standard reagent which is safe, economical, simple to operate, high in repeatability, high in patient acceptance and easy to popularize is provided for a nasal cavity excitation test by a spraying method. The present invention is further illustrated by the following examples.

EXAMPLE-preparation of nasal Cavity stimulating kit

This example illustrates a nasal cavity stimulation kit with the specification of 2mL nasal cavity stimulation spray and vehicle spray per bottle.

First, the material source of the nasal cavity stimulating kit

1. Stock solution of nasal cavity stimulating reagent

The nasal cavity stimulating reagent stock solution used in this example was house dust mite allergen prick solution (specification: 2 mL/bottle) in a mite allergen skin prick kit, and the manufacturer was ALK-Abello A/S. It should be further noted that, the mite allergen skin prick kit has a trade name (Chinese) of Anci, a trade name (English) of Soluprick SQ, a dosage form (Chinese) of pricking liquid, a specification (Chinese) of 2 mL/bottle, a packaging specification (Chinese) of 4 bottles/boxes, and 4 bottles of reagents of 2 mL/bottle, 2 mL/bottle and 2 mL/bottle of house dust mite allergen prick liquid, positive control prick liquid and negative control prick liquid, respectively. The nasal cavity exciting reagent stock solution used in the invention is house dust mite allergen prick liquid. The house dust mite allergen pricking liquid consists of a house dust mite allergen and a solvent, wherein the concentration of the house dust mite allergen in the house dust mite allergen pricking liquid is 10HEP (HEP is a biological activity unit of allergen concentration), and the solvent is prepared from phenol, sodium dihydrogen phosphate, disodium phosphate, sodium oxide, glycerol and water for injection.

2. Solvent

The solvent in the embodiment of the invention is the solvent in the stock solution of the nasal cavity stimulating reagent, namely the solvent in the dermatophagoides pteronyssinus allergen pricking liquid, and the manufacturer of the solvent is ALK-Abello A/S.

The above-mentioned information on the suppliers of the nasal cavity stimulating reagent stock solution and the vehicle is merely to better describe the range of the ratio and the effect of the examples, and the kinds and suppliers of the nasal cavity stimulating reagent stock solution and the vehicle of the present invention are not limited thereto.

3. Spray bottle

The spray bottle is aseptic, brown light-resistant, push type spray bottle, and the capacity is 5mL, be provided with medicine spraying agent pump on the spray bottle, each spouts even aerial fog, and the spray volume of once spouting when pressing is 0.05mL, the nozzle of spray bottle is detachable construction, can replace, during the use, can prepare a plurality of disposable nozzles to make things convenient for many people to use, the spray bottle manufacture factory that this example used is the Apta company. The above information on the supplier of the spray bottle is merely to better describe the effect of the example, and the supplier of the spray bottle used in the kit of the present invention is not limited thereto.

Composition of nasal cavity excitation kit

This example nasal cavity elicitation kit includes:

2 mL/bottle of solvent spray;

1/1000 concentration of exciting reagent stock solution spray 2 mL/bottle;

1/100 concentration of exciting reagent stock solution spray 2 mL/bottle;

1/10 concentration of exciting reagent stock solution spray 2 mL/bottle;

2 mL/bottle of the excitation reagent stock solution spray.

Preparation of nasal cavity excitation kit

The preparation method of the nasal cavity excitation kit of the embodiment is as follows:

(1) preparing a solvent spray: transfer 2000. mu.L of solvent into spray bottle with pipette, bottle label "solvent".

(2) Preparing 1/1000 concentration excitation reagent stock solution spray: firstly, transferring 1998 mu L of solvent into a spray bottle by using a micropipette, then sucking 2 mu L of allergen solution into the spray bottle by using the micropipette, and blowing and beating the allergen solution uniformly, wherein the label of the bottle label is '1/1000 concentration excitation reagent stock solution';

(3) preparing 1/10 concentration stimulating agent stock solution spray: firstly, transferring 1980 mu L of solvent into a spray bottle by using a liquid transfer gun, then sucking 20 mu L of raw dermatophagoides pteronyssinus solution into the spray bottle by using a micropipettor, and blowing and stirring uniformly, wherein the label is '1/100 concentration excitation reagent raw solution';

(4) preparing 1/10 concentration stimulating agent stock solution spray: transferring 1800 mu L of solvent into a spray bottle by using a liquid transfer gun, sucking 200 mu L of raw dermatophagoides pteronyssinus solution into the spray bottle by using a micropipettor, blowing and uniformly mixing, wherein the label of the bottle label is '1/10 concentration exciting reagent raw solution'.

(5) Preparing an exciting reagent stock solution spray: 2000. mu.L of the house dust mite stock solution was transferred into a spray bottle using a pipette, and the bottle label was "challenge reagent stock solution".

After the preparation is finished, the spray is stored at the temperature of-4 ℃, and is placed to the room temperature at the room temperature of 26 ℃ before use.

The advantages of the nasal cavity stimulation kit of the present invention are further illustrated by the efficacy tests performed on the final product of example one of the present invention.

Effects of the embodiment

1. The kit is used for selecting objects, and 119 volunteer patients with high symptom conformity to the allergic rhinitis are detected by the kit.

1) Indications of

(1) When the medical history does not match with the SPT and/or blood sIgE result or the diagnosis is difficult, NAPT is used for judging;

(2) used for diagnosing Allergic Rhinitis (AR), especially for patients with positive results on various allergens and confirming the clinical relevance of specific allergens;

(3) determining the extent of nasal cavity reaction to a particular allergen prior to allergen-specific immunotherapy;

(4) for evaluating the effectiveness of a rhinitis treatment drug;

(5) for the etiology of occupational allergenic respiratory diseases;

(6) NAPT is studied in assessing the severity of nasal symptoms, sensitivity to allergens, and the pathophysiological mechanisms of the nose triggered by allergen stimuli in patients.

2) Contraindication

(1) The patient is in the elution phase of any drug that may affect the nasal mucosal response. Such as oral antihistamines: 48 hours to 1-2 weeks; nasal antihistamines: 4-5 days; nasal hormones: 48-72 hours; oral administration of hormones: 2-3 weeks; sodium cromogroside: 1-3 weeks; nasal decongestant: 2 days; tricyclic antidepressants: 2-3 weeks; non-steroidal anti-inflammatory drugs: 1 week; reserpine and clonidine hypotensor: 3 weeks;

(2) the patient is in the rhinitis attack stage and has obvious nasal symptoms of nasal obstruction, watery nasal discharge, sneeze and the like; acute bacterial or viral rhino-sinusitis episode;

(3) nasal septal deflection, nasal polyps and other nasal space occupying lesions with severe nasal obstruction symptoms;

(4) the prior history of severe anaphylactic reaction;

(5) patients with uncontrolled asthma or severe chronic obstructive pulmonary disease or cardiopulmonary disease with epinephrine;

(6) other serious systemic diseases, such as malignant tumor, autoimmune disease;

(7) a gestational period;

(8) children under 5 years old (relative contraindication).

2. Procedure for the preparation of the

SPT and sIgE examination is performed with reference to guidelines for allergic Rhinitis and its effect on Asthma (Bousquet J, Helling PW, Agache I, et al, Allergy Rhinitis and its Impact on Asthma (ARIA) Phase 4(2018), Change management in allergic Rhinitis and astma multicombining using mobile technology J Allergy Clin Immunol,2019,143(3):864-879.) and literature (Wanghua, Malin, Wangchong Shuo, et al. consensus on allergen skin prick test [ J ] Beijing medicine 2020,42(10):966-985.DOI: 10.15932/j.0253-9713.2020.10.912.).

1)SPT

(1) Preparing an article: points out needles. ② allergen pricking liquid. ③ positive contrast solution: histamine. Fourthly, negative contrast liquid: a solvent. 75% medical alcohol and dry cotton stick. Sixthly, the special measuring scale for SPT. The special report sheet for SPT. The marking pen and the transparent adhesive tape.

(2) The operation method comprises the following steps: arranging the prepared articles on a special tray in sequence. Selecting proper prick parts. ③ 75 percent alcohol is used for disinfecting the patient and pricking the test area. Fourthly, the marking pen is used for marking the prick test area, the interval between adjacent positions is at least 2cm, and cross contamination is avoided. Dropping allergen pricking liquid, positive contrast liquid and negative contrast liquid to the corresponding mark of pricking test area to avoid cross contamination. Sixthly, tightening the skin and avoiding blood vessels, vertically puncturing the skin through the medicine drops at an angle of 90 degrees by the pricking needle (pricking the skin, so that a small amount of pricking liquid below the needle point enters the skin, and lifting the pricking needle after the pricking needle stays for 1 s). Seventhly, pressing the skin lightly by using a cotton stick, and sucking away prick liquid; if the patient has strong reaction, the prick liquid must be wiped off immediately; for those who do not have strong reaction, the prick liquid wiping can be completed 5-10 min after the prick is completed or before the result is interpreted.

2) sIgE detection

Immediately after drawing 5mL of peripheral venous blood from the patient following a strict aseptic procedure, the serum was separated and collected using a centrifuge (Thermo ST 16R; America), and the concentration of serum sIgE was measured using the UNICAP system (Pharmacia, Uppsala, Sweden).

3)NAPT

The nasal allergen challenge test comprises three parts of basal assessment before challenge, challenge with vehicle and challenge with allergen, and rhinitis symptoms VAS scoring is required to be carried out immediately after 10 minutes of each challenge.

VAS score specification: this is easily understood by the guidelines for allergic Rhinitis and its effect on Asthma (Bousquet J, Helling PW, age I, et al, organic rhinonitis and its Impact on Asthma (ARIA) Phase 4(2018): Change management in alloc Rhinitis and activity multiple using mobile Clin Immunol,2019,143(3):864-879.), Visual Analogue Score (VAS) and AR symptom severity (ref: Bousquet PJ, patent C, NeukirF, et al, Visual Analogue Scale can be caned) and the severity of AR symptoms (refer to Bousquet J, clinical C, Neukir F, et al, Visual Analogue Scale can be used) in patients with Asthma (refer to Bousquet J, age I, and about 4. ARIA), and this is easily understood by the guidelines for Asthma (ARIA, ARE 4. this method is easily understood by the guidelines for patients.

Fig. 3 is a VAS sample table in an example of the present invention. Referring to FIG. 3, a straight line of 100mm length is shown, with the left most point being 0, which represents no symptoms, and the right most point being 100, which represents severe symptoms, and the heavier the symptoms, the higher the score. The patients give scores for their symptoms of nasal obstruction, watery nasal discharge, sneezing and rhinocnesmus ranging from 0-100 mm (no symptoms to extremely severe symptoms). When the sum of the VAS values of the four symptoms is more than or equal to 55 points relative to the change of the basic value, the nasal cavity stimulation test result is judged to be positive, and a corresponding positive threshold (concentration of the stimulation liquid) is recorded.

Fig. 4 is a flow chart of the application of the nasal cavity excitation kit in the nasal cavity allergen excitation test according to the embodiment of the invention. Referring to fig. 4, the first step is to fill in the basic rhinitis symptoms VAS score. The patient had to sit still in the examination room (temperature 26 ℃, humidity 40% -60%) for 30 minutes before applying no agent to the nasal cavity, and then filled out the basic rhinitis symptoms VAS assessment and scored as the control basis.

And the second step, carrying out a solvent excitation test. Spraying 1 spray of the "solvent spray" into nasal cavity of both sides of the patient, and filling up VAS score of rhinitis symptoms immediately after 10 minutes to remove nasal non-specific hyperreactivity. When the VAS score is increased by less than 55 points relative to the basic value, the interference of the solvent to the test can be eliminated, the result is negative, and the next detection (namely, the third step) can be carried out; otherwise, nasal non-specific hyperreactivity, indicating a "positive" result, requires discontinuing further examination (i.e., the third step) and re-examination of the patient two weeks later.

And thirdly, carrying out an allergen excitation test. Spraying 1 spray of '1/1000 concentration stimulating reagent stock solution spray' into nasal cavities at both sides of a patient respectively, filling in a VAS score of rhinitis symptoms immediately after 10 minutes, if the VAS score is increased by more than or equal to 55 points relative to a basic value, judging that the stimulating test result is 'positive', ending the examination, and recording the concentration threshold of the stimulating solution; otherwise "negative", test A is required.

Checking A: spraying 1 spray of '1/100 concentration stimulating reagent stock solution spray' into nasal cavities at both sides of a patient respectively, filling in a VAS score of rhinitis symptoms immediately after 10 minutes, if the VAS score is increased by more than or equal to 55 points relative to a basic value, judging that the stimulating test result is 'positive', ending the examination, and recording the concentration threshold of the stimulating solution; otherwise "negative", it is necessary to perform a check B.

And B, checking: spraying 1 spray of '1/10 concentration stimulating reagent stock solution spray' into nasal cavities at both sides of a patient respectively, filling in a VAS score of rhinitis symptoms immediately after 10 minutes, if the VAS score is increased by more than or equal to 55 points relative to a basic value, judging that the stimulating test result is 'positive', ending the examination, and recording the concentration threshold of the stimulating solution; otherwise "negative", it is necessary to perform a check C.

And (5) checking C: spraying 1 spray to the nasal cavities at two sides of a patient by using an 'exciting reagent stock solution spray', filling a VAS score of rhinitis symptoms immediately after 10 minutes, judging that an exciting test result is positive if the VAS score is increased by more than or equal to 55 points relative to a basic value, and recording an exciting solution concentration threshold; otherwise, judging the result of the patient arousal test as negative.

3. Analysis of results

The detection of SPT and sIgE is a commonly used allergen detection method at present, and the SPT and sIgE detection have certain consistency, the SPT detection result and the serum sIgE level have good positive correlation, and the SPT detection result and the serum sIgE level are mutually complementary but not replaceable (reference: de Vos G. skin testing serum server-specific IgE testing: white is for differentiating the allergic aeroallergene sensing and predicting clinical allergery [ J ]. Curr Allergy elevation Rep,2014,14: 430.). Therefore, the accuracy of the detection result of the kit can be verified according to the detection of SPT and sIgE.

As shown in table 1, the results of the examination of the examples of the present application using the combination of two examination methods of house dust mite allergens SPT and sIgE show that: in 119 patients with clinical symptoms and signs suspected of house dust mite allergic rhinitis, 69 patients were diagnosed with AR and 50 patients with NAR by examination of house dust mite allergens SPT and sgle. In addition, 69 patients diagnosed with AR by symptoms, house dust mite SPT and sIgE were positive in NAPT results after examination by the house dust mite allergen spray nasal cavity excitation test and further confirmed to be AR. NAPT results were negative in 5 patients, and these 5 patients were defined as "sensitized" status rather than AR. If this fraction of "sensitized" patients (approximately 7.2% of AR) were treated with house dust mite AIT, this class of patients would be expected to have poor specific immunotherapy and would be very difficult to benefit from AIT. The results show that the detection results of the nasal cavity excitation kit for detecting Allergic Rhinitis (AR) and SPT and sIgE have high consistency, and the NAPT further defines the allergen for AR patients before AIT, so that the method is of great significance. Patients with poor AIT treatment, who are indistinguishable from SPT and sIgE, can be further distinguished by NAPT using the kit of the present invention, and are of reference value in helping to determine the type of allergic rhinitis and whether AIT treatment is available or predicted.

TABLE 1 SPT, sIgE, NAPT assay results

In addition, in 50 patients who were diagnosed with NAR by symptoms, dermatophagoides pteronyssinus SPT and sIgE, after examination by the dermatophagoides pteronyssinus allergen nasal cavity excitation test, NAPT results were positive in 18 (36%) of the patients, and they were further diagnosed with LAR. Without NAPT, a percentage of 36% of LAR patients will not be diagnosed from NAR, missing a diagnosis, missing the opportunity for AIT treatment. The results further show that for the patients diagnosed with dust mite SPT and serum sIgE as NAR, whether the disease is LAR can be further diagnosed by using the nasal cavity excitation kit for detection, so that reference is provided for determining a further treatment scheme, and the diagnosis accuracy of the patients with house dust mite allergic rhinitis is improved.

In conclusion, the nasal cavity excitation kit has obvious advantages in the aspects of diagnosis and detection of allergic rhinitis. For AR patients detected using conventional means such as SPT and/or serum sIgE, the allergens can be further defined for these AR patients prior to AIT, and the efficacy of the AR patients detected by SPT and/or serum sIgE for AIT treatment can be further assessed, providing reference for the formulation of clinical treatment regimens.

In addition, NAR patients detected by conventional means such as SPT and/or serum sIgE can be further diagnosed as LAR by using the nasal cavity excitation kit, so that reference is provided for clinically determining further treatment schemes, and the diagnosis accuracy of allergic rhinitis patients is improved.

Although the present invention is disclosed above, the present invention is not limited thereto. Various changes and modifications may be effected therein by one skilled in the art without departing from the spirit and scope of the invention as defined in the appended claims.

Claims

1. A nasal cavity excitation kit, comprising:

a solvent spray, which contains a solvent.

2. The nasal cavity stimulation kit according to claim 1, wherein the nasal cavity stimulation spray is capable of spraying the nasal cavity stimulation reagent uniformly and quantitatively, and the solvent spray is capable of spraying the solvent uniformly and quantitatively.

3. The nasal cavity stimulating kit according to claim 1 wherein the nasal cavity stimulating spray contains only a single allergen per unit size, said allergen being an inhalant allergen.

4. The nasal cavity challenge kit of claim 1 wherein the allergens include, but are not limited to, dermatophagoides farinae allergen, dermatophagoides pteronyssinus allergen, ragweed allergen, alternaria allergen, dog hair allergen, cat hair allergen, phoenix tree allergen, birch allergen, grass pollen allergen, ragweed pollen allergen, tropical mite allergen, blattella germanica allergen, alternaria allergen, aspergillus niger allergen, artemisia argyi allergen, phoenix tree allergen, willow allergen, poplar allergen, birch allergen, hop allergen, elm pollen allergen, or cedar pollen allergen.

5. The nasal cavity excitation kit according to claim 1, wherein the solvent comprises one or more of phenol, sodium dihydrogen phosphate, disodium phosphate, sodium oxide, glycerin, and water for injection.

6. The nasal cavity stimulation kit according to claim 1, wherein the nasal cavity stimulation reagent and the solvent are respectively contained in a spray bottle, a drug spray pump is arranged on the spray bottle, and each spray of the nasal cavity stimulation spray and the solvent spray is 0.04mL to 0.06 mL.

7. The nasal cavity excitation kit according to claim 1, wherein the nasal cavity excitation reagent comprises a gradient concentration of a stock solution of the nasal cavity excitation reagent, and the stock solution of the nasal cavity excitation reagent is dermatophagoides pteronyssinus allergen prick liquid.

8. The nasal cavity challenge kit of claim 7 wherein the concentration of allergen in the nasal cavity challenge reagent stock solution is from 8HEP to 10 HEP.

9. The nasal cavity excitation kit of claim 7, wherein the gradient concentration of the nasal cavity excitation reagent stock solution comprises: 1/1000 nasal cavity stimulating agent stock solution, 1/100 nasal cavity stimulating agent stock solution, 1/10 nasal cavity stimulating agent stock solution and nasal cavity stimulating agent stock solution.

10. A method of preparing a nasal cavity stimulation kit for use in preparing a nasal cavity stimulation kit as claimed in any one of claims 1 to 9, the method comprising:

preparing a solvent;

preparing a nasal cavity stimulating reagent;

11. The method of making a nasal cavity stimulation kit of claim 10, wherein the method comprises:

12. Use of a nasal cavity challenge kit in a nasal cavity allergen challenge test, wherein the nasal cavity challenge kit is according to any one of claims 1 to 9.

13. The use of a nasal cavity challenge kit according to claim 12 in a nasal cavity allergen challenge test comprising:

14. Use of the nasal cavity challenge kit of claim 13 in a nasal cavity allergen challenge test wherein the reference value is set to 55 points.

15. The use of a nasal cavity challenge kit according to claim 12 in a nasal cavity allergen challenge test comprising:

16. The application of the house dust mite allergen pricking liquid in a nasal cavity allergen excitation test is characterized in that the house dust mite allergen pricking liquid is derived from a mite allergen skin pricking kit, the mite allergen skin pricking kit is named as Anci in Chinese, and is named as Soluprick SQ in English, and the manufacturer is ALK-Abello A/S.