CN102533933A - Method, reagent and kit for detecting oral inflammatory index, and applications thereof - Google Patents
Method, reagent and kit for detecting oral inflammatory index, and applications thereof Download PDFInfo
- Publication number
- CN102533933A CN102533933A CN201010588508XA CN201010588508A CN102533933A CN 102533933 A CN102533933 A CN 102533933A CN 201010588508X A CN201010588508X A CN 201010588508XA CN 201010588508 A CN201010588508 A CN 201010588508A CN 102533933 A CN102533933 A CN 102533933A
- Authority
- CN
- China
- Prior art keywords
- solution
- oral
- mammals
- neutrophils
- cell
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Images
Abstract
The invention provides a solution for releasing neutrophile leucocyte cells in oral cavity, and a kit comprising the solution. In addition, the invention also provides a method for detecting oral inflammatory index of mammals. The oral inflammatory index can be expressed by the count of the neutrophile leucocyte cells or the concentrations of the specific components or products of the neutrophile leucocyte cells obtained from the oral cavity. Furthermore, the invention also discloses a method for assistant detection of oral inflammation treating condition of the mammals.
Description
Technical field
The invention belongs to the medical examination field, particularly, the present invention relates to detect oral inflammation exponential method, related reagent and test kit and the application thereof of this method.
Background technology
Inflammation is a kind of important immunoreation of tackling infection, damage or various harmful situations.Inflammatory process is very important to body, damages, kills and wounds in the process of pathogenic micro-organism and tumour because it serves to repair.Yet persistent inflammation is deleterious, because inflammatory cell discharges the enzyme of toxic molecule and infringement normal cell and tissue.
Non-solution property inflammation is a kind of main drive of disease; It is promoting the pathological process of multiple disease significantly, and said disease comprises: atherosclerosis, mellitus, obesity, cancer, chronic obstructive pulmonary disease, asthma, inflammatory bowel disease, nerve degenerative diseases, multiple sclerosis or rheumatoid arthritis morbidity.
Detecting inflammatory process is very important concerning the doctor, because its appearance means the existence of major disease, injury or harmful situation.The disease example that often has serious inflammation characteristic has: myocardial infarction, active tuberculosis, osteomyelitis (infection of bone), rheumatoid arthritis, cholecytistis (infection courage) and pyelonephritis (kidney infection) and cancer diffusion.
The patient who suffers from obvious inflammation tends to show well-known inflammation sign and symptom, like fever, weak and poor appetite.Yet these sings and symptomses be both insensitive, also do not show the existence of inflammation specifically.Before some disease and physical appearance can be more specifically, diagnosable illness occurs, inflammatory reaction appearred.Therefore, detect the appearance of inflammatory symptom, will help diagnosing more in time and treating these diseases and discomfort.
It is erythrocyte sedimentation rate or ESR that famous and the most widely used at present blood inflammation detects index.ESR is found that by Fahraeus Wintrobe and Westergren improve and promote, and formed Wei Shi blood sedimentation rate or WESR thoughts on the overall rising of inflammatory protein, measure red corpuscle settled distance in 60 minutes.Doctors attempted once to develop that other are more easily or faster, or more responsive or concrete inflammation detection method.These detections comprise c reactive protein or CRP, white blood cell count(WBC), orosomucoid albumen, red corpuscle or oxyphorase and Fibrinogen (with reference to U.S. Patent number 5137832, U.S. Patent number 4843869, U.S. Patent number 5506145).
Yet for the oral cavity, the oral cavity is the environment that comes with blood separating, and the level of inflammation in the blood can not be reflected in the situation of oral inflammation.Therefore, present blood inflammation detection method can not be used to the diagnosis at oral inflammation.
The oral cavity is to get into the intravital passage of people, and many incidents take place at this.Food is accepted in the oral cavity, and the solid food particles is diminished, and mixes with saliva, starts digestive process.Oral mucosa and saliva glandular secretion provide lubricating fluid, help verbal exposition, swallow and food digestion.Therefore, the oral cavity infect especially easily, damage or various harmful situation.In addition, distinctive intraoral disease and the discomfort of occurring in of a cover arranged in the oral cavity, comprise carious tooth, periodontal, stomatocace etc.Some situation such as bad sense of smell, halitosis even snoring also maybe be relevant with oral Health.Inflammatory reaction by due to the different reasons possibly appear in the mouth simultaneously.The inflammatory factor of different sources possibly influence each other between the different event, because may form an interactional level in a local relatively environment of sharing.This should be different from body inner blood environment.
Therefore, press for the method for detection oral inflammation level or the method and the related prods thereof of auxiliary detection oral inflammation level at present.
Summary of the invention
In order to solve the problems of the technologies described above, the present invention provides aided detection method and the related prods that relates to this field.
For helping to understand the present invention, some terms have been defined below.The term of this paper definition has the implication of those of ordinary skill in the related art's common sense of the present invention.
Unless otherwise indicated, the quantizating index of term used herein " oral inflammation index " the inflammation degree that is meant in the oral cavity to be taken place.In this application; Said " oral inflammation index " specifically by the representative of level relatively of the amount of the neutrophils cell that can collect in the oral cavity, its existence that can be used for the said Mammals oral inflammation of auxiliary judgment whether, severity and/or improvement situation.
Unless otherwise indicated, term used herein " personalized oral inflammation index " representes that by the growth multiple of neutrophils cell count in the detected object oral cavity or its specific component (or product) concentration this exponential calculation formula is following:
I
i＝n
t/n
h
I wherein
iOral inflammation index for personalization; n
tBe neutrophils cell number or its specific component (or product) concentration of from detected object, obtaining; n
hNeutrophils cell number or its specific component (or product) concentration under the health condition of NIP, obtained for this detected object.
Unless otherwise indicated, term used herein " comparative oral inflammation index " is by neutrophils cell count or its specific component (or product) concentration of detected object, with respect to the health of the NIP growth multiple with reference to the person.This exponential calculation formula is following:
I
c＝n
t/n
r
I wherein
cBe comparative oral inflammation index; n
tBe neutrophils cell number or its specific component (or product) concentration of from detected object, obtaining; n
rBe the health of NIP neutrophils cell number or its specific component (or product) concentration with reference to the person.According to an embodiment of the present invention, the health of NIP is with reference to person's neutrophils cell number (n
r) be set at 10,000.
In order to solve the problems of the technologies described above, an object of the present invention is to provide a kind of solution that is used to detect neutrophils cell in oral inflammation index and/or the delivering oral.
Another object of the present invention provides the test kit that is used to detect the oral inflammation index and/or detects neutrophils cell quantity in the oral cavity.
Another object of the present invention is above-mentioned solution and the purposes of test kit in preparing the product that detects neutrophils cell quantity in the oral cavity.
Further object of the present invention provides the method that detects oral cavity inflammation index in the Mammals oral cavity.
A the present invention also purpose provides the method that a kind of auxiliary detection Mammals oral inflammation is treated situation.
The objective of the invention is to realize through following technical scheme.
On the one hand, the present invention provides a kind of solution that is used for neutrophils cell in the delivering oral, it is characterized in that, said solution comprises salt ion and the acid ion that is used for neutrophils cell in the delivering oral;
Preferably, said salt ion and acid ion are provided by compound, and said compound is selected from Repone K, one or more of sodium-chlor and sodium phosphate;
Said solution can further comprise the compound that is used to provide cell institute energy requirement; Preferably, provide the compound of said cell institute energy requirement to be: D-glucose.
Said solution can further comprise and be used to regulate the pH value of this solution and the compound of shock-absorbing capacity; Preferably, said compound is selected from anhydrous potassium dihydrogenphosphate, one or more of sodium phosphate and anhydrous phosphoric acid hydrogen two ammoniums.
In addition, said solution also can further comprise and be used for painted compound; Preferably, said compound is: phenol red sodium.
Further preferably, comprise in the said solution: 0.01-10.00 gram/L Repone K, 0.01-10.00 gram/L anhydrous potassium dihydrogenphosphate; 0.01-10.00 gram/L sodium-chlor;, the anhydrous binary sodium phosphate of 0.01-10.00 gram/L, 0.01-10.00 gram/LD-glucose and the phenol red sodium of 0.01-0.02 gram/L;
More preferably, said solution comprises 0.4 gram/L Repone K, 0.06 gram/L anhydrous potassium dihydrogenphosphate, 8.0 gram/L sodium-chlor, the anhydrous binary sodium phosphate of 0.04788 gram/L, 1.0 gram/L D-glucose and the phenol red sodium of 0.011 gram/L.
And said solution can be the aqueous solution, spirituous solution or glucose solution;
Preferably, the ethanol concn in the said spirituous solution is 1-70% (V/V); Or
Preferably, the glucose concn in the said glucose solution is 0.1-15% (g/ml).
On the other hand, the present invention provides a kind of test kit that is used to detect neutrophils cell quantity in the oral cavity, and said test kit comprises the above-mentioned solution that is used for neutrophils cell in the delivering oral.
In the mentioned reagent box, can further include fixing agent that is used for the fixing said neutrophils cell of specificity and/or the staining agent that is used for said neutrophils cell is carried out specific stain;
Preferably, said fixing agent is a formaldehyde, and said staining agent is a SP 15 Lemon Yellow.
In addition, said test kit can further include special composition or the required detection reagent of product that detects the neutrophils cell; Preferentially, said specific component is: neutrophil elastase, cathepsin G, myeloperoxidase, one or more of Lf lactoferrin and gelatinase; Its detection reagent comprises required antibody and the relevant detection reagent thereof of this differential protein of detection; Its detection reagent is the required reagent of integrated enzyme reaction.
Another aspect, the present invention provides above-mentioned solution or the purposes of mentioned reagent box in preparing the product that detects neutrophils cell quantity in the Mammals oral cavity; Preferably, said Mammals is the people.
On the one hand, the present invention provides a kind of detection Mammals oral inflammation exponential method again, and said method comprises: the quantity that detects neutrophils cell in the sample that comprises said Mammals oral cavity inclusion;
Preferably, said oral inflammation index is personalized oral inflammation index or comparative oral inflammation index;
Preferably, said Mammals is the people.
Wherein, aforesaid method preferably specifically comprises:
1) said Mammals is gargled with an amount of solution, thereby obtains to comprise the solution of said Mammals oral cavity inclusion;
2) use the fixing neutrophils cell in the solution that step 1) obtains of fixing agent;
3) using the staining agent will be through step 2) fixed neutrophils cell dyes and counts;
Preferably, in step 2) preceding, the centrifugal said solution that comprises Mammals oral cavity inclusion of elder generation; Further preferably, said centrifugal speed is 20-100g, and the time is 10-30 minute; More preferably, said centrifugal speed is 20g, and the time is 10 minutes.
And the solution in the said step 1) is preferably the above-mentioned solution that is used in the Mammals oral cavity, discharging the neutrophils cell;
Said step 2) fixing agent in is preferably formaldehyde; Further preferably, said fixing agent is the formalin of 2.5% (g/ml);
Staining agent in the said step 3) is preferably SP 15 Lemon Yellow; Further preferably, said staining agent is the SP 15 Lemon Yellow aqueous solution of 1.6% (g/ml).
In addition; Said method further comprises through the quantity of detected neutrophils cell in the said sample and the quantity of Mammals to be detected intraoral neutrophils cell under state of health are compared, calculates the oral inflammation index of said mammiferous personalization.
Alternatively; Said method further comprises through the quantity of detected neutrophils cell in the said sample and the Mammals under the state of health quantity with reference to the intraoral neutrophils cell of person is compared, calculates said mammiferous comparative oral inflammation index; Preferably, the Mammals under the said state of health is 10000 with reference to the quantity of the intraoral neutrophils cell of person.
The present invention also provides other a kind of detection Mammals oral inflammation exponential method; Said method comprises: the content that detects neutrophils cell-specific composition in the sample comprise said Mammals oral cavity inclusion or product; Said special composition such as neutrophil elastase; Cathepsin G, myeloperoxidase, Lf lactoferrin and gelatinase.These differential proteins can adopt this area routine techniques, use corresponding antibody, detect through integrated enzyme reaction.；
Preferably, said oral inflammation index is personalized oral inflammation index or comparative oral inflammation index;
Preferably, said Mammals is the people.
In addition; Said method further comprises through the content of detected neutrophils cell-specific composition or product in the said sample and the Mammals to be detected content of intraoral neutrophils cell-specific composition or product under state of health is compared, calculates the oral inflammation index of said mammiferous personalization.
Alternatively; Said method further comprises through the content of detected neutrophils cell-specific composition or product in the said sample and the Mammals under the state of health content with reference to person's intraoral neutrophils cell-specific composition or product is compared, calculates said mammiferous comparative oral inflammation index.
Also on the one hand, the present invention provides a kind of method of auxiliary detection Mammals oral inflammation treatment situation, and said method comprises:
Before the Mammals oral inflammation is treated,, detect the intraoral oral inflammation index of Mammals respectively, relatively treat before then and oral inflammation index afterwards according to above-mentioned each method with afterwards;
Preferably, said oral inflammation index is personalized oral inflammation index or comparative oral inflammation index;
Preferably, said Mammals is the people.
Below be detailed description of the present invention:
At first, the present invention has disclosed the generation of former the unknown and has embedded the inflammation source in the oral cavity mucous membrane tissue through following research.Remove this inflammation source, chronic periodontitis that can heal the sick and regular ulcer.This shows, the inflammation that embeds mucous membrane tissue is a kind of inducement of the oral disease relevant with inflammation.On the basis of this research, the present invention has set up the method for auxiliary diagnosis oral inflammation through detecting oral cavity inflammation index in the Mammals oral cavity.
The patient of selected this research of participation 45 years old suffers from the periodontitis in 15 years, confirms to have this disease by three different periodontitis expert diagnosis.This patient's every three months carries out the specialty cleaning with regard to needs to the periodontitis training.This patient also often suffers the puzzlement (Figure 1A) of stomatocace.
The special cleaning means of a Y shape of employing thoroughly cleans this patient's oral cavity, comprises cleaning mucous membrane and gingiva tissue.The special of this instrument has been optimized cleansing power, also can have been avoided injuring any related organization (Figure 1B), is referred to as to remove scorching brush (specifically referring to Chinese invention patent application CN201010109442.1).Oral cavity mucous membrane tissue is brushed wiping, water flushing then with removing scorching brush.The residue that turns white discharges from oral cavity mucous membrane tissue, has the appearance of bloodstain once in a while, but the continuous bleeding phenomenon does not take place.
Subsequently, ulcer sampling point (ulcer-like spot) occurs on the mucous membrane tissue of nigh mucous membrane gum intersection (Fig. 1 C).Surprisingly, brown secretory product discharges (Fig. 2 A) from the ulcer sampling point.This secretory product dyes with T ü rk ' s solution, under fluorescent microscope, observes then.Obviously, contain a large amount of neutrophils (neutrophil) (Fig. 2 B) in this secretory product, these cells all contain typical polymorphic nucleus characteristic (Fig. 2 C).This sample is dyeed by SP 15 Lemon Yellow (Acridine orange), under fluorescent microscope, is proved to contain neutrophils (Fig. 2 D).
As if this brown secretory product contains a neutrophils with structural unit and assembles colony, and the neutrophils colony that these pool together is referred to as stomatitis spot (oral inflammatory plague).The neutrophils that in brown secretory product, occurs is a kind of inflammatory symptom; Neutrophils discharges can damage tooth and the lytic enzyme of tissue.From a ulcer sampling point, can discharge more than 10 stomatitis spots, the ulcer sampling point is cured subsequently.The stomatitis spot will can not discharge from the position of suffering from the ulcer sampling point more then.The stomatitis spot is present in the time marquis in the ulcer sampling point, and the patient can have a kind of temperature and uncomfortable feeling at this position.In case the stomatitis spot is released out, the patient just feels a kind of sensation as if relieved of a heavy load at once.These show that all the stomatitis spot is a kind of inflammatory reaction.
By inference, the stomatitis spot is embedded in (Fig. 3) in the mucous membrane tissue.Support the fact of this idea to be, need take time discharges the stomatitis spot with strength from mucous membrane tissue.A lot of stomatitis spots can pool together, because mucous membrane tissue is folding, the stomatitis spot is embedded into wherein, forms a kind of " knot " columnar structure, is referred to as " stomatitis knot " (oral inflammatory knotitis).The stomatitis knot can cause the visible inflammatory symptom, like above-mentioned said ulcer sampling point (Fig. 3).Visible white residue or throw out possibly caused by the necrocytosis due to the neutrophils at ulcer sampling point place.
Through removing the process that scorching brush cleans, find to contain in this patient oral cavity the ulcer sampling point more than 12.These ulcer sampling points are present near the mucous membrane gum intersection, tongue is following, near the palatine tonsil, and inner at cheek; These all are the places that can not be cleared up by current oral cavity cleaning measure easily.Some ulcer sampling point possibly be because due to its place position, size and complexity than other more difficult removing.The number of the stomatitis spot that from a ulcer sampling point, discharges is different, and what have has about 10, have up to more than 100.The stomatitis spot that has is a brown, and the stomatitis spot that has is for yellow.The volume of stomatitis spot also changes to some extent, have about 10 microlitres, have up to more than 200 microlitres.The stomatitis spot total amount that from this patient, discharges is estimated more than 10 milliliters.The estimation of the concentration of neutrophils is 10 in the stomatitis spot
7About/ml.Therefore, contain at least 108 neutrophils in this patient's the oral cavity mucous membrane tissue.This possibly be an inflammatory reaction with remarkably influenced power, causes oral disease therefrom.
After 3 to 5 in 12 ulcer sampling points were eliminated, this patient noticed that he has not had periodontal disease.This shows removes the effective measure that the ulcer sampling point is the treatment periodontitis.Amazingly be, along with the further removing of ulcer sampling point, the also less generation of patient's stomatocace.After the ulcer sampling point was removed end, this patient had also no longer suffered the puzzlement of stomatocace.This shows that oral inflammation is the inducement that causes the oral cavity recurrent ulcer.
This patient notices that also after the ulcer sampling point was removed, his problem of bad breath also was able to eliminate.Can imagine that the mikrobe that is trapped in ulcer sampling point the inside is the reason (Fig. 3) that causes halitosis.What is interesting is that his sensory system has improved, he can smell more smell.By inference, the neutrophils in the oral cavity may be destroyed the sensory cell in the respiratory system.Therefore, remove the ulcer sampling point and can treat gum disease, also be of value to healthy.
These researchs show, the deficiency of current oral sanitary habit has caused food residue (Fig. 2 B), the accumulation of cell debris (white depositions is like Fig. 1) and mikrobe (problem of bad breath), and these will bring out the gathering (Fig. 3) of neutrophils.Yet neutrophils can not be removed and be present in residue, fragment and mikrobe throw out hidden area, that hardened.This regional neutrophils is assembled colony will cause the stomatitis spot, discharge inflammatory factor and destructive material.Oral cavity mucous membrane tissue folds and the formation of more stomatitis spots in this zone, will cause stomatitis knot (knotitis), thereby induce the visible inflammatory symptom, like stomatocace or ulcer sampling point.Along with the growth of stomatitis knot, inflammation will further increase the weight of, thereby cause oral disease, like the appearance of periodontitis.Being present in residue, fragment and mikrobe throw out hidden area, that hardened is to bring out the source that neutrophils is assembled, caused inflammatory reaction.
Therefore, collectable neutrophils number is represented in intraoral level of inflammation can be by the oral cavity.Obtaining intraoral neutrophils index can help the oral cavity level of inflammation is judged.The number of these neutrophils cells can calculate at microscopically after carrying out specific stain; Also can pass through the concentration representative of the specific component (or product) of these neutrophils cells of calculating.
According to embodiment of the present invention, the phraseology of personalized oral inflammation index (Ii) is provided, be the growth multiple of this individuality by the neutrophils cell count due to the inflammation or its specific component (or product) concentration.The calculation formula of this inflammation index is following:
I
i＝n
t/n
h
I wherein
iOral inflammation index for personalization; n
tBe neutrophils cell number or its specific component (or product) concentration of from the inflammation detected object, obtaining; n
hNeutrophils cell number or its specific component (or product) concentration under the health condition of NIP, obtained for this detected object.Detected object can be the mankind or other mammal.
In addition, the present invention also provides comparative oral inflammation index (I
c) phraseology, be neutrophils cell count or its specific component (or product) concentration of inflammation detected object, with respect to the health of NIP growth multiple with reference to the person.The calculation formula of this inflammation index is following:
I
c＝n
t/n
r
I wherein
cBe comparative oral inflammation index; n
tBe neutrophils cell number or its specific component (or product) concentration of from the inflammation detected object, obtaining; n
rBe the health of NIP neutrophils cell number or its specific component (or product) concentration with reference to the person.Detected object can be the mankind or other mammal.Among the embodiment therein, detected object is human; The health of NIP is with reference to person's neutrophils cell number (n
r) be 10,000.
Can be clear and definite be, the oral inflammation index is the growth multiple of neutrophils cell count or its specific component (or product) concentration, this can reflect degree of inflammation to a certain extent, helps doctor and patient further to understand this oral inflammation problem.
When detecting the oral inflammation index, the efficient that neutrophils cell number or its specific component (or product) discharge from the oral cavity receives used sampling damping fluid and condition effect.The invention discloses sampling damping fluid, program and the condition that can produce consistence and repeatable result, will partly carry out detailed elaboration in embodiment.
Through further investigation; The inventor has disclosed the oral inflammation level and oral cavity neutrophils cell quantity is closely related; And on the basis of this discovery; The invention provides a kind of through detecting oral cavity inflammation index in the oral cavity auxiliary judgment inflammation degree methods, and provide this method involved related prods.
At first; The invention provides a kind of novel solutions that is used in the Mammals oral cavity, discharging the neutrophils cell; Adopt the sampling solution of this solution, can discharge neutrophils from the oral cavity, and not injure human body as neutrophils cell in the inclusion of oral cavity; And can fix the polymorphic nucleus characteristic that keeps neutrophils, can guarantee to obtain to have consistence and repeatable result;
Secondly,, the invention provides the oral inflammation exponential method that detects based on above-mentioned solution and the test kit that comprises this solution, through measuring the quantity of neutrophils cell in the oral cavity, but indirect detection with estimate Mammals oral inflammation level.
Description of drawings
Below, specify embodiments of the invention in conjunction with accompanying drawing, wherein:
Fig. 1 is ulcer sampling point and cleanout tool thereof, wherein 1A: ulcer appears near the palatine tonsil; 1B: a custom-designed instrument " removes scorching brush ", and it can be used for removing the inflammation source that is embedded in the oral cavity; 1C: a ulcer sampling point appears near the mucous membrane gum intersection; 1D: the ulcer sampling point cleans the back recovery from illness.
Fig. 2 is stomatitis spot and neutrophils, wherein 2A: after removing scorching brush cleaning, discharge the brown secretory product (shown in the arrow) of coming from a ulcer sampling point; 2B: brown secretory product is with T ü rk ' s solution-dyed, and this secretory product contains a large amount of neutrophils, (arrow) that swill is seemingly relevant; 2C: the cell in the secretory product has typical polymorphic nucleus characteristic; 2D: sample is observed under fluorescent microscope with SP 15 Lemon Yellow (Acridineorange) dyeing.
The model of Fig. 3 for being inflamed in the oral cavity and proposing.
Embodiment
Below in conjunction with concrete embodiment, and comparable data describes in further detail the present invention.Should be understood that these embodiment just in order to demonstrate the invention, but not limit scope of invention by any way.
In following embodiment, various processes and the method do not described in detail are ordinary methods as known in the art.The source of agents useful for same, trade(brand)name and be necessary to list its moity person indicate when occurring first that all used thereafter identical reagent is like no specified otherwise, and is all identical with the content of indicating first.
Embodiment 1: to the detection of the neutrophils cell quantity that discharges in the oral cavity
Tested among this embodiment and accepted the intraoral neutrophils cell number of general patient (calling " No. two patients " in the following text) before and after 10 months that the ulcer sampling point is removed patient's (calling " patient " in the following text) of treatment and do not accepted corresponding treatment.
A said patient 45 years old suffers from 15 years periodontitis, is attended by once in a while halitosis and stomatocace; No. two patients 30 years old suffer from stomatocace and halitosis often take place.
In on January 1st, 2010 and on October 2nd, 2010 patient and No. two patients have been carried out the detection of oral cavity neutrophils cell number respectively, it is following specifically to detect step:
1, the neutrophils cell in the collection oral cavity is gargled with an amount of sampling damping fluid, with the neutrophils cell in the delivering oral.Adopt 15 milliliters sampling damping fluid to carry out the oral cavity of washing one's face and rinsing one's mouth for twice, each 30 seconds, from mouth, discharge the neutrophils cell, with the collutory sample mixing of gained 30 milliliters sample.This sampling damping fluid is 0.4 gram/L Repone K, 0.06 gram/L anhydrous potassium dihydrogenphosphate, 8.0 gram/L sodium-chlor, the anhydrous binary sodium phosphate of 0.04788 gram/L, 1.0 gram/LD-glucose and the phenol red sodium of 0.011 gram/L;
2, with stationary liquid with the neutrophils cell fixation.Adopt 2.5% formaldehyde fixed neutrophils cell, under 4 degrees centigrade of environment, store then, until use.
If 3 neutrophils cell concns are on the low side, need it be concentrated, so that accurately calculate.In this concrete embodiment, under the centrifugal rotational speed of 20g, concentrate the neutrophils cell.
4, after carrying out specific stain, detect and calculate the neutrophils cell count.In this concrete embodiment, the neutrophils cell (AO) is dyeed by 1.6% SP 15 Lemon Yellow (Aridine orange), under fluorescent microscope, observes and counts then.
The neutrophils cell count also can be reflected by the peculiar composition of this cell (or product) concentration; When taking a sample, can collect this distinctive composition (or product), carry out quantitative analysis then, measure its concentration in the oral cavity.The neutrophils cell endemic element of being measured (or product) possibly be protein, DNA, ribonic acid or other is at the peculiar composition of its cell (or product).The detection and the quantitative analysis of these endemic elements (or product) can be undertaken by the existing method of knowing.In scientific and technical literature, existing a lot of methods are used for detecting protein, DNA, ribonic acid and other compound.Such as ELISA can detect protein; PCR can detect DNA; Specific probe hybridization can detect RNA; Micromolecular compound can be detected or the like by HPLC or chromatography of gases.
Detected result sees the following form 1.
Table 1, a patient and No. two patients' neutrophils cell count and oral inflammation exponential are relatively
As described in Table 1, a patient has accepted to remove the treatment of ulcer sampling point, and No. two patients do not accept corresponding treatment.A patient suffers from halitosis and stomatocace once in a while when taking a sample on January 1st, 2010, the neutrophils cell number that discharged this moment is 25 * 10
4Surprisingly, when same patient took a sample after 9 months (on October 2nd, 1), this patient also no longer suffers from any halitosis and stomatocace, and the neutrophils cell number that discharged this moment is 1 * 10
4On the other hand, the neutrophils cell count that from No. two patient's mouths, is discharged, almost consistent in twice different sampling, even be separated by sample time 9 months.No. two patients continue often suffer from halitosis and stomatocace.These researchs show that the neutrophils cell count that from the oral cavity, discharges can reflect degree of inflammation in the oral cavity.
Embodiment 2: calculate the oral cavity according to oral cavity neutrophils cell count detected result among the embodiment 1
Inflammation index
Present embodiment has carried out the oral inflammation exponential according to oral cavity neutrophils cell count detected result among the embodiment 1 and has calculated.Described in table 1, patient has accepted the ulcer sampling point and has removed treatment, on January 1st, 2010 (promptly before the treatment), and collected neutrophils cell (n in this patient's mouth
t) be: 25X10
4At 2010 2 days on the 10th (i.e. treatment after), collected neutrophils cell count (n from this patient's oral cavity
h) be: 1X10
4, that is to say that this patient's neutrophils cell count has reduced 25 times.Can instead release the personalized oral inflammation index (I of this patient before treatment thus
i) be: 25X10
4/ 1X10
4=25.And this patient the treatment after, its personalized oral inflammation index (I
i) be: 1X10
4/ 1X10
4=1.
An above-mentioned patients undergo ulcer sampling point is removed after the treatment, and the health that can be designated as NIP is with reference to the person.After the treatment, this person's neutrophils cell count can be appointed as the neutrophils cell number (n of the health of NIP with reference to the person
r), promptly 10,000.Can extrapolate the comparative oral inflammation index in the table 1 thus.
No. two patient does not accept any treatment, and obvious variation does not take place this patient's neutrophils cell number.Neutrophils cell number (the n that this patient is obtained under the health condition of NIP
h) be unknown.This patient's personalized oral inflammation index (I
i) also be unknown.
Embodiment 3: the neutrophils cell counting and the comparative oral inflammation of different detected objects refer to
The detected result of number relatively
Adopt the method identical with 2 from 9 different detection objects, to collect the neutrophils cell, to its comparative oral inflammation index (I with embodiment 1
c) detect, its result lists in the following table 2.
The oral healthy condition of table 2, nine detected objects and oral inflammation index
Neutrophils cell number (n as described in Table 2, as from detected object, to obtain
t) and comparative oral inflammation index (I
c) numerical value and age do not have direct relation; It is encouraging that these numerical value can demonstrate the health problem in the oral cavity." the n of No. 5 detected objects
t" and " I
c" value the highest, this detected object often receives the puzzlement of halitosis and stomatocace." the n of No. 3 detected objects
t" and " I
c" value second height, this detected object is a smoker.Then, " the n of No. 3 detected objects
t" and " I
c" value also higher, this detected object once suffered from the carious tooth disease." the n of other detected object
t" and " I
c" value is all relative lower, these detected objects do not have tangible oral health issue.These researchs are proof once more, these " n
t" and " I
c" numerical value can the secondary proof oral inflammation level.
Because genetic different among the crowd, the neutrophils cell count between the different crowd is difference to some extent.In the variation of same intraindividual neutrophils cell, more can truly reflect intraoral inflammation degree.So, personalized oral inflammation index (I
i) understanding of inflammation is had reference value, but this exponential obtains and needs this individuality to carry out corresponding ulcer sampling point to remove treatment.Therefore under many circumstances, the tester can only compare the health of detected object and NIP with reference to the person, obtain its comparative oral inflammation index (I
c).For convenience of calculation, the health of NIP is with reference to person's neutrophils cell number (n
t) answer value be mention in above-mentioned 10000.
Should be noted that the oral inflammation index is the growth multiple of neutrophils cell count or its specific component (or product) concentration, this can reflect degree of inflammation, makes doctor and detected object can directly understand the seriousness with interpretation problems.
The disclosed instance parameter of the present invention can change to some extent, but does not influence inventive concept involved in the present invention, does not therefore limit to claim involved in the present invention.
Claims (19)
1. the solution of the interior neutrophils cell of delivering oral is characterized in that, said solution comprises salt ion and the acid ion that is used for neutrophils cell in the delivering oral;
Preferably, said salt ion and acid ion are provided by compound, and said compound is selected from Repone K, one or more of sodium-chlor and sodium phosphate.
2. solution as claimed in claim 1 is characterized in that, said solution can further comprise the compound that is used to provide cell institute energy requirement;
Preferably, provide the compound of said cell institute energy requirement to be: D-glucose.
3. according to claim 1 or claim 2 solution is characterized in that said solution can further comprise and be used to regulate the pH value of this solution and the compound of shock-absorbing capacity;
Preferably, said compound is selected from anhydrous potassium dihydrogenphosphate, one or more of sodium phosphate and anhydrous phosphoric acid hydrogen two ammoniums.
4. like each described solution among the claim 1-3, it is characterized in that said solution can further comprise and be used for painted compound;
Preferably, said compound is: phenol red sodium.
5. like each described solution among the claim 1-4; It is characterized in that; Said solution comprises: 0.01-10.00 gram/L Repone K, 0.01-10.00 gram/L anhydrous potassium dihydrogenphosphate, 0.01-10.00 gram/L sodium-chlor; 0.01-10.00 the anhydrous binary sodium phosphate of gram/L, 0.01-10.00 gram/L D-glucose and the phenol red sodium of 0.01-0.02 gram/L;
Preferably, said solution comprises 0.4 gram/L Repone K, 0.06 gram/L anhydrous potassium dihydrogenphosphate, 8.0 gram/L sodium-chlor, the anhydrous binary sodium phosphate of 0.04788 gram/L, 1.0 gram/L D-glucose and the phenol red sodium of 0.011 gram/L.
6. like each described solution among the claim 1-5, it is characterized in that said solution is the aqueous solution, spirituous solution or glucose solution;
Preferably, the ethanol concn in the said spirituous solution is 1-70%;
Preferably, the glucose concn in the said glucose solution is 0.1-15%.
7. a test kit that is used to detect neutrophils cell quantity in the oral cavity is characterized in that, said test kit comprises like each described solution among the claim 1-6.
8. test kit as claimed in claim 7 is characterized in that, said test kit further comprises:
The fixing agent that is used for the fixing said neutrophils cell of specificity; And/or
Be used for said neutrophils cell is carried out the staining agent of specific stain;
Preferably, said fixing agent is a formaldehyde;
Preferably, said staining agent is a SP 15 Lemon Yellow.
9. like claim 7 or 8 described test kits, it is characterized in that said test kit further comprises special composition or the required detection reagent of product that detects the neutrophils cell; Preferentially, said specific component is: neutrophil elastase, cathepsin G, myeloperoxidase, one or more of Lf lactoferrin and gelatinase; Its detection reagent comprises required antibody and the relevant detection reagent thereof of this differential protein of detection; Its detection reagent is the required reagent of integrated enzyme reaction.
10. be used for detecting the application of the product of neutrophils cell quantity in the oral cavity like each described solution among the claim 1-6 or like each described test kit among the claim 7-9 in preparation.
11. one kind is detected Mammals oral inflammation exponential method, it is characterized in that said method comprises: the quantity that detects neutrophils cell in the sample that comprises said Mammals oral cavity inclusion;
Preferably, said oral inflammation index is personalized oral inflammation index or comparative oral inflammation index;
Preferably, said Mammals is the people.
12. method as claimed in claim 11 is characterized in that, said method comprises:
1) said Mammals is gargled with an amount of solution, thereby obtains to comprise the solution of said Mammals oral cavity inclusion;
2) use the fixing neutrophils cell in the solution that step 1) obtains of fixing agent;
3) using the staining agent will be through step 2) fixed neutrophils cell dyes and counts;
Preferably, in step 2) preceding, the centrifugal said solution that comprises Mammals oral cavity inclusion of elder generation; Further preferably, said centrifugal speed is 20-100g, and the time is 10-30 minute; More preferably, said centrifugal speed is 20g, and the time is 10 minutes.
13. method as claimed in claim 12 is characterized in that, the solution in the said step 1) is like each described solution among the claim 1-6;
Preferably, the fixing agent said step 2) is a formaldehyde; Further preferably, said fixing agent is 2.5% formaldehyde;
Preferably, the staining agent in the said step 3) is a SP 15 Lemon Yellow; Further preferably, said staining agent is 1.6% SP 15 Lemon Yellow.
14. like each described method among the claim 10-13; It is characterized in that; Said method further comprises through the quantity of detected neutrophils cell in the said sample and the quantity of Mammals to be detected intraoral neutrophils cell under state of health are compared, calculates the oral inflammation index of said mammiferous personalization.
15. like each described method among the claim 10-14; It is characterized in that; Said method further comprises through the quantity of detected neutrophils cell in the said sample and the Mammals under the state of health quantity with reference to the intraoral neutrophils cell of person is compared, calculates said mammiferous comparative oral inflammation index;
Preferably, the Mammals under the said state of health is 10000 with reference to the quantity of the intraoral neutrophils cell of person.
16. one kind is detected Mammals oral inflammation exponential method; It is characterized in that; Said method comprises: detect the content of neutrophils cell-specific composition in the sample comprise said Mammals oral cavity inclusion or product, said specific component is selected from neutrophil elastase, cathepsin G; Myeloperoxidase, one or more of Lf lactoferrin and gelatinase;
Preferably, said oral inflammation index is personalized oral inflammation index or comparative oral inflammation index;
Preferably, said Mammals is the people.
17. method as claimed in claim 16; It is characterized in that; Said method further comprises through the content of detected neutrophils cell-specific composition or product in the said sample and the Mammals to be detected content of intraoral neutrophils cell-specific composition or product under state of health is compared, calculates the oral inflammation index of said mammiferous personalization.
18. method as claimed in claim 16; It is characterized in that; Said method further comprises through the content of detected neutrophils cell-specific composition or product in the said sample and the Mammals under the state of health content with reference to person's intraoral neutrophils cell-specific composition or product is compared, calculates said mammiferous comparative oral inflammation index.
19. one kind is detected the method that the Mammals oral hygiene is improved situation, it is characterized in that said method comprises:
Before the Mammals oral hygiene is improved,, detect Mammals oral inflammation index respectively, relatively improve before then and oral inflammation index afterwards according to each said method among the claim 10-18 with afterwards;
Preferably, said oral inflammation index is personalized oral inflammation index or comparative oral inflammation index;
Preferably, said Mammals is the people.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201010588508XA CN102533933A (en) | 2010-12-15 | 2010-12-15 | Method, reagent and kit for detecting oral inflammatory index, and applications thereof |
| PCT/CN2011/000226 WO2011097962A1 (en) | 2010-02-12 | 2011-02-12 | Method for detecting and tool for clearing stomatitis spots and stomatitis nodes |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201010588508XA CN102533933A (en) | 2010-12-15 | 2010-12-15 | Method, reagent and kit for detecting oral inflammatory index, and applications thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN102533933A true CN102533933A (en) | 2012-07-04 |
Family
ID=46341962
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201010588508XA Pending CN102533933A (en) | 2010-02-12 | 2010-12-15 | Method, reagent and kit for detecting oral inflammatory index, and applications thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102533933A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105287646A (en) * | 2014-06-07 | 2016-02-03 | 潘申权 | Solution preparation for detecting and relieving or eliminating oral inflammation, combination product and application thereof |
| CN106461658A (en) * | 2014-06-05 | 2017-02-22 | 高露洁-棕榄公司 | Assay for oral inflammation |
| CN112638314A (en) * | 2018-07-02 | 2021-04-09 | 佩里奥诺瓦姆有限责任公司 | Device and method for oral hygiene |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20080085533A1 (en) * | 2006-10-05 | 2008-04-10 | Michael Glogauer | Method and apparatus for non -invasive rinse diagnosis and monitoring of periodontal diseases |
| CN101185001A (en) * | 2005-03-31 | 2008-05-21 | C2诊断公司 | Method for analyzing a blood sample and apparatus and reagent for implementing said method |
| CN101443660A (en) * | 2006-03-15 | 2009-05-27 | 综合医院公司 | Devices and methods for detecting cells and other analytes |
| CN101785710A (en) * | 2010-02-12 | 2010-07-28 | 潘申权 | Tool for clearing stomatitis spots and stomatitis nodes, and application thereof |
| CN102559836A (en) * | 2010-12-15 | 2012-07-11 | 潘申权 | Reagent for detecting impact index of intimacy degree, kit, method and application thereof |
-
2010
- 2010-12-15 CN CN201010588508XA patent/CN102533933A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101185001A (en) * | 2005-03-31 | 2008-05-21 | C2诊断公司 | Method for analyzing a blood sample and apparatus and reagent for implementing said method |
| CN101443660A (en) * | 2006-03-15 | 2009-05-27 | 综合医院公司 | Devices and methods for detecting cells and other analytes |
| US20080085533A1 (en) * | 2006-10-05 | 2008-04-10 | Michael Glogauer | Method and apparatus for non -invasive rinse diagnosis and monitoring of periodontal diseases |
| CN101785710A (en) * | 2010-02-12 | 2010-07-28 | 潘申权 | Tool for clearing stomatitis spots and stomatitis nodes, and application thereof |
| CN102559836A (en) * | 2010-12-15 | 2012-07-11 | 潘申权 | Reagent for detecting impact index of intimacy degree, kit, method and application thereof |
Non-Patent Citations (7)
| Title |
|---|
| ASHKENAZI M,DENNISON DK ET AL: "A new method for isolation of salivary neurophils and determination of their functional activity", 《J DEN RES》 * |
| BENDER JS, THANG ET AL: "Novel rinse assay for the quantification of oral neutrophils and the monitoring of chronic periodontal disease", 《J PERIODONT RES》 * |
| R.I.弗雷谢尼著 章静波等译: "《动物细胞培养——基本技术指南(第四版)》", 31 December 2004, 科学出版社 * |
| SIGMA-ALDRICH: "Hanks" balanced salts [HBSS] without sodium bicarbonate", 《PRODUCT NUMBER H6136》 * |
| 司徒镇强、吴军正: "《细胞培养》", 31 January 2007, article "附录IV 细胞培养常用的溶液", pages: 330-331 * |
| 司徒镇强、吴军正: "《细胞培养》", 31 January 2007, 世界图书出版西安公司 * |
| 陈汉程: "白血球在炎症组织内的活动", 《中国兽医杂志》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106461658A (en) * | 2014-06-05 | 2017-02-22 | 高露洁-棕榄公司 | Assay for oral inflammation |
| CN105287646A (en) * | 2014-06-07 | 2016-02-03 | 潘申权 | Solution preparation for detecting and relieving or eliminating oral inflammation, combination product and application thereof |
| CN112638314A (en) * | 2018-07-02 | 2021-04-09 | 佩里奥诺瓦姆有限责任公司 | Device and method for oral hygiene |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Lima et al. | Saliva: reflection of the body | |
| Motamayel et al. | Saliva as a mirror of the body health | |
| Nriagu et al. | Lead levels in blood and saliva in a low-income population of Detroit, Michigan | |
| CN104620110A (en) | Analysis of saliva proteome for biomarkers of gingivitis and periodontitis using ft-icr-ms/ms | |
| Koppolu et al. | Estimate of CRP and TNF-alpha level before and after periodontal therapy in cardiovascular disease patients | |
| CN102533933A (en) | Method, reagent and kit for detecting oral inflammatory index, and applications thereof | |
| Giannobile et al. | Glycosaminoglycans and periodontal disease: analysis of GCF by safranin O | |
| RU2525437C1 (en) | Method for determining oxidative protein modification in substance pool of average molecular weight in blood serum, plasma, erythrocyte, and in urine | |
| CN102559836A (en) | Reagent for detecting impact index of intimacy degree, kit, method and application thereof | |
| JP6800470B2 (en) | Periodontitis test kit and test method using miRNA as a marker | |
| CN107271655A (en) | A kind of kit and method for detecting urine excretion body load miRNAs | |
| RU2702736C2 (en) | Examination in oral cavity inflammation | |
| CN108929903A (en) | Marker and kit for chronic periodontitis screening, diagnosis, therapeutic evaluation | |
| Sitaru et al. | Correlation between the salivary level of alpha-amylase and the risk for dental caries in young permanent teeth | |
| BR9911156A (en) | Method and system for the production and collection of faeces induced by washing (fil) for chemical and biological tests of cells | |
| US20140273280A1 (en) | Method for diagnosing chronic sinusitis | |
| Ochmański et al. | Analysis of chemical composition of urinary stones | |
| Saulin et al. | Influence of the local and systemic oxidative stress on periodontitis: role of antioxidant therapy | |
| Ramya et al. | Salivary Biomarkers for Periodontal Diseases and Personalized Monitoring-A Critical Review | |
| Lamberts et al. | Fibronectin levels of unstimulated saliva from naval recruits with and without chronic inflammatory periodontal disease | |
| RU2473915C1 (en) | Method of determining degree of severity of periodontitis | |
| RU2793526C1 (en) | Method for diagnosing intolerance to acrylic structural materials of removable lamellar dentures in elderly and senile patients | |
| Fokkema et al. | A possible association of α1‐antitrypsin deficiency with the periodontal condition in adults | |
| CN105287646A (en) | Solution preparation for detecting and relieving or eliminating oral inflammation, combination product and application thereof | |
| Cahaya et al. | Correlation between calcium and phosphate levels to calculus accumulation on coronary heart disease patients |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20120704 |
|
| RJ01 | Rejection of invention patent application after publication |