CN206470277U - Nasopharyngeal carcinoma precisely diagnoses fully-automated synthesis equipment - Google Patents
Nasopharyngeal carcinoma precisely diagnoses fully-automated synthesis equipment Download PDFInfo
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Abstract
A kind of nasopharyngeal carcinoma precisely diagnoses fully-automated synthesis equipment, including the full-automatic fluorescence immunity analyzers of HELIOS and instant titre quantitative interpretation device；Epstein-Barr virus VCA/EA IgA cellular immunofluorescence detection reagents main component includes cell sheet, the anti-FITC IgA of fluorescence two, positive control quality-control product, negative control quality-control product, mountant, PBS pulvis and cover glass；The full-automatic fluorescence immunity analyzers of HELIOS are provided with camera system, and camera system is connected with instant titre quantitative interpretation device；The full-automatic fluorescence immunity analyzers of HELIOS carry out automatic detection to sample, and the sample of detection is imaged by camera system, and the image results of sample imaging are inputted to instant titre quantitative interpretation device, and titre results are obtained by instant titre quantitative interpretation device.The utility model automatic detection simultaneously carries out result interpretation, with testing result precision it is high, it is easy to detect the characteristics of.
Description
Technical field
The utility model is related to accurate technical field of medical equipment, and precisely diagnosis is full-automatic for more particularly to a kind of nasopharyngeal carcinoma
Detection device.
Background technology
Nasopharyngeal carcinoma is the common malignant tumour in south China area, and nasopharyngeal carcinoma is a multifactor, multistage mistake
Journey.Epstein-Barr virus can cause nasopharyngeal epithelial cell to immortalize, so as to further deteriorate it.
In for the examination of nasopharyngeal carcinoma and clinical diagnosis research, the Epstein-Barr virus antigen more determined mainly has capsid antigen
And early antigens (EA) (VCA).Confirmation EBV VCA-IgA antibody has been studied to may occur in which before nasopharyngeal carcinoma is made a definite diagnosis 10-61 months,
Contribute to find early stage nasopharyngeal carcinoma by lasting detection EBV VCA-IgA.EA-IgA appearance generally points out body Epstein-Barr virus to answer
System starts, and has higher specificity in Nasopharyngeal Carcinoma Patients, is disadvantageous in that sensitiveness is relatively low, Clinical screening and serodiagnosis
Used cooperatively more with VCA-IgA, the accuracy rate of examination can be improved.
VCA-IgA and EA-IgA common detection method has：Enzyme linked immunosorbent assay, immunoenzyme, indirectly be immunized it is glimmering
Light method.
Enzyme linked immunosorbent assay：VCA or EA recombinant antigens are coated in microwell plate, in the antibody and micropore in serum
Antigen-reactive, forms antigen-antibody complex；ELIAS secondary antibody is added after board-washing to be reacted, and forms Ag-Ab-enzyme mark two
Anti- compound；Chromogenic substrate is added after board-washing and carries out chromogenic reaction, acid terminate liquid terminating reaction is then added.Use ELIASA
Read the OD values of reaction solution.According to the OD values of the OD values of comparative sample and positive quality control product come interpretation yin and yang attribute.Enzyme linked immunological is inhaled
Attached method sensitiveness is high, but specificity is low, easily produces false positive and false negative.Other this method belongs to sxemiquantitative, it is impossible to realize
It is precisely quantitative.
Immunoenzyme：Specific cell is fixed on slide, and the cell incubation on antibody and slide in serum is anti-
Should, form cell-antibody compound；ELIAS secondary antibody is added after developing a film, cell-antibody-ELIAS secondary antibody compound is formed；After developing a film
Add chromogenic substrate reaction；Reaction is developed a film after terminating, and mounting, and cell color situation is observed using ordinary optical microscope.Root
The yin and yang attribute of serum sample is judged according to the cell color depth.There is the defect that interference factor is more, easily judges by accident in immunoenzyme, and
Then to there is subjectivity in artificial interpretation strong again, it is difficult to carries out quality control and uniformly compares.
IIF：Specific cell is fixed on slide, the cell on antibody and slide in serum
Incubation reaction, forms cell-antibody compound；Fluorescein mark secondary antibody is added after developing a film, cell-antibody-fluorescein mark is formed
Remember secondary antibody compound；Reaction is developed a film after terminating, and mounting, utilizes fluorescence microscope cell fluorescence situation.It is glimmering according to cell
Light whether there is or the depth judges the yin and yang attribute of serum sample.IIF sensitiveness is not high, automaticity is low, subjectivity
By force, it is impossible to realize Standardized quantitative.
On the one hand the accuracy of any of the above detection method result, the limitation of automation mechanized operation degree are the quick of detection reagent
Perception, specificity are not obvious enough, on the other hand, also carry out detection operation without simple automation equipment, utilize artificial operation
The efficiency and accuracy of result are limited to, particularly artificial progress interpretation of result is easily affected by various factors.At present still
There is no complete detection architecture to carry out automated diagnostic fully-automated synthesis to Epstein-Barr virus VCA/EA-IgA cells.
Therefore, in view of the shortcomings of the prior art, providing a kind of nasopharyngeal carcinoma, precisely to diagnose fully-automated synthesis equipment existing to overcome
Technical deficiency is very necessary.
Utility model content
The purpose of this utility model is to avoid the deficiencies in the prior art part and provide a kind of nasopharyngeal carcinoma and precisely diagnose entirely
Automatic checkout equipment, the nasopharyngeal carcinoma precisely diagnoses fully-automated synthesis equipment and used to be had well to Epstein-Barr virus VCA/EA-IgA antibody
Sensitivity and Specificity detection reagent and combine the full-automatic fluorescence immunity analyzers of HELIOS and instant titre and quantitatively sentence
Read apparatus, can be achieved the automation of fluoroscopic examination, and the fluorescence results picture detected for EBV VCA/EA IgA CIF is carried out
Judge, with testing result precision it is high, it is easy to detect the characteristics of.
Above-mentioned purpose of the present utility model is realized by following technological means.
A kind of nasopharyngeal carcinoma is provided and precisely diagnoses fully-automated synthesis equipment, including uses Epstein-Barr virus VCA/EA-IgA cellular immunities
The full-automatic fluorescence immunity analyzers of HELIOS and instant titre quantitative interpretation device of luciferase assay reagent；
Epstein-Barr virus VCA/EA-IgA cellular immunofluorescence detection reagents main component includes cell sheet, fluorescence secondary antibody FITC-
IgA, positive control quality-control product, negative control quality-control product, mountant, PBS pulvis and cover glass；
The full-automatic fluorescence immunity analyzers of HELIOS are provided with camera system, and camera system is filled with instant titre quantitative interpretation
Put connection；
The full-automatic fluorescence immunity analyzers of HELIOS carry out automatic detection to sample, and by camera system to detection
Sample is imaged, and the image results of sample imaging are inputted to instant titre quantitative interpretation device, are filled by instant titre quantitative interpretation
Put acquisition titre results.
It is preferred that, above-mentioned nasopharyngeal carcinoma precisely diagnoses fully-automated synthesis equipment, the full-automatic fluorescence immunity analyzers of HELIOS
Camera system be specially micro-photographing apparatus, micro-photographing apparatus is connected with the instant titre quantitative interpretation device.
It is preferred that, above-mentioned nasopharyngeal carcinoma precisely diagnoses fully-automated synthesis equipment, and real-time and quantification interpretation device is included photo
Be converted into gray scale picture gradation conversion device, by gray scale picture carry out black white reverse inversion device, carry out unit area fluorescence it is strong
Spend the calculator calculated and the analyzer that titre numerical value is obtained according to unit area fluorescence intensity.
It is preferred that, above-mentioned nasopharyngeal carcinoma precisely diagnoses fully-automated synthesis equipment, and photo is converted into 8 by gradation conversion device
Gray scale picture.
It is preferred that, above-mentioned nasopharyngeal carcinoma precisely diagnoses fully-automated synthesis equipment, and the analyzer unit area fluorescence that is stored with is strong
The numerical value of degree and titre matching list, matching list are specific as follows；
When the numerical value of unit area fluorescence intensity is less than 0.05, titre value is less than 10；
When the numerical value of unit area fluorescence intensity is less than 0.08 more than or equal to 0.05, titre value is determined as 10；
When the numerical value of unit area fluorescence intensity is less than 0.12 more than or equal to 0.08, titre value is determined as 20；
When the numerical value of unit area fluorescence intensity is less than 0.16 more than or equal to 0.12, titre value is determined as 40；
When the numerical value of unit area fluorescence intensity is less than 0.19 more than or equal to 0.16, titre value is determined as 80；
When the numerical value of unit area fluorescence intensity is less than 0.22 more than or equal to 0.19, titre value is determined as 160；
When the numerical value of unit area fluorescence intensity is less than 0.24 more than or equal to 0.22, titre value is determined as 320；
When the numerical value of unit area fluorescence intensity is less than 0.3 more than or equal to 0.24, titre value is determined as 640；
When the numerical value of unit area fluorescence intensity is more than 0.3, titre value is judged to being more than 640.
Nasopharyngeal carcinoma of the present utility model precisely diagnoses fully-automated synthesis equipment, including uses Epstein-Barr virus VCA/EA-IgA cells
The full-automatic fluorescence immunity analyzers of HELIOS and instant titre quantitative interpretation device of immunofluorescence detection agent；Epstein-Barr virus VCA/
EA-IgA cellular immunofluorescence detection reagents main component include cell sheet, fluorescence secondary antibody FITC-IgA, positive control quality-control product,
Negative control quality-control product, mountant, PBS pulvis and cover glass；The full-automatic fluorescence immunity analyzers of HELIOS are provided with shooting system
System, camera system is connected with instant titre quantitative interpretation device；The full-automatic fluorescence immunity analyzers of HELIOS are carried out certainly to sample
Dynamicization is detected, and the sample of detection is imaged by camera system, and the image results of sample imaging, which are inputted to instant titre, to be quantified
Interpretation device, titre results are obtained by instant titre quantitative interpretation device.The nasopharyngeal carcinoma precisely diagnoses fully-automated synthesis equipment
It is using the detection reagent to Epstein-Barr virus VCA/EA-IgA antibody with good Sensitivity and Specificity and complete certainly with reference to HELIOS
Dynamic fluorescence immunity analyzer and instant titre quantitative interpretation device, can be achieved the automation of fluoroscopic examination, and for EBV
VCA/EA IgA CIF detection fluorescence results picture judged, with testing result precision it is high, it is easy to detect the characteristics of.
Brief description of the drawings
The utility model is elaborated with reference to accompanying drawing, but the content in accompanying drawing is not constituted to limit of the present utility model
System.
Fig. 1 is the schematic diagram that the utility model nasopharyngeal carcinoma precisely diagnoses fully-automated synthesis equipment.
Include in Fig. 1：
The full-automatic fluorescence immunity analyzers 100 of ELIOS,
Instant titre quantitative interpretation device 200.
Embodiment
The utility model is further described with the following Examples.
Embodiment 1.
A kind of nasopharyngeal carcinoma precisely diagnoses fully-automated synthesis equipment, as shown in figure 1, including HELIOS full-automatic fluorescence immunoassays point
Analyzer 100 and instant titre quantitative interpretation device 200.
The full-automatic fluorescence immunity analyzers of HELIOS use Epstein-Barr virus VCA/EA-IgA cellular immunofluorescence detection reagents.EB
It is right that viral VCA/EA-IgA cellular immunofluorescences detection reagent main component includes cell sheet, fluorescence secondary antibody FITC-IgA, the positive
According to quality-control product, negative control quality-control product, mountant, PBS pulvis and cover glass.
The full-automatic fluorescence immunity analyzers of HELIOS are provided with camera system, and camera system is filled with instant titre quantitative interpretation
Put connection.The full-automatic fluorescence immunity analyzers of HELIOS carry out automatic detection to sample, and by camera system to detection
Sample is imaged, and the image results of sample imaging are inputted to instant titre quantitative interpretation device, are filled by instant titre quantitative interpretation
Put acquisition titre results.
Specifically, the nasopharyngeal carcinoma precisely diagnoses fully-automated synthesis equipment, the full-automatic fluorescence immunity analyzers of HELIOS are set
There is micro-photographing apparatus, micro-photographing apparatus is connected with the instant titre quantitative interpretation device.
Specifically, real-time and quantification interpretation device includes photo being converted into the gradation conversion device of gray scale picture, by gray-scale map
Piece carries out the inversion device of black white reverse, carries out the calculator of unit area fluorescence intensity calculating and strong according to unit area fluorescence
Degree obtains the analyzer of titre numerical value.Real-time and quantification interpretation device obtains the titre results of picture by the following method,
A, gray scale picture is converted into by photo, and photo is preferably converted into the gray scale picture of 8；
B, the gray scale picture that step A is obtained carries out black white reverse, and the picture after being inverted is named as reversion picture；
C, carries out optical density correction to reversion picture, excludes after non-specific particulate fluorescence, unit of account area fluorescence is strong
Degree；
D, titre numerical value is obtained according to unit area fluorescence intensity.
The numerical value of unit area fluorescence intensity and titre matching list specifically, analyzer is stored with, matching list are specific as follows；
When the numerical value of unit area fluorescence intensity is less than 0.05, titre value is less than 10；
When the numerical value of unit area fluorescence intensity is less than 0.08 more than or equal to 0.05, titre value is determined as 10；
When the numerical value of unit area fluorescence intensity is less than 0.12 more than or equal to 0.08, titre value is determined as 20；
When the numerical value of unit area fluorescence intensity is less than 0.16 more than or equal to 0.12, titre value is determined as 40；
When the numerical value of unit area fluorescence intensity is less than 0.19 more than or equal to 0.16, titre value is determined as 80；
When the numerical value of unit area fluorescence intensity is less than 0.22 more than or equal to 0.19, titre value is determined as 160；
When the numerical value of unit area fluorescence intensity is less than 0.24 more than or equal to 0.22, titre value is determined as 320；
When the numerical value of unit area fluorescence intensity is less than 0.3 more than or equal to 0.24, titre value is determined as 640；
When the numerical value of unit area fluorescence intensity is more than 0.3, titre value is judged to being more than 640.
The nasopharyngeal carcinoma precisely diagnoses fully-automated synthesis equipment, Epstein-Barr virus VCA/EA-IgA cellular immunofluorescences detection reagent tool
There is sensitiveness high, the characteristics of specific good.The preparation process of its cell sheet is as follows,
(1) cell line is set up
(1.1) setting of nutrient solution：The culture medium based on RPMI1640, the hyclone of addition 10%~15% is adjusted
Whole nutrient solution PH initial values are 7.2~7.4.
(1.2) B95-8 cell lines and Raji cell lines are set up
B95-8 can naturally express EBV VCA, and the technique of setting up of B95-8 cell lines is：B95-8 cells are put in above-mentioned training
In nutrient solution, it is placed in 35~37 DEG C and contains 3~6%CO2Incubator in cultivate, set up the B95-8 cells of expression efficiency stable uniform
System.
Raji cell culture conditions are as B95-8 cells, but Raji cells must can express EBV through activator activation
EA.The activation technology of Raji cell lines is：Centrifuged under conditions of 500~1000rpm 5~10 minutes and collect Raji cells, used
Raji cells are resuspended in new nutrient solution, and adjust Raji cells deeps for (1~3) × 106Individual/ml, increase Sodium butyrate to end
Concentration is 0.1~1mM, and TPA is to final concentration of 10~100ng/ml for increase, is placed in 35~37 DEG C and contains 3~6%CO2Culture
12~48h is cultivated in case, that is, establishes the Raji cell lines of expression efficiency stable uniform.
(2) the homogeneity control of cell sheet
The cell control passage of smear is within 50 generations, and it is standard less than 5% to crack number with cell, with identical smear
Technique carries out smear.
The fluorescence secondary antibody FITC-IgA of the utility model dilution, positive control quality-control product, negative control quality-control product is 2~8
It can stablize 12 months under conditions of DEG C.
The effect of stability is reached by stabilizer function.Specifically, fluorescence secondary antibody FITC-IgA, positive control quality-control product
Or the dilution of negative control quality-control product contains stabilizer, the control of the pH value of the dilution containing stabilizer 7.2 to 7.3 it
Between.
Using dilution parts by weight as 1, stabilizer contains the composition of following percentage by weight：
Glycine 1%~5%,
Extra large bath sugar 1%~5%,
BSA 1%~5%,
PBS 1%~3%.
Specifically, by weight percentage, mountant contains：
Glycerine 10%~30%；
Fluorescence protective agent 3~8%；
PBS 50%~90%.
Epstein-Barr virus VCA/EA-IgA cellular immunofluorescence detection reagents, finished product structure is to have 12 holes on a slide
The structure of hydrophobic circular hole, the hydrophobic circular hole in 12 holes is set in two row, and each column is 6.It is preferred that, the hydrophobic circular hole in 12 holes is arranged in two rows six
Matrix is arranged.There is the proprietary two-dimension identification code of project in order to recognize on accurate and automation application, the hydrophobic slide in 12 holes, fit
For being matched with the full-automatic fluorescence immunity analyzers of HELIOS, and can the instant titres of supporting EBV VCA/EA-IgA CIF quantitatively sentence
Read apparatus carries out real-time quantitative interpretation so that automatic monitoring is possibly realized.
Epstein-Barr virus VCA/EA-IgA cellular immunofluorescence detection reagents of the present utility model, newly improving two kinds can through activation
Stability and high efficiency expresses the cell line B95-8 and Raji of antigen, while having been done to cell sheet, fluorescence secondary antibody and quality-control product at stability
Reason and optimization, make the Sensitivity and Specificity of reagent greatly improve.
By adding stabilizer to cell sheet, fluorescence secondary antibody and EBV VCA/EA IgA quality-control products, make the effect phase of reagent big
It is big to increase.The term of validity compared to product in the market improves at least one times.
The finished product of EBV VCA/EA IgA CIF detection reagents is used in full-automatic fluorescence immunity analyzer
(HELIOS), realize that operating process is automated.And the painting blade technolgy and uniformity controlling of optimization so that cell sheet difference between batch subtracts
It is few, apply in the instant titre quantitative interpretation devices of EBV VCA/EA IgA CIF, more accurately quantitative result can be drawn, realize complete
The comparativity of state's laboratory monitoring.
Using equipment system of the present utility model, detected, comprised the following steps that：
1) washing lotion：PBS pulvis is diluted to working concentration with purified water.
2) dilute serum：Test serum makees 1 with washing lotion：10 dilutions are stand-by.
3) it is loaded：Yin and yang attribute control and the serum diluted, 35 μ L/ holes are separately added into the reacting hole of antigen slide.
4) it is incubated：By (21~25 DEG C) incubation 30min of antigen slide room temperature.
5) wash：Take out and expand former piece, gently reacting hole is rinsed 2 times from side with washing lotion, then antigen slide is placed in washed in cylinder
Soak 5min.Antigen slide is taken out, the raffinate around reacting hole is blotted with blotting paper.
6) fluorescence secondary antibody is added：Fluorescence secondary antibody (FITC-IgA), 25 μ L/ holes are added in the reacting hole of antigen slide.
7) it is incubated：Antigen slide is incubated at room temperature 30min.
8) wash：Repeat step 5.
9) mounting：About 10 μ L mountant is added dropwise per reacting hole.
10) result is observed：The microscopic imaging device automatic camera that the full-automatic fluorescence immunity analyzers of HELIOS are carried, immediately
Titre quantitative interpretation device is automatic to carry out titre quantitative interpretation to result picture, draws titre results.
The nasopharyngeal carcinoma precisely diagnoses fully-automated synthesis equipment and used has well quick to Epstein-Barr virus VCA/EA-IgA antibody
Perceptual and specific detection reagent simultaneously combines the full-automatic fluorescence immunity analyzers of HELIOS and instant titre quantitative interpretation dress
Put, the automation of fluoroscopic examination can be achieved, and judged for the EBV VCA/EA IgA CIF fluorescence results pictures detected,
With testing result precision it is high, it is easy to detect the characteristics of.
Embodiment 2.
A kind of nasopharyngeal carcinoma precisely diagnoses fully-automated synthesis equipment, and further feature is same as Example 1, and difference is：
The stabilizer of Epstein-Barr virus VCA/EA-IgA cellular immunofluorescence detection reagents contains the composition of following percentage by weight：
Glycine 2%~3%,
Extra large bath sugar 1%~4%,
BSA 2%~3%,
PBS 2%~2.5%.
In the utility model it is new improve two kinds through activation can stability and high efficiency express the cell line B95-8 and Raji of antigen,
Disposal of stability and optimization have been done to cell sheet, fluorescence secondary antibody and quality-control product simultaneously, made the Sensitivity and Specificity of reagent big
It is big to improve.
By adding stabilizer to cell sheet, fluorescence secondary antibody and EBV VCA/EA IgA quality-control products, make the effect phase of reagent big
It is big to increase.The term of validity compared to product in the market improves at least one times.
The finished article design of EBV VCA/EA IgA CIF detection reagents can be used in full-automatic fluorescence immunity analyzer
(HELIOS), realize that operating process is automated.And the painting blade technolgy and uniformity controlling of optimization so that cell sheet difference between batch subtracts
It is few, apply in the instant titre quantitative interpretation systems of EBV VCA/EA IgA CIF, more accurately quantitative result can be drawn, realize complete
The comparativity of state's laboratory monitoring.
Embodiment 3.
A kind of nasopharyngeal carcinoma precisely diagnoses fully-automated synthesis equipment, and further feature is same as Example 1, and difference is：
Epstein-Barr virus VCA/EA-IgA cellular immunofluorescence detection reagents, by weight percentage, mountant contains：
Glycerine 15%~20%；
Fluorescence protective agent 5~7%；
PBS 65%~80%.
The Sensitivity and Specificity performance of reagent of the present utility model is good, has good stability, and is adapted to and automation equipment
Support the use.
Finally it should be noted that above example is only to illustrate the technical solution of the utility model rather than to this practicality
The limitation of novel protected scope, although being explained in detail with reference to preferred embodiment to the utility model, the common skill of this area
Art personnel should be appreciated that can modify or equivalent substitution to the technical solution of the utility model, without departing from this practicality
The spirit and scope of new technique scheme.
Claims (5)
1. a kind of nasopharyngeal carcinoma precisely diagnoses fully-automated synthesis equipment, it is characterised in that：Including thin using Epstein-Barr virus VCA/EA-IgA
The full-automatic fluorescence immunity analyzers of HELIOS and instant titre quantitative interpretation device of born of the same parents' immunofluorescence detection agent；
Epstein-Barr virus VCA/EA-IgA cellular immunofluorescence detection reagents main component include cell sheet, fluorescence secondary antibody FITC-IgA,
Positive control quality-control product, negative control quality-control product, mountant, PBS pulvis and cover glass；
The full-automatic fluorescence immunity analyzers of HELIOS are provided with camera system, and camera system connects with instant titre quantitative interpretation device
Connect；
The full-automatic fluorescence immunity analyzers of HELIOS carry out automatic detection to sample, and pass through sample of the camera system to detection
Imaging, the image results of sample imaging are inputted to instant titre quantitative interpretation device, are obtained by instant titre quantitative interpretation device
Obtain titre results.
2. nasopharyngeal carcinoma according to claim 1 precisely diagnoses fully-automated synthesis equipment, it is characterised in that：HELIOS is full-automatic
The camera system of fluorescence immunity analyzer is specially micro-photographing apparatus, micro-photographing apparatus and the instant titre quantitative interpretation
Device is connected.
3. nasopharyngeal carcinoma according to claim 2 precisely diagnoses fully-automated synthesis equipment, it is characterised in that：Real-time and quantification interpretation
Device includes photo being converted into the gradation conversion device of gray scale picture, gray scale picture is carried out to the inversion device of black white reverse, carried out
Calculator and the analyzer according to unit area fluorescence intensity acquisition titre numerical value that unit area fluorescence intensity is calculated, it is described
Gradation conversion device, inversion device, enter calculator and analyzer is sequentially connected.
4. nasopharyngeal carcinoma according to claim 3 precisely diagnoses fully-automated synthesis equipment, it is characterised in that：Gradation conversion device will
Photo is converted into the gray scale picture of 8.
5. nasopharyngeal carcinoma according to claim 4 precisely diagnoses fully-automated synthesis equipment, it is characterised in that：Analyzer is stored with
The numerical value of unit area fluorescence intensity and titre matching list, matching list are specific as follows；
When the numerical value of unit area fluorescence intensity is less than 0.05, titre value is less than 10；
When the numerical value of unit area fluorescence intensity is less than 0.08 more than or equal to 0.05, titre value is determined as 10；
When the numerical value of unit area fluorescence intensity is less than 0.12 more than or equal to 0.08, titre value is determined as 20；
When the numerical value of unit area fluorescence intensity is less than 0.16 more than or equal to 0.12, titre value is determined as 40；
When the numerical value of unit area fluorescence intensity is less than 0.19 more than or equal to 0.16, titre value is determined as 80；
When the numerical value of unit area fluorescence intensity is less than 0.22 more than or equal to 0.19, titre value is determined as 160；
When the numerical value of unit area fluorescence intensity is less than 0.24 more than or equal to 0.22, titre value is determined as 320；
When the numerical value of unit area fluorescence intensity is less than 0.3 more than or equal to 0.24, titre value is determined as 640；
When the numerical value of unit area fluorescence intensity is more than 0.3, titre value is judged to being more than 640.
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