CN101592662A - Hepatitis B surface antibody magnetic particle chemiluminescent immunoassay kit and preparation method thereof - Google Patents
Hepatitis B surface antibody magnetic particle chemiluminescent immunoassay kit and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a kind of hepatitis B surface antibody magnetic particle chemiluminescent immunoassay kit and preparation method thereof, this kit can detect the hepatitis B surface antibody content in the human serum quickly and accurately.Kit of the present invention comprises that the hepatitis B surface antigen bag is by the streptavidin of the mixed liquor of magnetic particle and biotin labeling hepatitis B surface antigen, horseradish peroxidase-labeled, hepatitis B surface antibody calibration object, chemical luminous substrate, concentrated cleaning solution and reaction tube.Kit of the present invention adopts the dual-antigen sandwich method principle, in conjunction with magnetic particle immunological technique, biotin-avidin and chemiluminescence immunoassay technology, it detects fast, the curve linear wide ranges, calibration object is demarcated with national linear reference product, can detect the content of human serum hepatitis B surface antibody exactly, have important clinical application value.
Description
Technical field
The present invention relates to a kind of hepatitis B surface antibody magnetic particle chemiluminescent immunoassay kit and preparation method thereof.Kit of the present invention combines magnetic particle isolation technics, biotin-avidin amplifying technique and chemiluminescence immunoassay technology.
Background technology
Hepatitis B is a kind of serious common liver diseases, arrives the millions of people in global implication.In the present global population, surpass 2,000,000,000 people and infected hepatitis type B virus (HBV) certain period in its life, wherein, about 3.5 hundred million still are chronic infection person, become the carrier of virus.Whole world three quarters of the population is lived in the district occurred frequently of infection.The acute clinical case of annual HBV surpasses 400 ten thousand, and about 25% among the carrier, just annual 1000000 people die from chronic active hepatitis, cirrhosis or primary carcinoma of liver.
After infecting HBV, the infection index of first appearance is HBsAg, occurs in 1 to 3 months after being exposed to HBV.Approximately after 1-2 month, first antibody appears, promptly at the IgM antibody (anti--HBc IgM) of HBV cAg (HBcAg), at this moment near, the activity of AST and ALT rises in the serum.When jaundice occurring, the existing HBsAg of Most patients has anti--HBc IgM again.Along with the removing of virus, anti--HBs can detect.Among the patient of less ratio, the situation that HBsAg and anti--HBs can't detect may appear, the index that can detect this moment usually be IgM anti--HBc, this situation is referred to as " core window phase ", such patient anti--HBe usually mostly is positive.After HBV infected rehabilitation, most people were anti--and HBc and anti--HBs will be lifelong positive.
The immunization method that is used to detect the hepatitis B blood serum designated object at present mainly contains enzyme immunoassay (EIA) (enzymeimmunoassay, EIA), radiommunoassay (radioimmunoassay, RIA), immunofluorescence assay (fluoroimmunoassay, FIA) and chemiluminescence immune assay (chemiluminescence immunoassay, CLIA) etc.
Therefore radiating immuning analysis technology has certain contaminative to environment, and exists instrument cost expensive because of it uses radioelement thing that serves as a mark, and sensitivity is not high, complicated operation, measurement result instability, shortcoming such as the reagent holding time is short.Enzyme immunoassay (EIA) sensitivity is low, and influence factor is more, easily causes false negative and false positive.Chemiluminescence immunoassay is a kind of than elder generation and then effective method, can make detection sensitivity reach 10
-18Mol level, and sensing range can reach 6 orders of magnitude, because the enzyme labeling thing is stable, can uses for a long time, thereby obtain increasing concern.
Immunity magnetic particle technology is to utilize the magnetic solid phase particle of the synthetic certain particle size size of macromolecular material to make carrier, with method bags such as physisorption, chemical coupling by on have the specificity affinity various immunologic active materials (antigen or antibody), making its sensitization is immune magnetic particle, has that velocity of separation is fast, efficient is high, favorable repeatability; Do not influence characteristics such as the biological character of separated cell or other biomaterial and function, adding directed movement under the action of a magnetic field, make some special composition be separated, concentrate or purifying.As a kind of general solid phase isolation methods that is used for the chemiluminescence immune assay detection technique, thereby magnetic particle can improve effective package amount conservation greatly, can widen the range of linearity of detection simultaneously, thereby effectively avoid the generation of crotch effect.Immune magnetic particle technology is combined the detection determinand with chemiluminescence immunoassay technology, can improve the sensitivity and the accuracy of detection greatly, it is a carrier with the micron order magnetic particle, carboxyl reactive group and the protein amino covalent bond of utilizing surface organic matter to provide, adopt antibody to carry out " bridging " and become immune magnetic particle, can carry out antigen, antibody response.The novel part of this technology has: (1) utilizes the paramagnetic iron particulate to be solid phase carrier, the expoeridium gene engineering antigen, increase the contact area and the substrate light-emitting area of antigen, antibody, improve the sensitivity of reaction, and adopt rotating magnetic field to make magnetic particle play beating action and separating and combining Ag-Ab and free antibodies; (2) in liquid phase reactor, use luminescence enhancer, the luminescence sites of hydrone from luminous substrate arranged, also can shorten luminous peak time simultaneously; (3) use gene engineering antigen, improved the specificity of reaction.
(biotin-avidin system BAS) has multistage signal amplification, and does not increase non-specific interference biotin-avidin system, and has highly sensitive, characteristics such as specificity good, stability is high, applicability is strong and experimental cost is low.
Chemiluminescence immune assay is a kind of elder generation and then effective method that detects hepatitis B surface antibody in conjunction with biotin-avidin system and immune magnetic particle solid phase separation system, helps lend some impetus to application and the development of chemiluminescence immunoassay technology in clinical examination, detection.
Summary of the invention
The present invention has solved the problems referred to above simultaneously, combine having a few of biotin-avidin immunity amplifying technique and magnetic particle immunity isolation technics and chemiluminescence immunoassay technology, developed a kind of hepatitis B surface antibody easy, quick, stable, that the range of linearity is wide and measured kit.
The purpose of this invention is to provide a kind of biotin-avidin immunity amplifying technique and immune magnetic particle technology magnetic particle chemiluminescent immunoassay kit in conjunction with the chemiluminscence immunoassay hepatitis B surface antibody.
Kit of the present invention comprises: a) mixed liquor of the magnetic particle of hepatitis B surface antigen bag quilt and biotin labeled hepatitis B surface antigen; B) streptavidin of horseradish peroxidase-labeled; C) hepatitis B surface antibody calibration object; D) chemical luminous substrate luminol that above-mentioned horseradish peroxidase acted on or different luminol; And e) concentrated cleaning solution; F) reaction tube.
A further object of the present invention provides a kind of method for preparing the mentioned reagent box.
The preparation method of kit of the present invention may further comprise the steps:
A) preparation hepatitis B surface antigen bag is by the mixed liquor of magnetic particle and biotin labeling hepatitis B surface antigen;
B) with the horseradish peroxidase-labeled streptavidin;
E) with hepatitis B surface antibody preparation calibration object;
D) the preparation chemical luminous substrate liquid that horseradish peroxidase acted on;
E) preparation concentrated cleaning solution;
F) the above-mentioned calibration object of packing, Avidin bag are by the mixed liquor of magnetic particle and biotin labeling antigen, enzyme labeling thing, chemical luminous substrate luminol or different luminol and reaction tube;
G) be assembled into the finished product kit.
The method according to this invention, in the step a) of the mixed liquor of antigen coated magnetic particle of described preparation and biotin labeling antigen:
Described mixed liquor is to be to mix at 1: 1 by the magnetic particle of hepatitis B surface antigen bag quilt and biotin labeled hepatitis B surface antigen with volume ratio, and the NBCS with 10～30%, the phosphate buffer of 0.1～1%Tween20 are formulated;
Described magnetic particle is that 1～10 μ m particle diameter, tri-iron tetroxide kernel, surface parcel have reactive group as amino (NH
2-), (polymkeric substance COOH), its working concentration are 1～10mg/mL to carboxyl;
Described antibody sandwich magnetic particle be by the glutaraldehyde two-step approach with the hepatitis B surface antigen bag by on magnetic particle;
With the magnetic particle of confining liquid sealing bag quilt, described confining liquid is that the pH value that contains 0.5%～2.0% bovine serum albumin(BSA), 10～20% NBCSs is 7.2 0.02mol/L phosphate buffer;
Described biotin labeling hepatitis B surface antigen realizes with antibody free lysine generation coupling reaction under the alkalescence condition by the biotin succinimide ester.
The method according to this invention in described step b) with the horseradish peroxidase-labeled streptavidin, adopts improvement sodium periodate method with the horseradish peroxidase-labeled streptavidin.
The method according to this invention, described chemical luminous substrate comprise A liquid and B liquid, wherein,
A liquid is the 0.2MpH8.7 boric acid-borate buffer solution that comprises 10mM luminol, 0.3mM 4-xenol, 0.05mM 4-iodobenzene boric acid, and its pH value is 8.0～10.0.
B liquid is the 0.2M pH7.2 phosphate buffer that comprises 3.5mM urea peroxide, 0.1%Tween20, and its pH value is 7.0～7.6.
In the method for mentioned reagent box produced according to the present invention, use transparent plastic or glass as the reaction tube material.
Concrete mentioned reagent box can comprise mixed liquor, enzyme labeling thing, calibration object, chemical luminous substrate and the lavation buffer solution etc. of antibody sandwich magnetic particle and biotin labeling antibody.Wherein, the raw material of described calibration object is a standard level, and purity is not less than 90%; Antibody sandwich forms mixed liquor with biotin labeled antibody on magnetic particle; The enzyme of mark streptavidin is a horseradish peroxidase; Chemical luminous substrate is a luminol; Lavation buffer solution is a phosphate buffer.
According to kit of the present invention, the magnetic particle of hepatitis B surface antigen bag quilt, the hepatitis B surface antibody in the testing sample combine with biotin labeled hepatitis B surface antigen, form " double antigens sandwich " composite structure, the streptavidin of enzyme labeling combines with this composite structure more afterwards, form " magnetic particle-antigen-antibody-antigen to be measured-biotin-avidin-enzyme " compound, the last also amplifying optical signals that produces with the chemical luminous substrate effect carries out highly sensitive, high special detection to hepatitis B surface antibody in the tubular type light-emitting appearance.The present invention adopts " the direct sandwich two-step approach of two antigens " reaction pattern, effectively utilized chemiluminescence in conjunction with magnetic particle and biotin-avidin immunity amplifying technique principle, hepatitis B surface antibody in detection by quantitative human serum, the plasma sample has been guaranteed the sensitivity that detects.The magnetic particle that uses has super paramagnetic, high dispersive, characteristics that surface area is big.The pre-treatment of sample is required low, sample pretreatment process is simple and reliable, can be fast, the high throughput testing gross sample, be convenient to operation and production.
The present invention's " hepatitis B surface antibody magnetic particle chemiluminescent immune analytic reagent kit " can detect the content of people's hepatitis B surface antibody exactly.Kit of the present invention (chemoluminescence method, biotin-avidin immunity amplifying technique combine with immune magnetic particle) detects fast, the measured value range of linearity is wide, calibration object is demarcated with national linear reference product, can detect the content of human serum hepatitis B surface antibody exactly, and every index all reaches the level of the similar kit of import.
Description of drawings
Fig. 1 is the correlation curve of kit of the present invention with the clinical blood sample measured value comparison of the external import HBsAb of renowned company chemical illuminating reagent.
Embodiment
Embodiment 1 preparation hepatitis B surface antibody magnetic particle chemiluminescent immune analytic reagent kit of the present invention
One, the hepatitis B surface antigen bag is by the preparation of the mixed liquor of magnetic particle and biotin labeling hepatitis B surface antigen
1, the preparation of the magnetic particle of hepatitis B surface antigen bag quilt
With particle diameter is that the magnetic particle of 1～10 μ m activates with glutaraldehyde, stirring at room, and mixing is after 3 hours, add magnetic field, leave standstill 30min, pour out supernatant, with the pH value is that 7.4 0.01mol/L phosphate buffer cleans three times, and suspends with this solution, and concentration is 50～100mg/mL; Add hepatitis B surface antigen 40～100 μ g in every milliliter of suspending liquid, after 4 ℃ stirring is spent the night down, add magnetic field, leave standstill 30min, pour out supernatant, the phosphate buffer (pH is 7.2) of using the 0.02mol/L that contains 0.2%～1.0% bovine serum albumin(BSA), 10～20% NBCSs was in room temperature sealing 3～4 hours; At last with the pH value be 7.4, the phosphate lavation buffer solution that contains NBCS, Tween 20 and biological preservative cleans 3～5 times, and is mixed with the working fluid of 1～10mg/mL with this solution.Magnetic particle is 2～8 ℃ of preservations.
2, the preparation of biotin labeled hepatitis B surface antigen
The biotin labeling hepatitis B surface antigen with the biotin succinimide ester under the alkalescence condition with antigen generation coupling reaction, PBS is fully dialysed, the biotin labeling thing dilutes with NBCS, adds Proclin300, preserves below-20 ℃.
3, the preparation of mixed liquor
Is to mix at 1: 1 the magnetic particle of hepatitis B surface antigen bag quilt and biotin labeled hepatitis B surface antigen with volume ratio, formulated with the phosphate buffer that contains 10～30% calf serums, 0.1～1%Tween 20.
Two, the preparation of the streptavidin of horseradish peroxidase-labeled
Adopt improvement sodium periodate method with the horseradish peroxidase-labeled streptavidin, concrete labeling process is as follows: dissolving 4.4mg HRP is in 1mL distilled water, add 0.4mL sodium periodate (50mmol/L) stirring at room 20min, through the 1mmol/L sodium-acetate buffer, pH 4.4 dialysis backs add the 8mg streptavidin, stir 2h, use 200mmol/L NaBH at last
4Reduce, after the dialysis of 0.02M PBS damping fluid, add equal-volume glycerine, preserve below-20 ℃.
Three, the preparation of hepatitis B surface antibody calibration object
With horse serum hepatitis B surface antibody is diluted to calibration object, concentration is respectively 0mIU/mL, 6mIU/mL, 20mIU/mL, 60mIU/mL, 200mIU/mL, 600mIU/mL, totally 6 bottles.
Four, the preparation of enzyme labeling thing
Preparation enzyme dilution contains Tris 12.12g in every 1000ml enzyme dilution earlier, BSA 5g, and glycerine 100ml, Proclin300 1ml adds distilled water to 900ml, transfers pH to 7.4 with hydrochloric acid, is settled to 1000ml with distilled water again, is made into the enzyme dilution.Adopting the square formation method to select the working concentration scope of enzyme labeling thing is 1: 1000～5000.
Five, preparation chemical luminous substrate liquid
The compound method of the chemical luminous substrate liquid of horseradish peroxidase used in the present invention (HRP):
A liquid: the borate buffering that contains pH8.0～10.0 of 10mM luminol, 0.3mM4-xenol, 0.05mM4-iodobenzene boric acid.
B liquid: pH7.0～7.6 phosphate buffers that contain 3.5mM urea peroxide, 0.1%Tween20.
Using method: A, B liquid bi-component reagent mix according to the use amount equal-volume before use.
Six, preparation lavation buffer solution
Lavation buffer solution is the phosphate buffer that contains 0.1～0.5% Tween-20,0.1% biological preservative, and the pH value is 7.4.Use 20 times of distilled water dilutings during use.
Seven, semi-manufacture and finished product are formed
The packing of above-mentioned steps products obtained therefrom is semi-manufacture.Extract three parts out and cross specificity, accuracy, sensitivity and stable assay approval through spirit and just can be assembled into hepatitis B surface antibody magnetic particle chemiluminescent immunoassay kit, be assembled into also need inspect by random samples behind the kit and just can dispatch from the factory after qualified.
The using method of embodiment 2 kits of the present invention
One, the preparation of sample
Adopting correct medical approaches to collect the patients serum is used for detecting.
Two, detection method
Before using this kit to experimentize, the Avidin of the mixed liquor of antigen coated magnetic particle of taking-up and biotin labeling antigen, calibration object/testing sample, enzyme labeling was placed 15～30 minutes in room temperature earlier, made them equilibrate to room temperature; Afterwards, constant temperature incubator or water-bath are transferred to 37 ℃; Again, be ready to suitable micro sample adding appliance and corresponding suction nozzle and check whether operate as normal of Chemiluminescence Apparatus.
Use kit of the present invention as follows according to the concrete operations step that the method for embodiment 2 experimentizes:
After round bottom polystyrene test tube numbering, in test tube, add 50 μ L blood serum samples or serial calibration object solution, the every pipe of calibration object adds 0mIU/mL, 6mIU/mL, 20mIU/mL, 60mIU/mL, each 50 μ L of 200mIU/mL, 600mIU/mL, the mixed liquor 50 μ L that add antigen coated magnetic particle and biotin labeling antigen again, 37 ℃ of oscillating reactions 30min.After cleansing solution cleaning three times, add the streptavidin 50 μ L of horseradish peroxidase-labeled, 37 ℃ of oscillating reactions 15min.Afterwards, test tube rack placed separate 5min on the magnetic separator, the separation vessel that reverses is then poured out supernatant, the test tube of reversing is placed on the filter paper blots, and bounces separation vessel to remove wall built-up liquid.Every pipe adds cleansing solution 500 μ L, and fully mixing places and separates 5min on the magnetic separator, pours out supernatant, the test tube that reverses is placed on the filter paper blots, and bounces separation vessel to remove wall built-up liquid, repeats 3 times.Each pipe adds chemical luminous substrate 200～400 μ L, and fully mixing places in the magnetic separator, treat that magnetic particle is enriched in the bottom after, 5min is placed in the dark place, then measures the luminous intensity (RLU) of each pipe on tubular type chemiluminescence measuring instrument in regular turn.Log value with calibration object concentration is a horizontal ordinate, and the Log value of RLU is drawn typical curve (double logarithmic curve) for ordinate, finds the concentration of the HBsAb of this serum on typical curve with each test serum RLU value.
The methodology calibrating of embodiment 3 kits of the present invention
According to manufacturing conventional in this area and vertification regulation the kit of preparation among the embodiment 1 is examined and determine, the result is as follows:
1, kit precision is measured
(1) calibration object precision experiment
The kit of preparation among the embodiment 1 is got three batches respectively carry out the precision experiment, every batch is extracted 5 kits.Anti--HBs calibration object 10 times with the kit measurement 80mIU/mL that extracted among the embodiment 1.Calculate the coefficient of variation of measuring concentration, the result shows that the coefficient of variation is between 3.0%～8.5%.
2, the kit accuracy is measured
Detect with national linear reference product, the measured value deviation of 8 concentration point is less than 10%, result such as table 1.
The national linear reference product of table 1 detect the result
| Indicate concentration (mIU/ml) | Measured value concentration (mIU/ml) | Deviation (%) |
| 160 | 162.75 | -1.72 |
| 106.7 | 110.18 | -3.26 |
| 71.1 | 74.53 | -4.82 |
| 47.4 | 50.53 | -6.6 |
| 31.6 | 31.86 | -0.82 |
| 21.1 | 20.1 | 4.74 |
| 14 | 13.49 | 3.64 |
| 9.4 | 8.87 | 5.64 |
3, kit sensitivity, specificity experiment
Do not see cross reaction with diseases such as HAV, HCV, rheumatoid disease, systemic loupus erythematosuses.Detect with national reference material, wherein 20 parts of national reference materials of feminine gender all detect feminine gender, coincidence rate 100% (20/20), and three parts of sensitivity reference serums detect result such as table 2.
Table 2 sensitivity country reference material testing result
| The sensitivity reference material | National standard | Testing result |
| 2mIU/ml | ± | ± |
| 5mIU/ml | ± | + |
| 10mIU/ml | + | + |
4, kit stability experiment
After the kit of embodiment 1 carried out 37 ℃ of 6 days accelerated tests, place the high, medium and low value quality-control product of the parallel detection of kit with 4 ℃, the result shows that 37 ℃ of quality controlled serum measured value deviations after 6 days are all less than 15%, embodiment 1 kit is carried out 2～8 ℃ of tracking tests of 8 months, and the result shows that every index meets clinical requirement fully.This kit was placed 8 monthly energy by national reference material standard in 6 days and 2～8 ℃ 37 ℃ of placements, had good stability, and met clinical needs fully.
Embodiment 4 kits of the present invention are with the clinical blood sample measured value comparison of external kit
Import with kit of the present invention and external renowned company is anti--and the HBs chemical luminescence reagent kit detects simultaneously to 512 parts clinical blood serum sample, its testing result is seen accompanying drawing 1, with the blood sample that the inventive method is measured anti--HBs result is a horizontal ordinate, the result who measures with the contrast agents box is that ordinate is done regretional analysis, dependent equation is: y=0.8413x+58.743, correlation coefficient r=0.9376.Learn result by statistics and show, the inventive method is good with the external clinical blood sample measured value of kit correlativity.
Claims (10)
1, a kind of hepatitis B surface antibody magnetic particle chemiluminescent immunoassay kit; it is characterized in that described kit comprises following component: the hepatitis B surface antigen bag by magnetic particle and the mixed liquor of biotin labeled hepatitis B surface antigen, the streptavidin of horseradish peroxidase-labeled, chemical luminous substrate liquid, concentrated cleaning solution and the reaction tube that hepatitis B surface antibody calibration object, above-mentioned horseradish peroxidase are acted on.
2, kit as claimed in claim 1 is characterized in that, the magnetic particle of described hepatitis B surface antigen bag quilt and the blending ratio of biotin labeled hepatitis B surface antigen are 1: 1.
3, kit as claimed in claim 1 is characterized in that, described chemical luminous substrate comprises A liquid and B liquid, wherein,
A liquid comprises 10mM luminol, 0.3mM 4-xenol, 0.05mM 4-iodobenzene boric acid, 0.2M pH8.7 boric acid-borate buffer solution, and its pH value is 8.0～10.0;
B liquid comprises 3.5mM urea peroxide, 0.1%Tween20,0.2M pH7.2 phosphate buffer, and its pH value is 7.0～7.6.
4, kit as claimed in claim 1 is characterized in that, the reaction tube material that described kit uses is transparent plastic or glass.
5, a kind of method for preparing the described kit of claim 1 is characterized in that may further comprise the steps:
1) preparation hepatitis B surface antigen bag is by the mixed liquor of magnetic particle and biotin labeling hepatitis B surface antigen;
2) with the horseradish peroxidase-labeled streptavidin;
3) with hepatitis B surface antibody polyclonal antibody preparation calibration object;
4) preparation chemical luminous substrate luminol or different luminol that horseradish peroxidase acted on;
5) concentrated cleaning solution
6) mixed liquor of the above-mentioned calibration object of packing, antigen coated magnetic particle and biotin labeling antigen, enzyme labeling thing, chemical luminous substrate luminol or different luminol, concentrated cleaning solution and reaction tube; And
7) be assembled into the finished product kit.
6, method as claimed in claim 5 is characterized in that, in the step 1) of the mixed liquor of antigen coated magnetic particle of described preparation and biotin labeling antigen:
Described magnetic particle is 1～10 μ m particle diameter, tri-iron tetroxide kernel, the surperficial polymkeric substance that has reactive group that wraps up, and its working concentration is 1～10mg/mL;
Described antibody sandwich magnetic particle be by the glutaraldehyde two-step approach with the hepatitis B e antibody bag by on magnetic particle, be that 7.2 0.02mol/L phosphate buffer is the magnetic particle that quilt is wrapped in the confining liquid sealing to contain 0.5%～2.0% bovine serum albumin(BSA), pH value;
Described biotin labeling hepatitis B surface antigen realizes with antibody lysine residue generation coupling reaction under the alkalescence condition by the biotin succinimide ester;
Described mixed liquor is to be to mix at 1: 1 by the magnetic particle of hepatitis B surface antigen bag quilt and biotin labeled hepatitis B surface antigen with volume ratio, and is formulated with the phosphate buffer that contains 10～30% NBCS, 0.1～1%Tween 20.
7, method as claimed in claim 6 is characterized in that, described reactive group is amino or carboxyl.
8, method as claimed in claim 5 is characterized in that, in described step 2 with the horseradish peroxidase-labeled streptavidin) in, adopt improvement sodium periodate method with the horseradish peroxidase-labeled streptavidin.
9, method as claimed in claim 5 is characterized in that, described chemical luminous substrate liquid comprises A liquid and B liquid, wherein,
A liquid comprises 10mM luminol, 0.3mM 4-xenol, 0.05mM 4-iodobenzene boric acid, 0.2M pH8.7 boric acid-borate buffer solution, and its pH value is 8.0～10.0;
B liquid comprises 3.5mM urea peroxide, 0.1%Tween20,0.2M pH7.2 phosphate buffer, and its pH value is 7.0～7.6.
10, method as claimed in claim 5 is characterized in that, uses transparent plastic or glass as the reaction tube material.
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