CN1453588A - Reagent for colloidal gold chromatographic analysis of SARS coronavirus antigen - Google Patents

Reagent for colloidal gold chromatographic analysis of SARS coronavirus antigen Download PDF

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CN1453588A
CN1453588A CN 03126894 CN03126894A CN1453588A CN 1453588 A CN1453588 A CN 1453588A CN 03126894 CN03126894 CN 03126894 CN 03126894 A CN03126894 A CN 03126894A CN 1453588 A CN1453588 A CN 1453588A
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antibody
sars coronavirus
coated
standby
reagent
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CN1159580C (en
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王继华
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WONDFO BIOTECH CO Ltd
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WONDFO BIOTECH CO Ltd
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Abstract

The present invention relates to one kind of reagent for colloidal gold chromatographic analysis of SARS coronavirus antigen. The reagent consists of coating film, absorbent paper, and glass fiber film coated with gold mark antibody adhered successively together. The present invention is used in colloidal gold chromatographic analysis of SARS coronavirus antigen in hospital, airport, custom, household, etc. Using it can obtain detection result in several minutes and this is favorable to prevent diffusion of epidemic situation.

Description

Colloidal gold chromatography detects the reagent of sars coronavirus antigen
Technical field
The present invention relates to measuring technology, colloidal gold chromatography detects the reagent of sars coronavirus antigen specifically.
Background technology
SARS (Severe Acute Respiratory Syndrome) (claims SARS (Severe Acute Respiratory Syndrome) again, Severe Acute RespiratorySyndrome, abbreviation SARS) wreaked havoc nearly five months in the whole world, April 16, the World Health Organization (WHO) announced that a kind of human past is the cause of disease of SARS from undiscovered novel coronavirus.Coronavirus (Corona Virus) be nineteen sixty-eight by discoveries such as Almeida, called after " coronavirus " is because of its form, this virus has the spinous process of the similar corona of shape on the coating under electron microscope.The coronavirus classification is extensive, but infected person and domestic birds and animals can cause the infective bronchitis of poultry, mouse hepatitis, porcine encephalomyelitis, feline infectious peritonitis etc.; The human corona virus can cause the infection of the upper respiratory tract, infant asthma and capillary bronchitis, acute gastroenteritis etc.
The researchers of countries in the world are studying the method for testing of rapid and precise sars coronavirus laboratory diagnosis.Yet after the standard reagent and sufficient method of testing that is used for virus and antibody test arranged, the diagnosis of SARS disease remained based on clinical and EPDML evidence.At present the laboratory testing method of sars coronavirus has: 1. molecular testing (being called for short the PCR method); 2. antibody test comprises ELISA method and IFA method at present; 3. cellular incubation.
1. molecular testing (PCR method)
The PCR method can be measured the genetic stew (comprising samples such as blood, ight soil, respiratory secretions or bodily tissue) of sars coronavirus in different samples.Primer is the main segment in the PCR method of testing, is announced on World Health Organization (WHO) website by the laboratory network of the World Health Organization (WHO).Developed the PCR testing tool that comprises primer, the positive and negative controller.
Generally speaking, existing P CR method of testing has extraordinary specificity, but lacks sensitivity.This test result that just means feminine gender can not be got rid of the existence that SARS virus is arranged among the patient.And the pollution owing to lacking the laboratory sample that the laboratory quality control causes can cause the appearance of false positive results.
Positive findings for the PCR test that has necessary Quality Control Procedure: the lab investigation of recommending to be used for sars coronavirus are to have extraordinary specificly, and positive findings means the existence of the genetic stew (being RNA) that sars coronavirus is arranged in sample.But and do not mean that the existence of live virus or exist a large amount of virus and enough infect other people.The negative findings of PCR test can not be got rid of the existence of SARS virus.With PCR method sars coronavirus is tested, owing to the reason the possibility of result appearance of following several respects is negative:
1. patient is not infected by sars coronavirus; Case is to be infected by other pathogen (virus, bacterium and fungi) to cause, or owing to noninfective reason causes.
2. test result is incorrect (false negative).Present method of testing needs further to improve to improve its sensitivity.
3. sample is not to collect when having virus or genetic stew to exist.Virus and genetic stew might only exist only in the short period, depended on the kind of the sample that is used to test.
The PCR method only is used for the detection of viral nucleic acid, requires to possess pcr amplification instrument and gel electrophoresis equipment, and experimental period is grown (more than two hours), needs professional and technical personnel's operation and judges testing result.
2. antibody test
Behind the coronavirus infection human body, body will produce corresponding antibodies (people's anti-coronavirus antibody), in order to resist the propagation of virus.When human body by the SARS virus infections and after producing antibody, promptly use patient's serum (antibody) and detectable to react, the explanation experimenter who is positive is infected by the virus, and has produced antibody (immune response).But for the early stage patient who does not also produce antibody of morbidity, negative findings can not illustrate that the experimenter is not infected by the virus.
Detect the level of patient's internal antibody IgM and IgG specifically, the patient who has these two kinds of antibody to produce can be defined as SARS the infected.Still need follow the tracks of detection to negative findings, still not detect antibody after latent period then can get rid of ill possibility if surpass SARS.
1. ELISA (enzyme linked immunological absorption reaction)
The ELISA method is according to antigen and antibody specific immune response principle design, adopts indirect method or dual-antigen sandwich method to detect SARS coronary virus resistant IgG antibody and IgM in the serum of SARS case, and reliable positive findings appears in big after disease begins 10 days.These method characteristics are high specificities, be quick on the draw, the recall rate height, are the main method of The World Health Organization's pathogenic autoantibody clinical detection of recommending.
Indirect method is mainly used in the antibody composition that detects in the body fluid.This method is the surface that specific antigen is coated on solid phase carrier, form solid phase antigen, add sample to be checked, corresponding antibodies wherein is attached on the antigen of solid phase carrier, the antiantibody that adds enzyme labeling again, form Ag-Ab-enzyme labeling antiantibody compound, add the substrate solution colour developing at last.Substrate for enzymatic activity on the solid phase carrier becomes coloured product, predicts the content of antibody in the sample by colorimetric.The advantage of indirect method is as long as the conversion envelope antigen just can utilize same enzyme labeling antiantibody to set up the method that detects corresponding antibodies.This method test key of success is the purity of envelope antigen.Pay special attention to remove the impurity that deenergizes with general health human serum generation immunological response, can not contain the material that can must resist people Ig (s) reaction in the envelope antigen with enzyme labeling.The non-specific IgG of high concentration that another disturbing factor is in the normal serum to be contained in the indirect method.Because the adsorbability of IgG is very strong, non-specific IgG can directly be adsorbed on the solid phase carrier, also can be adsorbed on the surface of envelope antigen sometimes; So in indirect method experiment, generally will seal the remaining space on the solid phase carrier after antigen coated with irrelevant protein.The serum sample that detects generally can not directly use, and needs to add with envelope antigen after dilution to react, with the IgG that the reduces high concentration interference to experimental result again.
The reaction pattern and the double antibody sandwich method of dual-antigen sandwich method are similar.Specific antigen is combined on certain solid phase carrier, tracer-labelling on specific antigen, is added zymolyte, after enzyme-to-substrate reacts, the substrate colour developing, qualitative or detect the amount of antibody in the sample to be checked quantitatively according to the depth of substrate colour developing.The difference that it and indirect method are measured antibody is to replace the enzyme labeling antiantibody with enzyme-labelled antigen, and its susceptibility and specificity all are higher than indirect method.The key of this law is that the preparation of enzyme-labelled antigen should seek suitable labeling method according to the difference of antigenic structure.
The ELISA method requires antigen purity height, specificity good, otherwise nonspecific reaction can appear, running program is complicated, need cyclic washing, if washing times easily causes false positive and false negative inadequately or too much, and easily causing the operator to be injured and environmental pollution, experimental period is longer, needs to draw testing result more than two hours.This method must possess microplate reader and wash the plate machine, this basic unit laboratory and small-sized outpatient service difficulty reach, and test is subjected to condition restriction such as temperature, makes troubles to detection.
2. IFA (fluorescence immunoassay method of inspection)
The IFA method is equally according to antigen and antibody specific immune response principle design, and the cell fixation of infective virus on slide, is dripped patient's to be detected serum in top, have among the viral patients serum and contain corresponding antibodies, be incorporated on the slide, add fluorescent reagent, can demonstrate fluorescence.But in fluorescence immunoassay microscopically Direct observation testing result.IFA is used for the mensuration of IgG antibody of the serum of SARS case, and positive findings appears in big after disease begins 10 days.This method of testing also can be used for the mensuration of IgG antibody.This also is a kind of assay method of classics, need measure by means of the fluorescence immunoassay microscope.
The IFA method is highly sensitive, but must possess fluorescence immunoassay microscope and dark room conditions, must be equipped with suitable optical filter for obtaining the good fluorescence observing effect, experimental period needs two hours, the result is detected on the fluorescence immunoassay microscopically and carries out, and must judge testing result by experienced professional and technical personnel.
The infection of sars coronavirus before showing, positive antibody test result was once arranged.Seroconversion from the feminine gender to the positive took place from the acute stage to the convalescence, or antigen titration increased by four times, showing has infection in the recent period.
Negative antibody test result: behind 21 days of disease generation, be not checked through antibody, show the infection that is not subjected to sars coronavirus.
3. cellular incubation
From the virus in the sample of SARS case (for example respiratory secretions, blood or ight soil), also can measure by inoculating cell cultivation and virus multiplication.In case be separated to virus, will have done further and differentiate by methods such as electron microscopic observation or serological reactions whether to confirm be SARS virus.Cellular incubation is the very harsh test of condition, but (except animal testing) only shown the existence that live virus is arranged at present.The positive cells cultivation results shows the existence that sars coronavirus alive is arranged in the sample of being tested.Negative cellular incubation result can not get rid of exist (the seeing negative PCR result) of sars coronavirus.
Cellular incubation is a kind of aseptic technique, requirement possesses very high experimental work environment and condition (between sterile working, superclean bench, operation room etc.), need a large amount of instrument and equipments (incubator, hydro-extractor, microscope, hydro-extractor etc.), need a large amount of Specialty Experiment equipment (multiple cultivation vessel), the experimental technique complexity, needs experienced professional and technical personnel operate, and experimental period very long (needing the several months), is not suitable for the fast detecting of disease.
Summary of the invention
The objective of the invention is to problem at the prior art existence, provide a kind of colloidal gold chromatography to detect the reagent of sars coronavirus antigen, can be applicable to places such as hospital, airport, customs, family, can be in a few minutes judged result, thereby prevent the epidemic situation diffusion early.
The reagent of colloidal gold chromatography detection sars coronavirus antigen of the present invention is pasted successively by the glass fibre membrane of coated film, thieving paper, coated with gold labeling antibody and is constituted;
Described coated film prepares by following method: being cushioned liquid dilution coated antibody to concentration with bag is 5~20 μ g/ml, sprays or is coated on the coated film, after drying, soaks 60 minutes in 37 ℃ of confining liquids, takes out rearmounted 37 ℃ of oven dry 2 hours, and envelope is standby;
Described coated antibody is sars coronavirus monoclonal antibody or sars coronavirus polyclonal antibody or two kinds and two or more sars coronavirus monoclonal antibody.
Prescription and preparation method that described bag is cushioned liquid and confining liquid are the general technology in this area.
Described thieving paper is common commercially available.
Described golden labeling antibody obtains by following method: regulate collaurum pH value to 7.5 with 0.1M sal tartari, add the sars coronavirus monoclonal antibody by 9~11 μ g/ml collaurums, leave standstill behind the mixing, centrifugal treating, supernatant discarded, to precipitate with the washing of mark cleansing solution, the golden labeling antibody preservation liquid that will precipitate with 1/10th initial collaurum volumes dissolves, put 4 ℃ standby.
The glass fibre membrane of described coated with gold labeling antibody can be coated in golden labeling antibody on the glass fibre membrane with universal method, also can obtain by following method: the golden labeling antibody that will prepare is layered on the glass fibre membrane equably, 4~8 square centimeters of every ml soln shops, freeze drying, envelope, put 4 ℃ standby.
The principle of work of reagent of the present invention is as follows:
The colloidal gold immunochromatographimethod technology is a kind of rapidly easy, immunological detection method fast of development in recent years, this reagent adopts the colloidal gold immunochromatographimethod technology, select for use a kind of sars coronavirus monoclonal antibody (or sars coronavirus polyclonal antibody, two or more sars coronavirus monoclonal antibody) as solid formation, utilize the double antibody sandwich method principle to detect in the serum whether contain sars coronavirus antigen.When containing sars coronavirus antigen in the sample to be checked, antigen elder generation and golden labelled antibody combination, because chromatography action-reaction compound moves forward along coated film, when running into envelope antigen, form antibody-antigen-Jin labelled antibody compound and be enriched in bag by on the line, form the red precipitate line, positive result, thereby quick diagnosis SARS.
The present invention compared with prior art has following advantage:
1, diagnosis is quick, can finish detection in 10 minutes.
2, do not need any instrument and equipment.
3, easy and simple to handle, do not need to operate by the professional.
Embodiment
Embodiment 1
Present embodiment adopts a kind of sars coronavirus monoclonal antibody as solid formation, utilizes the double antibody sandwich method principle to detect in the serum whether contain sars coronavirus antigen.The manufacture method of present embodiment is as follows: a, Antibody Preparation adopt a kind of sars coronavirus monoclonal antibody.The preparation of b, antibody membrane
1, bag is cushioned the preparation of liquid: 0.05M pH9.6 sulfate damping fluid for bag by solution, 0.22 μ membrane filtration mistake, put 4 ℃ standby, one week of effect phase.
2, the preparation of confining liquid: the preparation 0.01M pH7.0 phosphate buffer (PBS), 0.22 μ membrane filtration mistake, put 4 ℃ standby, one week of effect phase.Preparation sealing working fluid: 2%BSA, 2% skimmed milk, 0.01M pH7.0PBS, 0.22 μ membrane filtration mistake, put 4 ℃ standby, three days effect phases.
3, coated film preparation: debugging Membrane jetter, spouting liquid are 20 microlitres/35 centimetre, are cushioned liquid dilution coated antibody with bag, and concentration is 8 μ g/ml, the machine line, and line-to-line should be careful even every 5mm, and room temperature was dried 20 minutes.37 ℃ in confining liquid soaks 60 minutes, take out rearmounted 37 ℃ of oven dry and handled two hours, envelope is standby.The preparation of c, collaurum, colloid gold label antibody
1, the preparation of gold chloride: with distilled water dissolved chlorine auric acid, be made into 1% solution, put 4 ℃ standby, three days effect phases.1000ml 1% chlorauric acid solution prescription: 10g gold chloride; The 1000ml distilled water.
2, the preparation of trisodium citrate: dissolve trisodium citrate with distilled water, be made into 1% solution, put 4 ℃ standby, three days effect phases.1000ml 1% citric acid three sodium solution prescription: 10g trisodium citrate; The 1000ml distilled water.
3, the preparation of 0.1M sal tartari: with distilled water preparation, 0.22 μ membrane filtration mistake, put 4 ℃ standby, one week of effect phase.1000ml 0.1M solution of potassium carbonate prescription: 13.8g sal tartari; The 1000ml distilled water.
4, the preparation of 3%PEG-20000: with distilled water preparation, 0.22 μ membrane filtration mistake, put 4 ℃ standby, one week of effect phase.1000ml 3%PEG solution formula: 30g PEG-20000; The 1000ml distilled water.
5, the preparation of mark cleansing solution: 2%BSA, 0.01M pH7.0 PBS solution, 0.22 μ membrane filtration mistake, put 4 ℃ standby, two weeks of effect phase.1000ml mark cleansing solution prescription: 20gBSA; 1000ml0.01M pH7.0PBS solution.
6, golden labeling antibody is preserved the preparation of liquid: 1%BSA, 0.5% skimmed milk, 5/0,000 NaN3,0.1%Tween-20,0.01M pH7.0 PBS solution, 0.22 μ membrane filtration mistake, put 4 ℃ standby, 15 days effect phases.1000ml gold labeling antibody is preserved formula of liquid: 10g BSA; The 5g skimmed milk; 0.5g NaN3; 1ml Tween-20; 1000ml0.01M pH7.0 PBS solution.
7, firing of collaurum: 1% gold chloride is diluted to 0.01% with distilled water, put electric furnace and boil, add 4 milliliter of 1% trisodium citrate, continue to boil by per 100 milliliter of 0.01% gold chloride, be shiny red up to liquid and promptly stop heating, supply dehydration after being cooled to room temperature.Outward appearance should be pure, and is bright, do not have precipitation and floating thing.
8, the preparation of golden labeling antibody: transfer collaurum pH value to 7.5 with 0.1M sal tartari, add the sars coronavirus monoclonal antibody by 10 micrograms antibody/milliliter collaurum, mixing left standstill 30 minutes, centrifugal 30 minutes of 13000rpm, abandon supernatant, precipitation mark cleansing solution washed twice, last supernatant discarded will precipitate with the golden labeling antibody of 1/10th initial collaurum volumes and preserve the liquid dissolving, put 4 ℃ standby, one week of effect phase.The freeze-drying of d, golden labeling antibody
The collaurum that mark is good is layered on the glass fibre membrane equably, 6 square centimeters of every ml soln shops, freeze drying, envelope, put 4 ℃ standby.The production of e, test paper plate
1, cutting of golden labeling antibody glass fibre membrane: join the width that experiment is determined according to dripping, golden labeling antibody glass fibre membrane is cut, put between drying shed standby.
2, cutting of thieving paper: with trimmer thieving paper is cut into 35 centimeter length, 4.2 centimetres wide, puts between drying shed standby.
3, cutting of glass fibre membrane: glass fibre membrane is cut into long 35 centimetres, and wide 2 centimetres is rectangular, puts between drying shed standby.
4, the stickup of big plate: on request coated film, thieving paper, freeze-drying gold labeling antibody, glass fibre membrane are sticked on the plastic bottom board, form big plate.Composing room's temperature is controlled at 25 ℃, humidity 20%-30%.F, slitting
With cutting cutter big plate is cut into single person-portion, 2.5 millimeters of everyone part width, sampling observation at random, sensitivity can detect indoor Quality Control, and band colour developing degree reaches one "+" number, and specific band can pass through the Panel of the Ministry of Public Health nothing but.G, assembling, packing
The test strips and one that 50 person-portions have been cut the drying prescription of being responsible for a task until it is completed is encapsulated in the aluminium foil polybag, puts into kit, a instructions of every box.Keep in Dark Place in 4-25 ℃, must not be frozen.
Detect the 150 suspicious patients' of routine SARS serum sample, (PCR) compares with the molecular testing method, and the present invention is carried out specificity and sensitivity Detection (seeing Table 1).Specificity of the present invention reaches 92% (92/100), and sensitivity reaches 90% (45/50).
150 parts of suspicious patients serum's samples of SARS of table 1 sars coronavirus Detection of antigen result
Control group (PCR) The negative number of number positive Experimental group (the present invention) Add up to 50 100 150
Number positive 45 8 51 Negative several 5 92 97
Add up to
Embodiment 2
Present embodiment adopts two or more sars coronavirus monoclonal antibody as solid formation, utilizes the double antibody sandwich method principle to detect in the serum whether contain sars coronavirus antigen.The manufacture method of present embodiment is substantially the same manner as Example 1, and difference is as follows:
1, adopt two or more sars coronavirus monoclonal antibody as coated antibody.
2, the preparation of coated film: the debugging Membrane jetter, spouting liquid is 20 microlitres/35 centimetre, is cushioned liquid dilution coated antibody with bag, concentration is 5 μ g/ml, the machine line, line-to-line should be careful even every 5mm, and room temperature was dried 20 minutes.37 ℃ in confining liquid soaks 60 minutes, take out rearmounted 37 ℃ of oven dry and handled two hours, envelope is standby.
3, the preparation of golden labeling antibody: transfer collaurum pH value to 7.5 with 0.1M sal tartari, add the sars coronavirus monoclonal antibody by 11 micrograms antibody/milliliter collaurum, mixing left standstill 30 minutes, centrifugal 30 minutes of 13000rpm, abandon supernatant, precipitation mark cleansing solution washed twice, last supernatant discarded will precipitate with the golden labeling antibody of 1/10th initial collaurum volumes and preserve the liquid dissolving, put 4 ℃ standby, one week of effect phase.
4, the freeze-drying of golden labeling antibody: the collaurum that mark is good is layered on the glass fibre membrane equably, 4 square centimeters of every ml soln shops, freeze drying, envelope, put 4 ℃ standby.
Embodiment 3
Present embodiment adopts the sars coronavirus polyclonal antibody as solid formation, utilizes the double antibody sandwich method principle to detect in the serum whether contain sars coronavirus antigen.The manufacture method of present embodiment is substantially the same manner as Example 1, and difference is as follows:
1, adopt the sars coronavirus polyclonal antibody as coated antibody.
2, the preparation of coated film: the debugging Membrane jetter, spouting liquid is 20 microlitres/35 centimetre, is cushioned liquid dilution coated antibody with bag, concentration is 20 μ g/ml, the machine line, line-to-line should be careful even every 5mm, and room temperature was dried 20 minutes.37 ℃ in confining liquid soaks 60 minutes, take out rearmounted 37 ℃ of oven dry and handled two hours, envelope is standby.
3, the preparation of golden labeling antibody: transfer collaurum pH value to 7.5 with 0.1M sal tartari, add the sars coronavirus monoclonal antibody by 9 micrograms antibody/milliliter collaurum, mixing left standstill 30 minutes, centrifugal 30 minutes of 13000rpm, abandon supernatant, precipitation mark cleansing solution washed twice, last supernatant discarded will precipitate with the golden labeling antibody of 1/10th initial collaurum volumes and preserve the liquid dissolving, put 4 ℃ standby, one week of effect phase.
4, the freeze-drying of golden labeling antibody: the collaurum that mark is good is layered on the glass fibre membrane equably, 8 square centimeters of every ml soln shops, freeze drying, envelope, put 4 ℃ standby.

Claims (5)

1, a kind of colloidal gold chromatography detects the reagent of sars coronavirus antigen, it is characterized in that being pasted successively by the glass fibre membrane of coated film, thieving paper, coated with gold labeling antibody constituting; Described coated film prepares by following method: being cushioned liquid dilution coated antibody to concentration with bag is 5~20 μ g/ml, spray or be coated on the coated film, after drying, in 37 ℃ of confining liquids, soaked 60 minutes, take out rearmounted 37 ℃ of oven dry 2 hours, envelope is standby; Described coated antibody is sars coronavirus monoclonal antibody or sars coronavirus polyclonal antibody or two kinds and two or more sars coronavirus monoclonal antibody; Described golden labeling antibody obtains by following method: regulate collaurum pH value to 7.5 with 0.1M sal tartari, add the sars coronavirus monoclonal antibody by 9~11 μ g/ml collaurums, leave standstill behind the mixing, centrifugal treating, supernatant discarded, to precipitate with the washing of mark cleansing solution, the golden labeling antibody preservation liquid that will precipitate with 1/10th initial collaurum volumes dissolves, put 4 ℃ standby.
2, colloidal gold chromatography according to claim 1 detects the reagent of sars coronavirus antigen, the glass fibre membrane that it is characterized in that described coated with gold labeling antibody obtains by following method: the golden labeling antibody that will prepare is layered on the glass fibre membrane equably, 4~8 square centimeters of every ml soln shops, freeze drying, envelope, put 4 ℃ standby.
3, colloidal gold chromatography according to claim 1 and 2 detects the reagent of sars coronavirus antigen, it is characterized in that described coated film prepares by following method: being cushioned liquid dilution coated antibody to concentration with bag is 8 μ g/ml, spray or be coated on the coated film, after drying, in 37 ℃ of confining liquids, soaked 60 minutes, take out rearmounted 37 ℃ of oven dry 2 hours, envelope is standby.
4, colloidal gold chromatography according to claim 1 and 2 detects the reagent of sars coronavirus antigen, it is characterized in that described golden labeling antibody obtains by following method: regulate collaurum pH value to 7.5 with 0.1M sal tartari, add the sars coronavirus monoclonal antibody by 10 μ g/ml collaurums, leave standstill behind the mixing, centrifugal treating, supernatant discarded will precipitate with the washing of mark cleansing solution, to precipitate with the golden labeling antibody of 1/10th initial collaurum volumes and preserve the liquid dissolving, put 4 ℃ standby.
5, colloidal gold chromatography according to claim 1 and 2 detects the reagent of sars coronavirus antigen, it is characterized in that described coated film prepares by following method: being cushioned liquid dilution coated antibody to concentration with bag is 8 μ g/ml, spray or be coated on the coated film, after drying, in 37 ℃ of confining liquids, soaked 60 minutes, take out rearmounted 37 ℃ of oven dry 2 hours, envelope is standby; Described golden labeling antibody obtains by following method: regulate collaurum pH value to 7.5 with 0.1M sal tartari, add the sars coronavirus monoclonal antibody by 10 μ g/ml collaurums, leave standstill behind the mixing, centrifugal treating, supernatant discarded, to precipitate with the washing of mark cleansing solution, the golden labeling antibody preservation liquid that will precipitate with 1/10th initial collaurum volumes dissolves, put 4 ℃ standby.
CNB031268943A 2003-06-18 2003-06-18 Reagent for colloidal gold chromatographic analysis of SARS coronavirus antigen Expired - Fee Related CN1159580C (en)

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