CN1424585A - Quick enzyme immuno-determining method and box for food borne parasites - Google Patents
Quick enzyme immuno-determining method and box for food borne parasites Download PDFInfo
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- CN1424585A CN1424585A CN 03113564 CN03113564A CN1424585A CN 1424585 A CN1424585 A CN 1424585A CN 03113564 CN03113564 CN 03113564 CN 03113564 A CN03113564 A CN 03113564A CN 1424585 A CN1424585 A CN 1424585A
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Abstract
A high-speed enzyme immunoassay method for the parasitosis caused by food and its multi-hole detecting box are disclosed. Its reagent series is composed of antigen coated plate, enzyme compound liquid, lotion, substrate, color developer, sample diluent, stopping agent, postive reference and negative reference. Its detecting process includes such steps as diluting sample, adding sample, enzyme reacting and colouring reacting. Its advantages are high speed, easy operation and high reliability and effect.
Description
Technical field the present invention relates to the quick enzyme immunity detection method of a kind of food-borne parasitic disease and detects box.
Background technology is well-known, and food-borne parasitic disease (FOOD SOURCE PARASITEDISEASES) is one group can mainly comprise pork measles, lung fluke, liver fluke, four kinds of parasitic diseases of trichina by food-borne common parasitic disease.They can infect human body and domestic animal, and the grievous injury human health also causes the economic loss of animal husbandry.Wherein, pork measles can cause epilepsy, paralysis, blind; Lung fluke can cause pulmonary cyst, pulmonary abscess; Liver fluke can cause infection of biliary tract, gall stone, cirrhosis; Trichina then can cause severe anaphylactic reaction and heart damage.Food source property parasite is popular extensively, only just reaches more than 3,000 ten thousand at the Chinese Guangdong province popular district of liver fluke population.Even more noteworthy, the onset of food-borne parasitic disease concealment, atypical symptom, easily and other disease obscure, therefore the reliable and practical detection method of present in addition clinical shortage, is usually failed to pinpoint a disease in diagnosis mistaken diagnosis and is affected adversely the state of an illness.
At present, the detection method of food-borne parasitic disease has two big classes: a class is a traditional detection method, promptly search pathogen, because these four kinds of parasite multiparasitizations are in deep tissue, as internal organs such as brain, eyeball, liver biliary tract, lung, skeletal muscle, cardiac muscles, to gather polypide by surgical means, not only damage very greatly but also often polypide distribute to be dispersed in and often be difficult to seize.Trichina, pork measles parasite elimination ovum not in addition, seldom even ovulation of lung fluke ovulation; Though liver fluke can ovulate with ight soil, oophyte is long-pending small and ovulate irregularly, is difficult to detect, and Chang Yong fecal smear method susceptibility generally is lower than 50% clinically.Therefore, look into the effective means that worm's ovum or polypide are not this group parasitic diseases of diagnosis; An other class is exactly the conventional enzyme immunoassay that occurs in recent years etc., but this class methods ubiquity is following not enough: 1, the running time is long: reaching more than 2-4 hour time, be difficult to adapt to the quick diagnosis needs of on-the-spot and clinical outpatient service more; 2, reagent operation is loaded down with trivial details: to prepare as enzyme conjugates and substrate solution temporarily, and the antigen bag quilt that need spend the night, sample diluent liquid and stop liquid and all need the user to prepare has voluntarily been brought inconvenience to the user in application; 3, instability: the reagent stability that is provided is not good, in case preparation must use up in 1 week even in 1 day, causes economic loss, has influenced the reliability that detects yet; 4, supporting imperfect: so far, do not see four kinds of unified food source property parasite reagent set boxes of provider's method simultaneously as yet on market, all kits all can only be measured a kind of parasitic disease.At present, seen supplying unit supply kind is uneven complete, only supply wherein one or both, only provide the cysticercus detectable as Chinese somewhere a company, another research institute then only provides the trichina detectable with the area.The difference of producer, the difference of method, the difference of performance not only influences operability, more has influence on result's consistance and validity, causes mistaken diagnosis easily or fails to pinpoint a disease in diagnosis, and becomes a big hidden danger in the food-borne parasitic disease control; 5 potential safety hazards: the normal OPD with teratogenesis effect that uses of conventional enzyme immunization method is as developer, and with the sulfuric acid of severe corrosive as stop buffer, to operating personnel and environment structure threat.In sum, existing detection technique can not effectively satisfy the diagnosis needs of food-borne parasitic disease, is unfavorable for the diagnoses and treatment and the prevention and control of disease.
Summary of the invention the object of the present invention is to provide quick enzyme immunity detection method of food-borne parasitic disease and and the matched complete detection box of this method.
In order to achieve the above object, the invention provides following technical scheme: the quick enzyme immunity detection method of a kind of food-borne parasitic disease, be used for pork measles, lung fluke, the enzyme immune detection of liver fluke and trichina, the enzyme immune detection of this method comprises antigen coated microplate A with reagent, enzyme conjugates liquid B, cleansing solution C, substrate D, developer E, sample diluting liquid F, terminator G, positive reference substance H and negative control product I, testing process uses porous to detect box, detailed process comprises: the dilution of (1) sample: with sample dilution F serum to be checked was diluted in 1: 50 by volume, the yin and yang attribute control serum is dilution synchronously in proportion, (2) application of sample reaction then: every Kong Jiayi dilute serum 100ul, establish feminine gender simultaneously, each hole of the positive and blank, the blank hole adds 100ul sample diluting liquid F, 37 ℃ left standstill 10 minutes, get rid of, every hole adds one of cleansing solution C, immediately repeatedly with the distilled water washing, get rid of, pat dry, (3) add enzyme reaction again: add two of enzyme conjugates B, 37 ℃ were reacted 10 minutes, get rid of, add one of cleansing solution C, use the distilled water cyclic washing immediately repeatedly, get rid of, pat dry, (4) chromogenic reaction at last: add each one of substrate D and developer E, mixing, placed 2-3 minute under the room temperature, when treating that negative control hole is little and being blueness, add mixing of terminator G, cessation reaction, visual inspection is shallower than that to be bordering on negative control negative, be and obviously be deeper than the blue positive of negative control, judge with instrument: return to zero with blank and read the O.D value, treat that verify O.D value is positive greater than 2.1 times of persons of negative control, when negative control O.D value is lower than 0.07, calculate by 0.07 in 630nm.
In concrete testing process, must be noted that following problem: 1, kit is preserved down at 2-8 ℃, and elder generation's balance is used to room temperature when taking out at every turn.Drop bottle at every turn with after lid must be tightened, can't use with between each bottle cap; When 2, washing distilled water is filled it up with in the hole, parked for 30 seconds at every turn, get rid of liquid in the hole, other water sources such as forbidding tap water.
The present invention's technical scheme preferably can be: antigen coated microplate A adopts Fast Packet by technology, envelope antigen on the general purpose polystyrene microwell plate in advance, coating buffer is that the carbonate buffer solution that contains an amount of antigen (is made up of sodium bicarbonate and sodium carbonate, PH9.60,0.05M), static 12 hours of room temperature was with 1% bovine serum albumin(BSA) sealing 2 hours, vacuum freezedrying, vacuum bagging seals.Adopt Fast Packet can be increased work efficiency and improve the accuracy rate of detection by technology.
The present invention's technical scheme preferably also can be: enzyme conjugates B liquid phosphate buffer (is made up of sodium dihydrogen phosphate and sodium hydrogen phosphate, PH7.0,0.05M), interpolation stabilizing agent molecular weight Mr is 20000 polyglycol weight 0.5%-5%, add accelerator molecular weight Mr simultaneously and be 6000 polyglycol weight 0.5%-5%, add ten thousand/thimerosal and four kinds of anti-immunoglobulin G while horseradish peroxidase-labeled bonds that parasitic disease is unified.
In implementing process of the present invention, other composition or collocation method of reagent that detects usefulness is as follows: cleansing solution C: (be made up of sodium dihydrogen phosphate and sodium hydrogen phosphate, PH7.0 0.05M), adds ten thousand/thimerosal to adopt the phosphate buffer of 0.5% tween; Substrate solution D and developer E: substrate solution is a superoxol, and developer is harmless TMB solution; Sample dilution F: (be made up of sodium dihydrogen phosphate and sodium hydrogen phosphate, PH7.0 0.1M), contains tween 0.5% to phosphate buffer; Stop buffer G:1% lauryl sodium sulfate (SDS); Positive control H and negative control I: positive control is with four kinds of positive patients serum inactivation treatment, mixes, and adds the packing of ten thousand/thimerosal, become can be general the mixing positive control; Negative control is after some normal human serums mix, and adds the packing of ten thousand/thimerosal and preserves.Above component has comprised detection needed whole reagent (except the distilled water), and wherein the equal adjusted of liquid component need not be diluted and can directly use in the working concentration and the plastic dropping bottle of packing into; Plastic tube with cover is put in the negative and positive contrast, can directly use; Bag also can directly use after being sealed off by plate.
The present invention's technical scheme preferably can also be: it is a kind of kit of four unificationizations that can fasten that porous detects box, be divided into two districts in the box, place multiple parasite antigen and wrap in advance by plate in a district, be total to polylith immediate vicinity of fluid reagent bottle, another district is provided with holder of a perforate paper or the multiple general liquid reagent bottle of the vertical parallel placement of plastic base, there is opening the kit top, and a hole is respectively arranged on the inside cover, inserts feminine gender and positive control serum respectively.In actual the use, be used to coordinate four kinds of antigen coated conditions and make other liquid component that comprises enzyme conjugates can become the common reagent that four kinds of parasites detections all can directly be used, therefore only can satisfy the detection requirement of four kinds of parasitic diseases once the cover liquid reagent; Be divided into two districts in the box, place four kinds of parasite antigens and wrap in advance by plate in a district, totally four immediate vicinity of fluid reagent bottles, the arrangement of standing parallel to each other; Another district is provided with the holder of a perforate paper or 6 kinds of general liquid reagent bottles of the vertical parallel placement of plastic base.The kit opening is located in the above, designs a hole on the inside cover respectively, inserts feminine gender and positive control serum respectively.
Compared with prior art, the present invention has following tangible advantage: 1, improved reaction velocity, the entire reaction time is no more than 30 minutes.Improved more than 4 times than conventional enzyme immunization method; 2, convenient and practical: kit not only has been equipped with and has detected necessary reagent component, and is model of instant use once to turn it on, and the user needn't prepare or dilute separately, makes that operation is simple.Can also can directly judge by naked eyes that not only limited scene generaI investigation and the outpatient service of the condition that is suitable for especially detects with the quantitative judged result of instrument; 3, improved the reliability of four kinds of disease detection: owing to unified method of operating and liquid reagent, make the user to operate at one time with under the method, make the result have the consistance and the comparability of height; 4, safety and environmental protection; Adopted harmless TMB reagent, harm and the pollution of having avoided conventional method to bring.Stop buffer adopts non-corrosive SDS, has eliminated the potential safety hazard because of using the sulfuric acid stop buffer to be brought; 5, detect effective: (1) susceptibility: detect through 453 parts of the liver rot human serums made a definite diagnosis with this quick kit, detect positive case 417 examples, susceptibility reaches 92.10% (417/453); (2) specificity: detect the healthy human serum in the 356 non-popular districts of example with this quick kit, detect negative sample 350 examples, specificity reaches 98.30% (350/356); (3) authenticity (youden index, Youder ' s Index): youden index is 0.904, has shown the authenticity that detects the box height; (4) repeatability: 50 parts of 56 parts of same operator's duplicate detection positive serums and negative serums, repeatability reaches 96.20%; (5) stability: will detect box and be placed under 4 ℃, dynamic observe six months, front and back are there was no significant difference as a result.
Be detection box drawing explanation of the present invention below the description of drawings:
Fig. 1 is the structural representation that detects box;
Among the figure: 1 is the coated plates of four kinds of parasite antigens, the 2nd, and control serum, the 3rd, place examination The base of agent bottle, the 4th, reagent bottle; With reference to Fig. 1, it is a kind of can the fastening that porous detects box The kit of four unificationizations is divided into two districts in the box, district's multiple parasite antigen of placement is pre-Coated plate is total to polylith immediate vicinity of fluid reagent bottle, and another district arranges the holder of a perforate paper or plastic base Hold in the palm the presser for liquid reagent bottle of vertical parallel placement many general, there are opening, inboard in the kit top Covering respectively has a hole, inserts respectively feminine gender and positive control serum.
By specific embodiment the present invention is carried out more detailed description below the embodiment:
The quick enzyme immunity detection method of embodiment 1 liver rot
Select the rough antigen of liver fluke solubility for use, substep makes refining antigen by sad method and DEAE52 ion chromatography column chromatography purification, adopts ultraviolet method to measure protein content; Carry out trial test, with other three kinds of antigen contrast experiments, mensuration also selects liver fluke to make with extra care the bag of antigen by concentration; (form PH9.60,0.05M) dilution antigen by sodium bicarbonate and sodium carbonate with carbonate buffer solution, bag is by 96 hole polystyrene microwell plates, 100 microlitres/hole, ambient temperature overnight is got rid of liquid in the hole, the phosphate buffer that contains 1% bovine serum albumin(BSA) with same dosage (is made up of sodium dihydrogen phosphate and sodium hydrogen phosphate, PH7.0,0.05M) the room temperature sealing is 2 hours, vacuum freezedrying, the polybag of packing under the vacuum condition pastes corresponding sign on the bag.Bag also can directly use after being sealed off by plate.
Prepare general liquid reagent, comprise enzyme conjugates liquid B, cleansing solution C, substrate D, developer E, sample diluting liquid F, terminator G, positive reference substance H and negative control product I, enzyme conjugates B liquid phosphate buffer (is made up of sodium dihydrogen phosphate and sodium hydrogen phosphate, PH7.0,0.05M), interpolation stabilizing agent molecular weight Mr is 20000 polyglycol weight 0.5%-5%, add accelerator molecular weight Mr simultaneously and be 6000 polyglycol weight 0.5%-5%, add ten thousand/thimerosal and four kinds of anti-immunoglobulin G while horseradish peroxidase-labeled bonds that parasitic disease is unified.Other composition or collocation method of reagent that detects usefulness is as follows: cleansing solution C: (be made up of sodium dihydrogen phosphate and sodium hydrogen phosphate, PH7.0 0.05M), adds ten thousand/thimerosal to adopt the phosphate buffer of 0.5% tween; Substrate solution D and developer E: substrate solution is a superoxol, and developer is harmless TMB solution; Sample dilution F: (be made up of sodium dihydrogen phosphate and sodium hydrogen phosphate, PH7.0 0.1M), contains tween 0.5% to phosphate buffer; Stop buffer G:1% lauryl sodium sulfate (SDS); Positive control H and negative control I: positive control is with four kinds of positive patients serum inactivation treatment, mixes, and adds the packing of ten thousand/thimerosal, become can be general the mixing positive control; Negative control is after some normal human serums mix, and adds the packing of ten thousand/thimerosal and preserves.Above component has comprised detection needed whole reagent (the distilled water needs are prepared separately), and wherein the equal adjusted of liquid component need not be diluted and can directly use in the working concentration and the plastic dropping bottle of packing into; Plastic tube with cover is put in the negative and positive contrast, can directly use;
Antigen coated microplate and the liquid reagent bottle porous of packing into is successively detected box.It is a kind of kit of four unificationizations that can fasten that porous detects box, be divided into two districts in the box, place four kinds of parasite antigens and wrap in advance by plate in a district, totally four immediate vicinity of fluid reagent bottles, another district is provided with a multiple general liquid reagent bottle of the vertical parallel placement of perforate base, there is opening the kit top, and a hole is respectively arranged on the inside cover, inserts feminine gender and positive control serum respectively.Detecting box need preserve down at 4 ℃, must not be freezing, and the term of validity can reach 6 months with this understanding.
Testing process uses porous to detect box, detailed process comprises: the dilution of (1) sample: with sample dilution F serum to be checked was diluted in 1: 50 by volume, the yin and yang attribute control serum is dilution synchronously in proportion, (2) application of sample reaction then: every Kong Jiayi dilute serum 100ul, establish feminine gender simultaneously, each hole of the positive and blank, the blank hole adds 100ul sample diluting liquid F, 37 ℃ left standstill 10 minutes, get rid of, every hole adds one of cleansing solution C, immediately repeatedly with the distilled water washing, get rid of, pat dry, (3) add enzyme reaction again: add two of enzyme conjugates B, 37 ℃ were reacted 10 minutes, get rid of, add one of cleansing solution C, use the distilled water cyclic washing immediately repeatedly, get rid of, pat dry, (4) chromogenic reaction at last: add each one of substrate D and developer E, mixing was placed 2-3 minute under the room temperature, when treating that negative control hole is little and being blueness, add mixing of terminator G, cessation reaction, visual inspection is shallower than that to be bordering on negative control negative, be and obviously be deeper than the blue positive of negative control, judge with instrument: return to zero with blank and read the O.D value, treat that verify O.D value is positive greater than 2.1 times of persons of negative control, when negative control O.D value is lower than 0.07, calculate by 0.07 in 630nm.
In concrete testing process, must be noted that following problem: 1, kit is preserved down at 2-8 ℃, and elder generation's balance is used to room temperature when taking out at every turn.Drop bottle at every turn with after lid must be tightened, can't use with between each bottle cap; When 2, washing distilled water is filled it up with in the hole, parked for 30 seconds at every turn, get rid of liquid in the hole, other water sources such as forbidding tap water.
The quick enzyme immunity detection method of embodiment 2 paragonimiasis
Select the rough antigen of lung fluke solubility for use, substep makes refining antigen by sad method and DEAE52 ion chromatography column chromatography purification, adopts ultraviolet method to measure protein content; Carry out trial test,, measure and select the refining antigen coated concentration of lung fluke with other three kinds of antigen contrast experiments; With carbonate buffer solution (composition such as above-mentioned, PH9.60,0.05M) dilution antigen, bag is by 96 hole polystyrene microwell plates, 100 microlitres/hole, ambient temperature overnight is got rid of liquid in the hole, phosphate buffer (the composition such as above-mentioned that contains 1% bovine serum albumin(BSA) with same dosage, PH7.0,0.05M) the room temperature sealing is 2 hours, vacuum freezedrying, vacuum condition is pack down, pastes corresponding sign on the plate bag.Bag also can directly use after being sealed off by plate.
Other reagent compound method, be assembled into box mode and detection scheme with embodiment 1.
The quick enzyme immunity detection method of embodiment 3 pork measles
Select the rough antigen of pork measles capsule liquid solubility for use, substep makes refining antigen by sad method and DEAE52 ion chromatography column chromatography purification, adopts ultraviolet method to measure protein content; Carry out trial test,, measure and select antigen coated concentration with other three kinds of antigen contrast experiments; With carbonate buffer solution (composition such as above-mentioned, PH9.60,0.05M) dilution antigen, bag is by 96 hole polystyrene microwell plates, 100 microlitres/hole, ambient temperature overnight is got rid of liquid in the hole, phosphate buffer (the composition such as above-mentioned that contains 1% bovine serum albumin(BSA) with same dosage, PH7.0,0.05M) the room temperature sealing is 2 hours, vacuum freezedrying, vacuum condition is pack down, pastes corresponding sign on the plate bag.Bag also can directly use after being sealed off by plate.
Other reagent compound method, be assembled into box mode and detection scheme with embodiment 1.
The quick enzyme immunity detection method of embodiment 4 trichinosises
Select the rough antigen of trichina muscle phase larva solubility for use, substep makes refining antigen by sad method and DEAE52 ion chromatography column chromatography purification, adopts ultraviolet method to measure protein content; Carry out trial test,, measure and select antigen coated concentration with other three kinds of antigen contrast experiments; With carbonate buffer solution (composition such as above-mentioned, PH9.60,0.05M) dilution antigen, bag is by 96 hole polystyrene microwell plates, 100 microlitres/hole, ambient temperature overnight is got rid of liquid in the hole, phosphate buffer (the composition such as above-mentioned that contains 1% bovine serum albumin(BSA) with same dosage, PH7.0,0.05M) the room temperature sealing is 2 hours, vacuum freezedrying, vacuum condition is pack down, pastes corresponding sign on the plate bag.Bag also can directly use after being sealed off by plate.
Other reagent compound method, be assembled into box mode and detection scheme with embodiment 1.
Claims (4)
1, the quick enzyme immunity detection method of a kind of food-borne parasitic disease, be used for pork measles, lung fluke, liver fluke and trichinous enzyme immune detection, it is characterized in that: the enzyme immune detection comprises antigen coated microplate A, enzyme conjugates liquid B, cleansing solution C, substrate D, developer E, sample diluting liquid F, terminator G, positive reference substance H and negative control product I with reagent, testing process uses porous to detect box, and detailed process comprises:
(1) sample dilution: with serum to be checked dilution in 1: 50 by volume, the yin and yang attribute control serum is dilution synchronously in proportion, then with sample dilution F
(2) application of sample reaction: every Kong Jiayi dilute serum 100ul, establish each hole of feminine gender, the positive and blank simultaneously, the blank hole adds 100ul sample diluting liquid F, and 37 ℃ left standstill 10 minutes, get rid of, every hole adds one of cleansing solution C, immediately with the distilled water washing repeatedly, get rid of, pat dry, again
(3) add enzyme reaction: add two of enzyme conjugates B, 37 ℃ were reacted 10 minutes, got rid of, and added one of cleansing solution C, used the distilled water cyclic washing immediately repeatedly, got rid of, and patted dry, last
(4) chromogenic reaction: add each one of substrate D and developer E, mixing, placed under the room temperature 2-3 minute, and when treating that negative control hole is little and being blueness, added mixing of terminator G, cessation reaction, visual inspection is shallower than that to be bordering on negative control negative, is obviously to be deeper than the blue positive of negative control, judges with instrument: return to zero with blank and read the O.D value in 630nm, treat that verify O.D value is positive greater than 2.1 times of persons of negative control, when negative control O.D value is lower than 0.07, calculate by 0.07.
2, detection method according to claim 1, it is characterized in that: antigen coated microplate A adopts Fast Packet by technology, envelope antigen on the general purpose polystyrene microwell plate in advance, coating buffer be the carbonate buffer solution that contains antigen (form by sodium bicarbonate and sodium carbonate, PH9.60,0.05M), static 12 hours of room temperature, with 1% bovine serum albumin(BSA) sealing 2 hours, vacuum freezedrying, vacuum bagging seals.
3, detection method according to claim 1, it is characterized in that: enzyme conjugates B liquid phosphate buffer (is made up of sodium dihydrogen phosphate and sodium hydrogen phosphate, PH7.O, 0.05M), interpolation stabilizing agent molecular weight Mr is 20000 polyglycol weight 0.5%-5%, add accelerator molecular weight Mr simultaneously and be 6000 polyglycol weight 0.5%-5%, add ten thousand/thimerosal and four kinds of anti-immunoglobulin G while horseradish peroxidase-labeled bonds that parasitic disease is unified.
4, the employed porous of the detection method of claim 1 detects box, it is characterized in that: it is a kind of kit of four unificationizations that can fasten that porous detects box, be divided into two districts in the box, place multiple parasite antigen and wrap in advance by plate in a district, be total to polylith immediate vicinity of fluid reagent bottle, another district is provided with holder of a perforate paper or the multiple general liquid reagent bottle of the vertical parallel placement of plastic base, and there is opening the kit top, a hole is respectively arranged on the inside cover, insert feminine gender and positive control serum respectively.
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| CN 03113564 CN1424585A (en) | 2003-01-15 | 2003-01-15 | Quick enzyme immuno-determining method and box for food borne parasites |
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| CN 03113564 CN1424585A (en) | 2003-01-15 | 2003-01-15 | Quick enzyme immuno-determining method and box for food borne parasites |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102279259A (en) * | 2010-06-10 | 2011-12-14 | 中国疾病预防控制中心寄生虫病预防控制所 | Diagnostic reagent kit for paragonimiasis detection and preparation method thereof |
| CN102914654A (en) * | 2011-08-04 | 2013-02-06 | 董方 | Confining liquid and confining method of biochip for detecting various proteins |
| CN101387641B (en) * | 2007-09-11 | 2013-11-27 | 深圳市康百得生物科技有限公司 | Liver fluke rapid immune detection kit for fresh water fishes and detection method |
-
2003
- 2003-01-15 CN CN 03113564 patent/CN1424585A/en active Pending
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101387641B (en) * | 2007-09-11 | 2013-11-27 | 深圳市康百得生物科技有限公司 | Liver fluke rapid immune detection kit for fresh water fishes and detection method |
| CN102279259A (en) * | 2010-06-10 | 2011-12-14 | 中国疾病预防控制中心寄生虫病预防控制所 | Diagnostic reagent kit for paragonimiasis detection and preparation method thereof |
| CN102914654A (en) * | 2011-08-04 | 2013-02-06 | 董方 | Confining liquid and confining method of biochip for detecting various proteins |
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