CN203144418U - Immuno-PCR gene detection kit for escherichia coli O157:H7 - Google Patents
Immuno-PCR gene detection kit for escherichia coli O157:H7 Download PDFInfo
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- CN203144418U CN203144418U CN 201220684769 CN201220684769U CN203144418U CN 203144418 U CN203144418 U CN 203144418U CN 201220684769 CN201220684769 CN 201220684769 CN 201220684769 U CN201220684769 U CN 201220684769U CN 203144418 U CN203144418 U CN 203144418U
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Abstract
The utility model relates to an immuno-PCR gene detection kit for escherichia coli O157:H7, belongs to the field of detection of food-borne pathogenic bacteria, and is suitable for rapid detection of escherichia coli O157:H7 in various food products. The detection kit comprises a kit cover (1), a kit body (2), a plastic base plate (3), a first reagent unit, a second reagent unit, and a self-sealing bag (12). The first reagent unit is a molecular biological reagent; 10*PCR buffer (4), dNTP (5), TaqDNA polymerase (6), ddH2O double distilled water (7), a forward primer (8), and a reverse primer (9) are respectively filled in EP tubes; reagent bottles of the second reagent unit are respectively filled with a positive control solution (10), and a negative control solution (11); the self-sealing bag is filled with a detection immune-PCR tube coated with escherichia coli O157:H7 monoclonal antibodies. The kit cover is connected with the kit body; the first reagent unit and the second reagent unit are orderly inlayed on the plastic base plate; the self-sealing bag is disposed on the plastic base plate; and the plastic base plate is disposed in the kit box. The immunocapture PCR detection kit for escherichia coli O157:H7 of the utility model combines principles of immunology and molecular technology, and can rapidly, accurately, sensitively, and directly detect pathogenic bacteria in food.
Description
Technical field
The utility model relates to a kind of Escherichia coli O 157: H7 (Escherichia coli O157:H7) immuno-PCR gene detecting kit, belong to the food-borne pathogens detection range, specially at the detection of food-borne pathogens Escherichia coli O 157: H7 (Escherichia coli O157:H7).This test kit is applicable to the rapid detection of Escherichia coli O 157 in the various food: H7.
Background technology
Food-borne pathogens is the most serious global food-safety problem, the statistical result showed in 2003～2004 years, and China's food poisoning paathogenic factor is followed successively by microorganism property, chemical, poisonous of animal or plant nature cause of disease.The microorganism venereal disease is former to be to cause the principal element of poisoning by food, and the poisoning number is maximum.The food-borne pathogens rapid detection is the focus that research is paid close attention to always.
The intestinal tract infections disease that EHEC (EHEC) O157:H7 (Escherichia coli O157:H7) causes has become a serious global public health problem, and this disease can cause diarrhoea, hemorrhagic enteritis, secondary hemolytic uremic syndrome (HUS), thrombotic thrombocytopenic purpura (TTP) etc.The state of an illness of HUS and TTP is dangerous, and wherein 3%～5% HUS patient death has 12% patient HUS that serious sequela is arranged approximately, endangers very serious.Since nineteen eighty-two, the U.S. found this disease first, many countries have taken place to break out with popular in succession in the world, especially Japan has taken place by Escherichia coli O 157 between 5～August in 1996: biggest one outbreak of epidemic on the human history that H7 causes, accumulation patient nearly 10000 people, and several people's death are arranged, caused the common concern in the world.China just found in 1987 microbially thus to be dispersed in infection, had more than ten province to detect this pathogenic bacterium in recent years successively in food, poultry, domestic animal, insect and diarrhea cases, existed that epidemic situation is broken out, popular potential threat.
The traditional method that detects Escherichia coli O 157: H7 at present mainly contains biochemical reaction and serotype etc.Biochemical method is loaded down with trivial details, time-consuming, and is subjected to all multifactor impacts, easy omission natural mutant, but and the bacterium that can not detect non-cultivation conditions alive.The serological method development is very fast, and sensitivity is also higher, but can only do the retrospective diagnosis or indirect diagnosis basis is provided, and can not carry out Rapid identification.Therefore, setting up fast and accurately, diagnostic method has become clinical active demand.(immuno polymerase chain reaction IM-PCR) is a kind of highly sensitive technology that detects micro-antigen to immuno-PCR.This technology combines the hypersensitivity of the high specific of antigen antibody reaction and polymerase chain reaction, and its essence is a kind ofly to replace enzyme reaction to amplify a kind of modified form ELISA of antigen-antibody combination rate with pcr amplification section of DNA reporter molecules.Immuno-PCR is the most responsive so far a kind of antigen detection method, can detect single antigen molecule in theory, but its susceptibility is subjected to the influence of many factors in the practice, as the concentration of the selection that connects molecule, indicating system, DNA reporter molecules, PCR cycle index etc.At present, generally the ELI SA method than existing is high by 10 for the susceptibility of the immuno-PCR of reporting both at home and abroad
2～10
8Doubly.
The utility model is a kind of Escherichia coli O 157: H7 immunocapture PCR detection kit, come rapid detection Escherichia coli O 157: H7 with this.
The utility model content
At the deficiencies in the prior art, the purpose of this utility model is to set up a kind of quick, can fast detect the detection kit of food-borne pathogens Escherichia coli O 157: H7 accurately.With pathogenic bacterium immunity enrichment method and the effective combination of gene level detection method.Earlier with specific antibody with the pathogenic bacteria specific immunity enrichment in the sample, pcr amplification has not only largely improved detection sensitivity then, and immunity improved the specificity that detects in conjunction with pcr amplification then, has improved the efficient that pathogenic bacteria detects greatly.
The technical scheme that the utility model is taked is such Escherichia coli O 157: H7 immunocapture PCR detection kit comprises lid (1), box body (2), plastic bottom board (3), first reagent unit, second reagent unit, valve bag (12); First reagent unit is molecular biology reagent, and that adorn respectively in the EP pipe is 10 * PCR Buffer (4), dNTP (5), TaqDNA polysaccharase (6), ddH
2O distilled water (7), forward primer (8), reverse primer (9); Adorn respectively in the second reagent unit reagent bottle and have plenty of positive control solution (10), negative control solution (11); Adorn in the valve bag to have had plenty of and wrap by Escherichia coli O 157: the detection immuno-PCR pipe of H7 monoclonal antibody; Lid is connected with box body, and first reagent unit and second reagent unit are embedded on the plastic bottom board successively, and valve bag is positioned on the plastic bottom board, and plastic bottom board is positioned in the box body.。
Further, in the described molecular biology reagent, 10 * PCR Buffer loading amount is 300 μ L; The dNTP loading amount is 150 μ L; TaqDNA polysaccharase loading amount is 60 μ L; The forward primer loading amount is 150 μ L; Reverse primer is that loading amount is 150 μ L.
Further, described positive control solution is the Escherichia coli O 157 of deactivation: H7 bacterium liquid, negative control solution increases bacterial context soup for the improvement E.C Vulkamycin. PA-93 of sterilization.
The beneficial effects of the utility model: Escherichia coli O 157: H7 immunocapture PCR detection kit is a kind of food-borne pathogens detection kit, belongs to the food-borne pathogens detection range, is applicable to the rapid detection of Escherichia coli O 157 in the various food: H7.This test kit collection pathogenic bacteria concentrates, specific antibody identification, and pcr amplification is one, has advantages such as detected result accuracy height, highly sensitive, detection be quick, easy to use.Be applicable to fields such as bacterial strain Rapid identification, the detection of foodborne illness source, be particularly useful for the rapid detection of micro-cause of disease in the sample.Be applicable to quality supervision department, import and export sanitary authority etc. carries out Escherichia coli O 157 to the food sample: the evaluation of H7 bacterium.
Escherichia coli O 157: H7 immunocapture PCR detection kit detection accuracy height, Escherichia coli O 157 in the sample: H7 is through the identification of specific antibody, carry out pcr amplification afterwards, pathogenic bacterium are through the double check of immunology, two kinds of detection techniques of molecular biology, guarantee the accuracy that detects, avoided the false positive in the PCR detection.Has highly sensitive, monoclonal antibody on the PCR tube wall catch and enrichment the Escherichia coli O 157 in the sample: H7, improved the template amount indirectly, improved detection sensitivity greatly, after tested, this test kit can reach 10 to the detection sensitivity of Escherichia coli O 157: H7
2Therefore cfu/mL, it is not serious to be particularly suitable for sample contamination, the sample that germ is less.Can once identify concrete kind, after the cause of disease that monoclonal antibody catches, will belong to Escherichia coli O 157 not of the same race together accurately by PCR: the H7 one-time detection is to planting.Avoid assorted bacterium to disturb, the monoclonal antibody on the tube wall is only identified the Escherichia coli O 157 in the sample: H7, is removed in the washing behind other non-object bacteria antibody capture, therefore greatly reduces the interference of assorted bacterium.Need not to extract DNA, the object bacteria loss is little, and all experiment is finished in same PCR pipe, has reduced the cause of disease loss in the conventional PCR detection amplifying nucleic acid leaching process.Detect fast, all detection can be finished at 4 hours.Therefore be a kind of quick, easy, accurate, highly sensitive detection Escherichia coli O 157: the test kit of H7.
The checking research of test kit detection specificity: this test kit is to Escherichia coli O 157: the H7 specificity detects.Specific antibody identification, the PCR condition of Auele Specific Primer rfbF and optimization increases to object bacteria, adopt the non-object bacteria of 10 classes to detect simultaneously, the result shows to have only Escherichia coli O 157: H7 that the PCR product of target stripe 499bp is arranged, other the non-object bacteria of 10 classes all driftlessness band occurs, and has shown the specificity of test kit.
The checking research of test kit detection sensitivity: this test kit is to Escherichia coli O 157: H7 sensitivity detects.It is 1.04 * 10 that the direct PCR of the pure bacterium liquid of Escherichia coli O 157: H7 detects minimum detectability
5Cfu/mL, and the minimum detectability of immunocapture PCR is 1.04 * 10
2Cfu/mL, for pure bacterium liquid, immunocapture PCR sensitivity is 1000 times of direct PCR sensitivity, is 1000 times of ELISA detection sensitivity.This test kit detects medium sensitivity at the food sample and reaches 1.04 * 10
3Cfu/mL.
Rely on the Escherichia coli O 157 of PCR: H7 immunocapture detection kit, related to a kind of molecular engineering that detects the high risk food-borne pathogens, overcome the time-consuming consumption power of existing detection food-borne pathogens detection technique, the low shortcoming that waits of sensitivity.Test kit of the present utility model can be quick, accurate, sensitive from food, directly detect pathogenic bacteria.Simplified trace routine greatly, be applicable to quality supervision department, import and export sanitary authority etc. carries out Escherichia coli O 157 to the food sample: the evaluation of H7 bacterium.
Description of drawings
Fig. 1 is structural representation of the present utility model
Fig. 2 is Escherichia coli O 157: H7 immunocapture detection kit is used flow process and schematic diagram;
Embodiment
Below in conjunction with embodiment and accompanying drawing the utility model is described in further detail and completely, institute gives an actual example and only is used for explaining the utility model, is not for limiting scope of the present utility model.
1 one kinds of Escherichia coli O 157s of embodiment: H7 immunocapture detection kit
Comprise lid (1), box body (2), plastic bottom board (3), first reagent unit, second reagent unit, valve bag (12).First reagent unit is molecular biology reagent, and that adorn respectively in the EP pipe is 10 * PCR Buffer (4), dNTP (5), TaqDNA polysaccharase (6), ddH
2O distilled water (7), forward primer (8), reverse primer (9); Adorn respectively in the second reagent unit reagent bottle and have plenty of positive control solution (10), negative control solution (11); Adorn in the valve bag to have had plenty of and wrap by Escherichia coli O 157: the detection immuno-PCR pipe of H7 monoclonal antibody.Lid is connected with box body, and first reagent unit and second reagent unit are embedded on the plastic bottom board successively, and valve bag is positioned on the plastic bottom board, and plastic bottom board is positioned in the box body.
Described Escherichia coli O 157: the concrete method for coating of the detection immuno-PCR pipe of H7 monoclonal antibody is:
1., dilute Escherichia coli O 157 with coating buffer: H7 monoclonal antibody to 1: 300 times, antibody concentration is 10 μ g/mL, and the every pipe 50 μ L of monoclonal antibody that dilution is good add in the PCR pipe 4 ℃ of bag quilts that spend the night;
2., wrapping the prescription that is cushioned liquid (Carbonate Coating buffer (1 *)) is: anhydrous sodium carbonate Na
2CO
31.59g, sodium bicarbonate NaHCO
32.93g, sodiumazide NaN
30.2g, be dissolved in the 1000ml distilled water, transfer pH to 9.6,4 ℃ of preservations.
The direct PCR research experiment of the pure bacterium liquid of embodiment 2 gradient dilution Escherichia coli O 157: H7
Escherichia coli O 157: the H7 reference culture with stroke-physiological saline solution 10 times of dilutions of successively decreasing successively, is labeled as 10 after cultivating 18h then successively
-1, 10
-2, 10
-3, 10
-4, 10
-5, 10
-6, 10
-7, with 10
-5, 10
-6, 10
-7The bacterium liquid of gradient carries out plate count.The original bacterial concentration of enumeration is 1.04 * 10
9Cfu/mL dilutes original bacteria liquid successively, and the bacterial concentration gradient is: 1.04 * 10
8～1.04 * 10cfu/mL.Each the 5 μ L of bacterium liquid that get each gradient add in the PCR pipe, add No. 4 reagent 5 μ L in the utility model test kit then, No. 5 reagent 2 μ L, and No. 8 reagent 2 μ L, No. 9 reagent 2 μ L, No. 6 reagent 1 μ L, No. 7 reagent is supplied volume to 50 μ L.The amplification condition of PCR is 95 ℃ of 5min; 35 circulation (95 ℃ of 15s; 55 ℃ of 30s; 72 ℃ of 1min), 72 ℃ of 5min, 4 ℃ of preservations.The PCR product of getting 10 μ L detects target stripe and gel imaging analysis with 1.5% agarose gel electrophoresis.
Among the gel imaging result, the M swimming lane is 1000bp DNA ladder; The negative contrast of N swimming lane; The bacterial concentration of 1-8 swimming lane correspondence is: 1.04 * 10
8, 1.04 * 10
7, 1.04 * 10
6, 1.04 * 10
5, 1.04 * 10
4, 1.04 * 10
3, 1.04 * 10
2, 1.04 * 10cfu/mL.Have for swimming lane 1-4 number the specificity target stripe to occur, clip size is 499bp, and brightness weakens successively.5-8 swimming lane and negative control do not have specific band.The result shows that the sensitivity that the direct PCR of the pure bacterium liquid of Escherichia coli O 157: H7 detects is 1.04 * 10
5Cfu/mL.
This test kit of the pure bacterium liquid of embodiment 3 gradient dilution Escherichia coli O 157: H7 detects the IC-PCR research experiment
Get the Escherichia coli O 157 of gradient dilution: (1.04 * 108～1.04 * 10cfu/mL) each 200 μ L place the detection PCR pipe in this test kit to H7 bacterium liquid, be positioned over 37 ℃ and hatch 2h, remove bacterium liquid with careful suction of pipettor then, it is inferior to give a baby a bath on the third day after its birth with PBST, at the dried raffinate of aseptic filter paper button, in the PCR pipe, add No. 4 reagent 5 μ L in the utility model test kit then, No. 5 reagent 2 μ L, No. 8 reagent 2 μ L, No. 9 reagent 2 μ L, No. 6 reagent 1 μ L, No. 7 reagent is supplied volume to 50 μ L.The amplification condition of PCR is 95 ℃ of 5min; 35 circulation (95 ℃ of 15s; 55 ℃ of 30s; 72 ℃ of 1min), 72 ℃ of 5min, 4 ℃ of preservations.The PCR product of getting 10 μ L detects target stripe and gel imaging analysis with 1.5% agarose gel electrophoresis.
Among the gel imaging result, the M swimming lane is 1000bp DNA ladder; The negative contrast of N swimming lane; The bacterial concentration of 1-8 swimming lane correspondence is: 1.04 * 10
8, 1.04 * 10
7, 1.04 * 10
6, 1.04 * 10
5, 1.04 * 10
4, 1.04 * 10
3, 1.04 * 10
2, 1.04 * 10cfu/mL.Have for swimming lane 1-7 number the specificity target stripe to occur, clip size is 499bp, and brightness weakens successively.8 swimming lanes and negative control do not have specific band.The result shows, Escherichia coli O 157: the sensitivity that H7 immuno-PCR detection kit detects the pure bacterium liquid of Escherichia coli O 157: H7 detectability is 1.04 * 10
2Cfu/mL.
4 test kits of embodiment are to Escherichia coli O 157: H7 and non-Escherichia coli O 157: the experiment of H7 The specificity,
Strains tested has: Escherichia coli O 157: H7ATCC82346, streptococcus aureus ATCC27660, Shu Shi salmonella typhi ATCC22956, Salmonella enteritidis ATCC13076, Listeria monocytogenes ATCC43251, yersinia entero-colitica ATCC23715, shigella flexneri ATCC12022, bacillus ceylonensis A ATCC25931, shigella dysenteriae ATCC51329, beta hemolytic streptococcus ATCC10373, Enterobacter sakazakii ATCC29004.Respectively get in the detection PCR pipe that strains tested 200 μ L add this test kit, be positioned over 37 ℃ and hatch 2h, remove bacterium liquid with careful suction of pipettor then, it is inferior to give a baby a bath on the third day after its birth with PBST, at the dried raffinate of aseptic filter paper button, in the PCR pipe, add No. 4 reagent 5 μ L in the utility model test kit then, No. 5 reagent 2 μ L, No. 8 reagent 2 μ L, No. 9 reagent 2 μ L, No. 6 reagent 1 μ L, No. 7 reagent is supplied volume to 50 μ L.The amplification condition of PCR is 95 ℃ of 5min; 35 circulation (95 ℃ of 15s; 55 ℃ of 30s; 72 ℃ of 1min), 72 ℃ of 5min, 4 ℃ of preservations.The PCR product of getting 10 μ L detects target stripe and gel imaging analysis with 1.5% agarose gel electrophoresis.
Result of implementation has only the swimming lane of large intestine O157 bacterium purpose band (499bp) to occur, and other 10 strain control strains all do not have the purpose band to occur, and illustrate that this test kit has excellent specificity.
According to GB GB/T4789.36-2008 " microbiological test of food hygiene colon bacillus O157:H7/NM check ", getting food sample 25g (mL) joins 225mL improvement E.C Vulkamycin. PA-93 and increases among bacterial context soup m (EC) n, bouncing homogeneous 2～3min on the formula homogenizer, as food homogenate, then to the Escherichia coli O 157 of homogenate artificial contamination different concns gradient: the pure bacterium liquid of H7, Escherichia coli O 157 in artificial contamination's the food homogenate: the H7 bacterial concentration is followed successively by: 1.04 * 10
8～1.04 * 10
2Cfu/mL.Each the 5 μ L of food homogenate that get the artificial contamination of different gradients add in the PCR pipe, add No. 4 reagent 5 μ L in the utility model test kit then, No. 5 reagent 2 μ L, No. 8 reagent 2 μ L, No. 9 reagent 2 μ L, No. 6 reagent 1 μ L, No. 7 reagent is supplied volume to 50 μ L.The amplification condition of PCR is 95 ℃ of 5min; 35 circulation (95 ℃ of 15s; 55 ℃ of 30s; 72 ℃ of 1min), 72 ℃ of 5min, 4 ℃ of preservations.The PCR product of getting 10 μ L detects target stripe and gel imaging analysis with 1.5% agarose gel electrophoresis.
Result of study obtains food samples and simulates the Escherichia coli O 157 that carries disease germs: the sensitivity of the direct PCR of H7 can reach 1.04 * 10
5～1.04 * 10
6Cfu/mL.
Embodiment 6 gradient dilutions are simulated the Escherichia coli O 157 that carries disease germs: this detection kit of H7 IC-PCR research experiment
According to GB GB/T4789.36-2008 " microbiological test of food hygiene colon bacillus O157:H7/NM check ", getting food sample 25g (mL) joins in the 225mL physiological saline, bouncing homogeneous 2～3min on the formula homogenizer, as food homogenate, then to the Escherichia coli O 157 of homogenate artificial contamination different concns gradient: the pure bacterium liquid of H7, Escherichia coli O 157 in artificial contamination's the food homogenate: the H7 bacterial concentration is followed successively by: 1.04 * 10
8～1.04 * 10
2Cfu/mL.Each the 200 μ L of food homogenate that get the artificial contamination of different gradients add in the PCR pipe, be positioned over 37 ℃ and hatch 2h, remove bacterium liquid with careful suction of pipettor then, it is inferior to give a baby a bath on the third day after its birth with PBST, at the dried raffinate of aseptic filter paper button, in the PCR pipe, add No. 4 reagent 5 μ L in the utility model test kit then, No. 5 reagent 2 μ L, No. 8 reagent 2 μ L, No. 9 reagent 2 μ L, No. 6 reagent 1 μ L, No. 7 reagent is supplied volume to 50 μ L.The amplification condition of PCR is 95 ℃ of 5min; 35 circulation (95 ℃ of 15s; 55 ℃ of 30s; 72 ℃ of 1min), 72 ℃ of 5min, 4 ℃ of preservations.The PCR product of getting 10 μ L detects target stripe and gel imaging analysis with 1.5% agarose gel electrophoresis.
Result of study obtains Escherichia coli O 157: the sensitivity that the simulation of H7 immunocapture PCR detection kit research food samples is carried disease germs can reach 1.04 * 10
3～1.04 * 10
4Cfu/mL.
Escherichia coli O 157: H7 immunocapture PCR detection kit is easy and simple to handle, detects fast, and highly sensitive and high specific need not extract DNA.Artificial contamination's food samples need not to increase bacterium and directly detects in experiment, and detectability can reach 10
4Cfu/mL, the sample that has reaches 10
3Cfu/mL, sensitivity is 10～100 times of direct PCR.This test kit can directly detect sample, has the bigger basis of applying.
Above only narrate is preferred embodiment of the present utility model, and be all within spirit of the present utility model and principle not in order to limit the utility model, any modification of doing, is equal to replacement, improvement etc., all should be included within protection scope of the present invention.
Claims (3)
1. the immuno-PCR gene detecting kit of Escherichia coli O 157 a: H7 is characterized in that comprising lid (1), box body (2), plastic bottom board (3), first reagent unit, second reagent unit, valve bag (12); First reagent unit is molecular biology reagent, and that adorn respectively in the EP pipe is 10 * PCR Buffer (4), dNTP (5), TaqDNA polysaccharase (6), ddH
2O distilled water (7), forward primer (8), reverse primer (9); Adorn respectively in the second reagent unit reagent bottle and have plenty of positive control solution (10), negative control solution (11); Adorn in the valve bag to have had plenty of and wrap by Escherichia coli O 157: the detection immuno-PCR pipe of H7 monoclonal antibody; Lid is connected with box body, and first reagent unit and second reagent unit are embedded on the plastic bottom board successively, and valve bag is positioned on the plastic bottom board, and plastic bottom board is positioned in the box body.
2. the immuno-PCR gene detecting kit of Escherichia coli O 157 according to claim 1: H7, it is characterized in that: in the described molecular biology reagent, 10 * PCR Buffer loading amount is 300 μ L; The loading amount of dNTP is 150 μ L; TaqDNA polysaccharase loading amount is 60 μ L; The forward primer loading amount is 150 μ L; The reverse primer loading amount is 150 μ L.
3. the immuno-PCR gene detecting kit of Escherichia coli O 157 according to claim 1: H7, it is characterized in that: described positive control solution is the Escherichia coli O 157 of deactivation: H7 bacterium liquid, negative control solution increases bacterial context soup for the improvement E.C Vulkamycin. PA-93 of sterilization.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103823060A (en) * | 2014-01-22 | 2014-05-28 | 上海理工大学 | Immune PCR (Polymerase Chain Reaction) detection kit of hemorrhagic escherichia coli O157:H7 |
| CN103820549A (en) * | 2014-02-25 | 2014-05-28 | 上海理工大学 | Salmonella choleraesuis immune PCR detection kit |
| CN103866033A (en) * | 2014-04-02 | 2014-06-18 | 上海理工大学 | Staphylococcus aureus immune PCR (polymerase chain reaction) detection kit |
-
2012
- 2012-12-11 CN CN 201220684769 patent/CN203144418U/en not_active Expired - Lifetime
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103823060A (en) * | 2014-01-22 | 2014-05-28 | 上海理工大学 | Immune PCR (Polymerase Chain Reaction) detection kit of hemorrhagic escherichia coli O157:H7 |
| CN103823060B (en) * | 2014-01-22 | 2015-11-25 | 上海理工大学 | Enterohemorrhagic Escherichia coli O 157: H7 immuno-PCR detection kit |
| CN103820549A (en) * | 2014-02-25 | 2014-05-28 | 上海理工大学 | Salmonella choleraesuis immune PCR detection kit |
| CN103866033A (en) * | 2014-04-02 | 2014-06-18 | 上海理工大学 | Staphylococcus aureus immune PCR (polymerase chain reaction) detection kit |
| CN103866033B (en) * | 2014-04-02 | 2015-07-15 | 上海理工大学 | Staphylococcus aureus immune PCR (polymerase chain reaction) detection kit |
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| CX01 | Expiry of patent term |
Granted publication date: 20130821 |
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Addressee: Patent of Shanghai Huiyun Biotechnology Co.,Ltd. The person in charge Document name: Notice of termination upon expiration of patent right |