CN117917570A - Diagnostic kit for determining immune interference substances and application method thereof - Google Patents
Diagnostic kit for determining immune interference substances and application method thereof Download PDFInfo
- Publication number
- CN117917570A CN117917570A CN202211289144.4A CN202211289144A CN117917570A CN 117917570 A CN117917570 A CN 117917570A CN 202211289144 A CN202211289144 A CN 202211289144A CN 117917570 A CN117917570 A CN 117917570A
- Authority
- CN
- China
- Prior art keywords
- pad
- diagnostic kit
- kit
- colloidal gold
- nitrocellulose membrane
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 239000000126 substance Substances 0.000 title claims abstract description 30
- 238000000034 method Methods 0.000 title claims abstract description 29
- 238000009007 Diagnostic Kit Methods 0.000 title claims abstract description 23
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 claims abstract description 21
- 239000000020 Nitrocellulose Substances 0.000 claims abstract description 16
- 239000012528 membrane Substances 0.000 claims abstract description 16
- 229920001220 nitrocellulos Polymers 0.000 claims abstract description 16
- 239000000853 adhesive Substances 0.000 claims abstract description 7
- 230000001070 adhesive effect Effects 0.000 claims abstract description 7
- 238000010521 absorption reaction Methods 0.000 claims abstract description 5
- 239000002981 blocking agent Substances 0.000 claims description 21
- 210000002966 serum Anatomy 0.000 claims description 16
- 108090000623 proteins and genes Proteins 0.000 claims description 8
- 239000007853 buffer solution Substances 0.000 claims description 7
- 102000004169 proteins and genes Human genes 0.000 claims description 7
- 241000283707 Capra Species 0.000 claims description 6
- 241001465754 Metazoa Species 0.000 claims description 6
- 241000283973 Oryctolagus cuniculus Species 0.000 claims description 6
- 239000002504 physiological saline solution Substances 0.000 claims description 6
- 239000003755 preservative agent Substances 0.000 claims description 6
- 230000002335 preservative effect Effects 0.000 claims description 6
- 239000000463 material Substances 0.000 claims description 4
- 241000287828 Gallus gallus Species 0.000 claims description 3
- 101710116034 Immunity protein Proteins 0.000 claims description 3
- 241001494479 Pecora Species 0.000 claims description 3
- 239000003774 sulfhydryl reagent Substances 0.000 claims description 3
- 239000003398 denaturant Substances 0.000 claims description 2
- 238000004043 dyeing Methods 0.000 claims 1
- 238000001514 detection method Methods 0.000 abstract description 15
- 239000003153 chemical reaction reagent Substances 0.000 abstract description 8
- 230000002452 interceptive effect Effects 0.000 abstract description 6
- 239000000523 sample Substances 0.000 description 16
- 208000030507 AIDS Diseases 0.000 description 8
- 238000012360 testing method Methods 0.000 description 7
- 238000002965 ELISA Methods 0.000 description 3
- 238000011161 development Methods 0.000 description 3
- 238000003317 immunochromatography Methods 0.000 description 3
- 230000009871 nonspecific binding Effects 0.000 description 3
- 102100032282 26S proteasome non-ATPase regulatory subunit 14 Human genes 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 101000590281 Homo sapiens 26S proteasome non-ATPase regulatory subunit 14 Proteins 0.000 description 2
- 239000000427 antigen Substances 0.000 description 2
- 102000036639 antigens Human genes 0.000 description 2
- 108091007433 antigens Proteins 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 238000013399 early diagnosis Methods 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 238000009010 Bradford assay Methods 0.000 description 1
- 238000012286 ELISA Assay Methods 0.000 description 1
- 206010018910 Haemolysis Diseases 0.000 description 1
- 206010034719 Personality change Diseases 0.000 description 1
- 208000027418 Wounds and injury Diseases 0.000 description 1
- 239000002250 absorbent Substances 0.000 description 1
- 230000002745 absorbent Effects 0.000 description 1
- 230000036436 anti-hiv Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 230000000903 blocking effect Effects 0.000 description 1
- 238000007813 chromatographic assay Methods 0.000 description 1
- 238000003776 cleavage reaction Methods 0.000 description 1
- 238000004891 communication Methods 0.000 description 1
- 230000002860 competitive effect Effects 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- 238000007599 discharging Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000010931 gold Substances 0.000 description 1
- 229910052737 gold Inorganic materials 0.000 description 1
- 230000008588 hemolysis Effects 0.000 description 1
- 208000014674 injury Diseases 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 239000006166 lysate Substances 0.000 description 1
- 230000003340 mental effect Effects 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 239000013610 patient sample Substances 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 238000004393 prognosis Methods 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000007017 scission Effects 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 239000012224 working solution Substances 0.000 description 1
Landscapes
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention discloses a diagnostic kit for determining immune interference substances and a use method thereof, wherein the diagnostic kit comprises a reagent kit and a use method thereof, the reagent kit comprises an adhesive pad, a nitrocellulose membrane is arranged on the adhesive pad, an absorption pad is arranged on the right side above the nitrocellulose membrane, a colloidal gold pad is arranged on the left side above the nitrocellulose membrane, and a sample pad is arranged above the colloidal gold pad. The diagnostic kit for determining the immune interference substance and the use method thereof can greatly reduce the false positive rate of HIV, improve the specificity and the accuracy of the kit, and are particularly suitable for HIV colloidal gold rapid detection reagents. The invention can rapidly detect whether the suspected sample contains the amphotropic interfering substance in a short time, and the sample use amount is very small, thereby greatly improving the efficiency and facilitating the observation of the detected sample. The invention can greatly reduce the false positive rate of HIV, improve the specificity and accuracy of the kit, and is especially suitable for HIV colloidal gold rapid detection reagents.
Description
Technical Field
The invention relates to the technical field of AIDS, in particular to a diagnostic kit for determining immune interference substances and a use method thereof.
Background
In recent years, the incidence rate of AIDS is extremely high, and HIV viruses can be hidden in organisms, so that patients can normally live and work between 8 and 9 years without any subjective symptoms, and the research shows that the AIDS is originated in Africa and then brought into the United states by immigration. In order to effectively prevent the epidemic of AIDS and reduce the occurrence of AIDS, the prevention of HIV transmission must be carried out early, and early diagnosis is one of the effective modes for controlling the occurrence rate of AIDS. Early diagnosis and treatment of AIDS are necessary links for improving the prognosis ending of patients, and can also control the epidemic spread of the AIDS. The primary screening test of anti-HIV antibody mainly comprises ELISA and colloidal gold test strip, and the detection sensitivity is greatly improved along with the continuous development of HIV raw materials and detection technology, but about 1-5 per mill or even higher false positives exist, so that great psychological burden and mental injury are caused to patients. The cause of false positives in aids agents is mainly several: 1. the serum contains nonspecific antibodies which bind to other parts except HIV epitope; 2. collecting samples to generate character change and hemolysis; 3. other components that bind to non-specific antibodies in serum, etc., are present in the reagent buffer system.
At present, the traditional HIV diagnostic kit has the defects of higher false positive rate during use, lower accuracy, incapability of discharging interference during use, larger dosage and difficulty in meeting the use requirement.
Based on the above, the present invention designs a diagnostic kit for measuring immune interference substances and a method for using the same, so as to solve the above problems.
Disclosure of Invention
The invention aims to provide a diagnostic kit for determining immune interference substances and a use method thereof, which are used for solving the problems that at present, the traditional HIV diagnostic kit has higher false positive rate during use, lower accuracy, can not discharge interference during use, has larger dosage and is difficult to meet the use requirement.
In order to achieve the above purpose, the present invention provides the following technical solutions:
The kit comprises a sticky pad, a nitrocellulose membrane is arranged on the sticky pad, an absorption pad is arranged on the right side above the nitrocellulose membrane, a colloidal gold pad is arranged on the left side above the nitrocellulose membrane, and a sample pad is arranged above the colloidal gold pad.
As a further scheme of the invention, a stop control line is arranged on the nitrocellulose membrane, and a T line is arranged on the left side of the stop control line.
As a further aspect of the present invention, the use method includes:
Step 1, patient serum is dripped on the sample pad, reaches the colloidal gold pad through a siphon principle, and is combined with a blocking agent and dyed;
And 2, capturing the dyed serum arrival line at the T line and developing the color at the T line through a siphon principle.
As a further aspect of the invention, the blocking agent includes biological blocking agents and chemical blocking agents.
As a further aspect of the present invention, the bio-blocker includes a first buffer solution, physiological saline, a first preservative, animal immune proteins, and rabbit serum.
As a further aspect of the present invention, the chemical blocking agent includes a second buffer solution, physiological saline, a second preservative, PEG, a thiol reagent, and a protein denaturing agent, and the molecular weight of the PEG is 1500-20000.
As a further aspect of the invention, the animal immunity protein comprises mouse IgG, goat IgG, sheep IgG, rabbit IgG, horse IgG, chicken IgY.
Compared with the prior art, the invention has the beneficial effects that:
The diagnostic kit for determining the immune interference substance and the use method thereof can greatly reduce the false positive rate of HIV, improve the specificity and the accuracy of the kit, and are particularly suitable for HIV colloidal gold rapid detection reagents.
The invention can rapidly detect whether the suspected sample contains the amphotropic interfering substance in a short time, and the sample use amount is very small, thereby greatly improving the efficiency and facilitating the observation of the detected sample.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings that are needed for the description of the embodiments will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and that other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.
Fig. 1 is a schematic diagram of a diagnostic kit for measuring an immunointerference substance and a method for using the same according to the present invention.
In the drawings, the list of components represented by the various numbers is as follows:
1. an adhesive pad; 2. a nitrocellulose membrane; 3. a colloidal gold pad; 4. a sample pad; 5. an absorbent pad; 6. a stop control line; 7. t line.
Detailed Description
The following description of the embodiments of the present invention will be made clearly and completely with reference to the accompanying drawings, in which it is apparent that the embodiments described are only some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
In the description of the present invention, it should be noted that, directions or positional relationships indicated by terms "upper", "top", "bottom", "one side", "another side", "front", "rear", "middle", "inside", "top", "bottom", etc., are directions or positional relationships based on the drawings, are merely for convenience of describing the present invention and simplifying the description, and do not indicate or imply that the apparatus or elements referred to must have a specific direction, be configured and operated in a specific direction, and thus should not be construed as limiting the present invention; the terms "first," "second," and "third" are used for descriptive purposes only and are not to be construed as indicating or implying relative importance; furthermore, unless explicitly specified and limited otherwise, the terms "mounted," "connected," and "connected" are to be construed broadly, and may be either fixedly connected, detachably connected, or integrally connected, for example; can be mechanically or electrically connected; can be directly connected or indirectly connected through an intermediate medium, and can be communication between two elements. The specific meaning of the above terms in the present invention will be understood in specific cases by those of ordinary skill in the art.
As shown in FIG. 1, the present invention provides a diagnostic kit for measuring immunointerference material and a method for using the same, wherein,
Fig. 1 is a schematic structural diagram of a diagnostic kit for measuring immune interference substances and a method for using the same according to the present invention, and as can be seen from fig. 1, a diagnostic kit for measuring immune interference substances and a method for using the same are provided, which comprises a kit and a method for using the same, wherein the kit comprises an adhesive pad1, a nitrocellulose membrane 2 is arranged on the adhesive pad1, an absorption pad 5 is arranged on the right side above the nitrocellulose membrane 2, a colloidal gold pad 3 is arranged on the left side above the nitrocellulose membrane 2, and a sample pad 4 is arranged above the colloidal gold pad 3.
The nitrocellulose membrane 2 is provided with a stop control line 6, and the left side of the stop control line 6 is provided with a T line 7.
The using method comprises the following steps:
Step 1, patient serum is dripped on the sample pad 4, reaches the colloidal gold pad 3 through a siphon principle, and is combined with a blocking agent and dyed;
and 2, capturing the dyed serum arrival line at the T line 7 and developing the color at the T line 7 by a siphon principle.
Wherein the blocking agent comprises a biological blocking agent and a chemical blocking agent. The biological blocker comprises a first buffer solution, physiological saline, a first preservative, animal immune proteins and rabbit serum. The chemical blocking agent comprises a second buffer solution, physiological saline, a second preservative, PEG, a sulfhydryl reagent and a protein denaturant, wherein the molecular weight of the PEG is 1500-20000. The animal immunity protein comprises mouse IgG, goat IgG, sheep IgG, rabbit IgG, horse IgG and chicken IgY.
In addition, when in use, the blocking agent mixture has an enhanced effect on color development; the function of the stop control line 6 is to combine with the colloidal gold pad 7 to develop color, if the color development exists, the test experiment is smoothly carried out; the absorption pad 5 is used for absorbing redundant liquid; the test paper strip is used for detecting the heterotrophic interfering substance, if the color is developed, the serum of the patient is indicated to have the interfering substance, and the blocking agent is required to be added into the test paper box when the patient is detected later, so that the interference of the interfering substance is discharged, and the detection is accurate.
In the practical application of the present invention,
① When the colloidal gold immunochromatography detection is used for double antigen sandwich method, the colloidal gold immunochromatography detection is added into a sample pad in proper concentration, and substances which cause nonspecific binding in false positive serum can be neutralized when serum is added for detection, so that the generation of false positive results is avoided.
② When the ELISA method is used for detecting double antigen sandwich method, the ELISA method is added into a sample diluent in proper concentration, and when serum is added for detection, substances which cause nonspecific binding in false positive serum can be neutralized, so that false positive results are avoided.
In a first embodiment of the present invention,
Use of an HIV specific blocker in body gold immunochromatography:
The mutant A, B, C, D and E is designed, then the gene is synthesized, subcloned, transformed, expressed and crude purified, and the cleavage supernatant is obtained. Protein concentration was measured using the Bradford method with BSA as standard, the lysate was diluted to 0.5mg/ml with 0.5% SDS, and the sample pad was soaked for 5-10min at 37℃and dried for 3h.
The invention can be used for detecting modes of monoclonal antibody or polyclonal antibody+polyclonal antibody. The customer may choose according to the capture antibodies in the detection reagent and the actual situation of the detection antibody, for example, a sandwich or competitive ELISA or chromatographic assay using goat antibodies should use goat Igs. Any anti-goat antibody in the patient sample that can potentially interfere with the test signal can be conjugated to goat Ig, avoiding non-specific binding that would interfere with the test results. When the blocking amount of the passive blocking agent is selected, it is recommended that the amount of the antibody used in the kit is 15 times or more (mainly referred to as interference caused by RF), and the amount of the enzyme conjugate antibody is usually used as a reference standard, for example, the concentration of the detection antibody is 6ug/ml, the working concentration is 90ug/ml to 100ug/m, and in some samples with particularly strong ampholytic interference, the blocking agent is required to be increased to 200ug/ml.
Or is suitable for a detection mode of a double monoclonal antibody sandwich or a competition or indirect method. The sample is diluted or the enzyme working solution is added. When the passive blocking agent of the present type is used, the recommended amount is more than 10 times that of the antibody of the kit, and in the above case, the recommended working concentration is 50ug/ml to 60ug/ml, and in some samples with particularly strong ampholytic interference, the blocking agent is required to be used in an amount of 100ug/ml to 120ug/ml.
The diagnostic kit for determining the immune interference substance and the use method thereof can greatly reduce the false positive rate of HIV, improve the specificity and the accuracy of the kit, and are particularly suitable for HIV colloidal gold rapid detection reagents.
Furthermore, it was found in the study that some samples of the amphotropic interference do not contain only one type of interference, but two or more types, so that in this case not all interference types can be completely eliminated with a single blocker, requiring the developer to match different combinations of blockers depending on the interference sample encountered.
The invention can rapidly detect whether the suspected sample contains the amphotropic interfering substance in a short time, and the sample use amount is very small, thereby greatly improving the efficiency and facilitating the observation of the detected sample.
The preferred embodiments of the invention disclosed above are intended only to assist in the explanation of the invention. The preferred embodiments are not exhaustive or to limit the invention to the precise form disclosed. Obviously, many modifications and variations are possible in light of the above teaching. The embodiments were chosen and described in order to best explain the principles of the invention and the practical application, to thereby enable others skilled in the art to best understand and utilize the invention. The invention is limited only by the claims and the full scope and equivalents thereof.
Claims (7)
1. The diagnostic kit for determining immune interference substances comprises a kit and a using method, and is characterized in that the kit comprises an adhesive pad (1), a nitrocellulose membrane (2) is arranged on the adhesive pad (1), an absorption pad (5) is arranged on the right side above the nitrocellulose membrane (2), a colloidal gold pad (3) is arranged on the left side above the nitrocellulose membrane (2), and a sample pad (4) is arranged above the colloidal gold pad (3).
2. The diagnostic kit for measuring immune interference substances and the use method thereof according to claim 1, wherein a stop control line (6) is arranged on the nitrocellulose membrane (2), and a T line (7) is arranged on the left side of the stop control line (6).
3. The diagnostic kit for measuring immunointerference material according to claim 1 and 2, and the use method thereof, characterized in that the use method comprises:
Step 1, patient serum is dripped on the sample pad (4), and reaches the colloidal gold pad (3) through a siphon principle, and is combined with a blocking agent for dyeing;
And 2, capturing the dyed serum arrival line at the T line (7) by a siphon principle, and developing.
4. A diagnostic kit for determining immunointerference material and its method of use according to claim 3, characterized in that said blocking agent comprises a biological blocking agent and a chemical blocking agent.
5. The diagnostic kit for measuring immune interference substance and the use method thereof according to claim 4, wherein the bio-blocker comprises a first buffer solution, physiological saline, a first preservative, animal immune protein, and rabbit serum.
6. The diagnostic kit for measuring immune interference substances and the use method thereof according to claim 4, wherein the chemical blocking agent comprises a second buffer solution, physiological saline, a second preservative, PEG, a sulfhydryl reagent and a protein denaturant, and the molecular weight of the PEG is 1500-20000.
7. The diagnostic kit for determining immunointerference material of claim 5, wherein said animal immunity protein comprises mouse IgG, goat IgG, sheep IgG, rabbit IgG, horse IgG, chicken IgY.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211289144.4A CN117917570A (en) | 2022-10-20 | 2022-10-20 | Diagnostic kit for determining immune interference substances and application method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211289144.4A CN117917570A (en) | 2022-10-20 | 2022-10-20 | Diagnostic kit for determining immune interference substances and application method thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN117917570A true CN117917570A (en) | 2024-04-23 |
Family
ID=90729769
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202211289144.4A Pending CN117917570A (en) | 2022-10-20 | 2022-10-20 | Diagnostic kit for determining immune interference substances and application method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN117917570A (en) |
-
2022
- 2022-10-20 CN CN202211289144.4A patent/CN117917570A/en active Pending
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US11959912B2 (en) | Fluorescence immunochromatographic detection card and a preparation method therefor and use thereof | |
| Grauballe et al. | Optimized enzyme‐linked immunosorbent assay for detection of human and bovine rotavirus in stools: Comparison with electron‐microscopy, immunoelectro‐osmophoresis, and fluorescent antibody techniques | |
| Yorde et al. | Competitive enzyme-liked immunoassay with use of soluble enzyme/antibody immune complexes for labeling. I. Measurement of human choriogonadotropin. | |
| US20120015350A1 (en) | Lateral flow strip and uses thereof | |
| Hogrefe et al. | Detection of herpes simplex virus type 2-specific immunoglobulin G antibodies in African sera by using recombinant gG2, Western blotting, and gG2 inhibition | |
| US7785818B2 (en) | Inflammatory bowel disease and irritable bowel syndrome IBD-first chek diagnostic panel | |
| CN111233985A (en) | Preparation method of novel coronavirus IgA antibody rapid detection test strip | |
| CN112964884B (en) | Diagnostic marker and application thereof in COVID-19 diagnosis and coronavirus past infection detection | |
| Gambino et al. | Comparison of a rapid immunochromatographic test with a chemiluminescence immunoassay for detection of anti-SARS-CoV-2 IgM and IgG | |
| JPWO2015080286A1 (en) | Detection method using immunochromatography | |
| EP0291086A2 (en) | Method for the determination of an antibody in human body fluids | |
| CA3181751A1 (en) | Detection of antibodies to sars-cov-2 | |
| WO2008012650A2 (en) | An immunochromatography device for the diagnosis of diseases in a sample | |
| CN115184620B (en) | Quantum dot fluorescence detection test strip and kit for PLA2R antibody and application of quantum dot fluorescence detection test strip and kit | |
| Patzl et al. | Determination of autoantibodies to thyroglobulin, thyroxine and triiodothyronine in canine serum | |
| CN113490684A (en) | Monoclonal antibodies specific for the PB2 antigen of the human influenza virus (FLU), nucleotide sequences, methods and kits for diagnosing infections produced by FLU | |
| CN117917570A (en) | Diagnostic kit for determining immune interference substances and application method thereof | |
| Chin et al. | Heterophile antibody interference with thyroid assay | |
| CN108267594A (en) | A kind of ST2 detection kits, method of preparation and use based on bimolecular fluorescence complementary technology | |
| Decoster et al. | Bicentric evaluation of Access Toxo immunoglobulin M (IgM) and IgG assays and IMx toxo IgM and IgG assays and comparison with Platelia Toxo IgM and IgG assays | |
| JP5857385B2 (en) | Bladder cancer diagnostic composition containing APE1 / REF-1 and bladder cancer diagnostic kit using the same | |
| JP6357425B2 (en) | Interfering peptide and method for detecting microorganisms | |
| CN203191382U (en) | Immunoblotting kit for detecting multiple antinuclear antibodies | |
| CN103323602A (en) | Double antibody sandwich ELISA detection method for TRPC6 protein and kit thereof | |
| Vafai et al. | Rapid, sensitive, and specific lateral-flow immunochromatographic point-of-care device for detection of varicella-zoster virus immunoglobulin G antibodies in fingerstick blood |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |