JPWO2019246289A5

JPWO2019246289A5 -

Info

Publication number: JPWO2019246289A5
Application number: JP2020571469A
Authority: JP
Publication date: 2022-06-23

Claims

How to:
a) The first diluted sample is brought into contact with the first aptamer, and the first aptamer affinity complex causes the first aptamer to interact with the target molecule thereof, so that the target molecule is contained in the first diluted sample. The contact, which is formed when present in.
b) The second diluted sample is brought into contact with the second aptamer, and the second aptamer affinity complex causes the target molecule to be contained in the second diluted sample by the interaction between the second aptamer and the target molecule thereof. The contact, which is formed when present in.
c) The third diluted sample is brought into contact with the third aptamer, and the third aptamer affinity complex causes the target molecule to be contained in the third diluted sample by the interaction between the third aptamer and the target molecule thereof. The contact, which is formed when present in.
d) Incubate the first and second diluted samples separately to allow the formation of an aptamer-affinity complex;
e) Incubate the third diluted sample separately from the first and second diluted samples to allow the formation of an aptamer-affinity complex;
f) The first diluted sample containing the first aptamer affinity complex is transferred to the first mixture, and the first aptamer affinity complex is captured on a solid support in the first mixture. To transfer the above;
g) After step f), the second diluted sample is transferred to the first mixture to form a second mixture, and the second aptamer-affinitive complex of the second diluted solution is the second. The formation, trapped on a solid support in a mixture;
h) The third diluted sample is transferred to the second mixture to form a third mixture, and the third aptamer-affinity complex of the third diluted solution is a solid support in the third mixture. The transfer, captured on;
i) To detect the presence of the first, second, and third aptamers of the first, second, and third aptamer-affinity complexes, or to determine their levels, or 1 Detecting the presence or amount of one or more of the first, second, and third aptamer-affinity complexes;
Including
The first and second diluents are different diluents of the same test sample.
The method.

The method of claim 1, wherein the first and second adapter-target molecule affinity complex is a non-covalent complex.

How to:
a) The first capture reagent is brought into contact with the first diluent to form the first mixture, and the second capture reagent is brought into contact with the second diluent to form the second mixture. Each of the second capture reagents is immobilized on a solid support, and each of the first and second capture reagents has an affinity for a different target molecule, said formation;
b) Incubating the first and second mixtures separately, the first capture reagent-target molecule if the target molecule with which the first capture reagent has an affinity is present in the first mixture. When the affinity complex is formed in the first mixture and the target molecule with which the second capture reagent has an affinity is present in the second mixture, the second capture reagent-target molecule affinity complex is present. The incubation formed in the second mixture;
c) The third capture reagent is brought into contact with the third diluent to form a third mixture, the third capture reagent is immobilized on a solid support, and the third capture reagent is the first capture reagent. And the formation, which has an affinity for a target molecule different from the second capture reagent;
d) (i) The first capture reagent-target molecule affinity complex, followed by the second capture reagent-target molecule affinity complex, and (ii) the second capture reagent-target molecule affinity complex, Subsequently, in the order selected from the first capture reagent-target molecule affinity complex, the first and second capture reagent-target molecule affinity complexes are sequentially released and combined in the fourth mixture. ;
e) Attach the first tag to the target molecule of the first and second capture reagents-target molecule affinity complex;
f) The tagged first and second capture reagents-target molecule affinity complex is contacted with one or more solid supports and the tags are tagged with the first and second capture reagents-target molecule affinity. To immobilize the complex on one or more of the solid supports;
g) Dissociate the capture reagent from the capture reagent-target molecule affinity complex;
h) Detecting the presence of the dissociated capture reagent, or determining its level;
The first and second diluents are different diluents for the test sample.
The method.

The method of claim 3, wherein the first and second capture reagent-target protein affinity complexes are non-covalent complexes.

The method of claim 3, wherein the first and second capture reagents are independently selected from an aptamer or antibody.

The method according to claim 5, wherein the first and second capture reagents are aptamers, respectively.

The third capture, further comprising incubating the third mixture separately from the first and second mixtures, if the target molecule with which the third capture reagent has an affinity is present in the third mixture. The method according to any one of claims 3 to 6, wherein the reagent-target molecule affinity complex is formed in the third mixture.

(I) The first capture reagent-target molecule affinity complex, followed by the second capture reagent-target molecule affinity complex, followed by the third capture reagent-target molecule affinity complex; (ii) said. The first capture reagent-target molecule affinity complex, followed by the third capture reagent-target molecule affinity complex, followed by the second capture reagent-target molecule affinity complex; (iii) the second capture reagent. -Target molecule affinity complex, followed by the third capture reagent-target molecule affinity complex, followed by the first capture reagent-target molecule affinity complex; (iv) the second capture reagent-target molecule affinity. The sex complex, followed by the first capture reagent-target molecule affinity complex, followed by the third capture reagent-target molecule affinity complex; (v) the third capture reagent-target molecule affinity complex, The first capture reagent-target molecule affinity complex, followed by the second capture reagent-target molecule affinity complex; and (vi) the third capture reagent-target molecule affinity complex, followed by the said. The third capture reagent-target molecule affinity complex is released in order selected from the second capture reagent-target molecule affinity complex, and then the first capture reagent-target molecule affinity complex; 7. The method of claim 7, further comprising combining the first and second capture reagents-target molecule affinity complex with the fourth mixture.

How to:
a) The first capture reagent is contacted with the first diluent to form the first mixture, the second capture reagent is contacted with the second diluent to form the second mixture, and the third capture reagent is third diluted. Contacting with the liquid to form a third diluted mixture, each of the first, second and third capture reagents is immobilized on a solid support and the first and second capture reagents are respectively immobilized. And each of the third capture reagents has an affinity for different target molecules, said formation;
b) Incubating the first, second and third mixtures separately, the first capture if the target molecule with which the first capture reagent has an affinity is present in the first mixture. If the reagent-target molecule affinity complex is formed in the first mixture and the target molecule with which the second capture reagent has an affinity is present in the second mixture, the second capture reagent-target molecule affinity. When the sex complex is formed in the second mixture and the target molecule with which the third capture reagent has an affinity is present in the third mixture, the third capture reagent-target molecule affinity complex is said. The incubation formed in the third mixture;
c) (i) The first capture reagent-target molecule affinity complex, followed by the second capture reagent-target molecule affinity complex, followed by the third capture reagent-target molecule affinity complex; (ii). ) The first capture reagent-target molecule affinity complex, followed by the third capture reagent-target molecule affinity complex, followed by the second capture reagent-target molecule affinity complex; (iii) said second. The capture reagent-target molecule affinity complex, followed by the third capture reagent-target molecule affinity complex, followed by the first capture reagent-target molecule affinity complex; (iv) the second capture reagent-target. The molecular affinity complex, followed by the first capture reagent-target molecule affinity complex, followed by the third capture reagent-target molecule affinity complex; (v) the third capture reagent-target molecule affinity complex. The body, followed by the first capture reagent-target molecule affinity complex, followed by the second capture reagent-target molecule affinity complex; and (vi) the third capture reagent-target molecule affinity complex, followed by The affinity complex is released in order selected from the second capture reagent-target molecule affinity complex, and then the first capture reagent-target molecule affinity complex; in the fourth mixture. Combine in;
d) Attach the first tag to the target molecule of the first, second and third capture reagents-target molecule affinity complex;
e) The tagged first, second and third capture reagents-target molecular affinity complex is contacted with one or more solid supports and the tag is attached to the first, second and said. Third capture reagent-to ensure that the target molecule affinity complex is immobilized on one or more of the solid supports;
f) Dissociate the capture reagent from the capture reagent-target molecule affinity complex;
g) Including detecting the presence of the dissociated capture reagent or determining its level;
The first, second and third diluents are different diluents of the test sample.
The method.

The method of claim 9, wherein the first, second and third capture reagent-target protein affinity complexes are non-covalent complexes.

9. The method of claim 9, wherein the first, second and third capture reagents are independently selected from an aptamer or antibody.

11. The method of claim 11, wherein the first, second and third capture reagents are aptamers, respectively.

The test samples include plasma, serum, urine, whole blood, leukocytes, peripheral blood mononuclear cells, buffy coat, sputum, tears, mucus, nasal lavage fluid, nasal aspirate, semen, saliva, peritoneal lavage fluid, ascites, cyst fluid, Claimed to be selected from medullary fluid, sheep's fluid, glandular fluid, lymphatic fluid, papillary aspirate, bronchial aspirate, bronchial scraping fluid, salivary fluid, joint plasma fluid, organ secretions, cells, cell extracts and cerebrospinal fluid. The method according to any one of 1 to 12.

The target molecules are proteins, peptides, carbohydrates, polysaccharides, glycoproteins, hormones, receptors, antigens, antibodies, viruses, bacteria, metabolites, cofactors, inhibitors, drugs, dyes, nutrients, growth factors, cells and tissues. The method according to any one of claims 1 to 13, which is selected from.

Detecting the presence of dissociated first and second capture reagents, or determining their levels, is performed by PCR, mass spectrometry, nucleic acid sequencing, next-generation sequencing (NGS) or hybridization. Item 6. The method according to any one of Items 1 to 14.

The method of any one of claims 1, 2, 5, 6-8 and 11-15, wherein each aptamer independently comprises at least one 5-position modified pyrimidine.

16. The method of claim 16, wherein the at least one 5-position modified pyrimidine comprises a linker at the 5-position of the pyrimidine and a moiety bound to the linker.

17. The method of claim 17, wherein the linker is selected from an amide linker, a carbonyl linker, a propynyl linker, an alkyne linker, an ester linker, a urea linker, a carbamate linker, a guanidine linker, an amidine linker, a sulfoxide linker and a sulfone linker.

17. The method of claim 17 or 18, wherein the moiety is a hydrophobic moiety.

The above part is the following part:

19. The method of claim 19.

Claimed, wherein the moiety is selected from a naphthyl moiety, a benzyl moiety, a fluorobenzyl moiety, a tyrosine moiety, an indole moiety, a morpholino moiety, an isobutyl moiety, a 3,4-methylenedioxybenzyl moiety, a benzothiophenyl moiety and a benzofuranyl moiety. 19. The method according to 19.

The method according to any one of claims 16 to 21, wherein the pyrimidine of the 5-position modified pyrimidine is uridine, cytidine or thymidine.

Claims 5, 6 that the third diluted sample is incubated separately from the first and second diluted samples, allowing the formation of an aptamer-affinity complex between the third aptamer and its target molecule. The method according to any one of 8 and 11-22.

It further comprises transferring the third diluted sample to the second mixture to form a third mixture, wherein the third aptamer-affinitive complex of the third diluted solution is on a solid support in the third mixture. 23. The method of claim 23, which is captured in.

Claimed further comprising detecting the presence or level of the third aptamer of the third aptamer affinity complex, or detecting the presence or amount of the third aptamer affinity complex. Item 24.

The method according to any one of claims 1 to 25, wherein the third diluted solution is a diluted solution different from the first and second diluted solutions of the same test sample.

The first diluted solution is 0.001% to 0.009% (or 0.001%, 0.002%, 0.003%, 0.004%, 0.005%, 0.006%, 0). .007%, 0.008% or 0.009%), or 0.002% to 0.008%, or 0.003% to 0.007%, or about 0.005%, said test. It is a diluted solution of the sample; and the second diluted solution is 0.01% to 1% (or 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, 0.1%, 0.15%, 0.2%, 0.25%, 0.3%, 0.35%, 0.4%, 0.45%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9% or 1%), or 0.1% to 0.8% , Or 0.2% to 0.75%, or about 0.5%, diluted solution of the test sample.
The method according to any one of claims 1 to 26.

The first diluted solution is 0.001% to 0.009% (or 0.001%, 0.002%, 0.003%, 0.004%, 0.005%, 0.006%, 0). .007%, 0.008% or 0.009%), or 0.002% to 0.008%, or 0.003% to 0.007%, or about 0.005%, said test. It is a diluted solution of the sample; and the second diluted solution is 5% to 39% (or 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30% , 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38% or 39%), or 15% to 30%, or 15% to 25%, or about 20%. , Which is a diluted solution of the test sample.
The method according to any one of claims 1 to 26.

The first diluted solution is 0.01% to 1% (or 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07). %, 0.08%, 0.09%, 0.1%, 0.15%, 0.2%, 0.25%, 0.3%, 0.35%, 0.4%, 0.45 %, 0.5%, 0.6%, 0.7%, 0.8%, 0.9% or 1%), or 0.1% -0.8%, or 0.2%- 0.75% or about 0.5% diluted solution of the test sample; and the second diluted solution is 5% to 39% (or 5%, 6%, 7%, 8%). , 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25 %, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38% or 39%), or 15% ~ A diluted solution of 30%, 15% to 25%, or about 20% of the test sample.
The method according to any one of claims 1 to 26.

The first diluted solution is 0.01% to 1% (or 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07). %, 0.08%, 0.09%, 0.1%, 0.15%, 0.2%, 0.25%, 0.3%, 0.35%, 0.4%, 0.45 %, 0.5%, 0.6%, 0.7%, 0.8%, 0.9% or 1%), or 0.1% -0.8%, or 0.2%- 0.75%, or about 0.5%, diluted solution of the test sample; and said second diluted solution is 0.001% to 0.009% (or 0.001%, 0. 002%, 0.003%, 0.004%, 0.005%, 0.006%, 0.007%, 0.008% or 0.009%), or 0.002% to 0.008% , Or 0.003% to 0.007%, or about 0.005%, diluted solution of the test sample.
The method according to any one of claims 1 to 26.

The first diluent is 5% to 39% (or 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%). , 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33. %, 34%, 35%, 36%, 37%, 38% or 39%), or 15% to 30%, or 15% to 25%, or about 20% diluted solution of the test sample. And the second diluent is 0.01% to 1% (or 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%). , 0.07%, 0.08%, 0.09%, 0.1%, 0.15%, 0.2%, 0.25%, 0.3%, 0.35%, 0.4% , 0.45%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9% or 1%), or 0.1% to 0.8%, or 0 .2% to 0.75%, or about 0.5%, diluted solution of the test sample.
The method according to any one of claims 1 to 26.

The first diluent is 5% to 39% (or 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%). , 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33. %, 34%, 35%, 36%, 37%, 38% or 39%), or 15% to 30%, or 15% to 25%, or about 20% diluted solution of the test sample. And the second diluent is 0.001% to 0.009% (or 0.001%, 0.002%, 0.003%, 0.004%, 0.005%, 0. 006%, 0.007%, 0.008% or 0.009%), or 0.002% to 0.008%, or 0.003% to 0.007%, or about 0.005%. , Which is a diluted solution of the test sample.
The method according to any one of claims 1 to 26.

The third diluent is:
5% -39% (or 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19 %, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38% or 39%), 15% -30%, 15% -25%, about 20%;
0.01% to 1% (or 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0 .09%, 0.1%, 0.15%, 0.2%, 0.25%, 0.3%, 0.35%, 0.4%, 0.45%, 0.5%, 0 6.6%, 0.7%, 0.8%, 0.9% or 1%), 0.1% -0.8%, 0.2% -0.75%, about 0.5%; and 0.001% to 0.009% (or 0.001%, 0.002%, 0.003%, 0.004%, 0.005%, 0.006%, 0.007%, 0.008% Or 0.009%), or 0.002% to 0.008%, 0.003% to 0.007%, about 0.005%;
The method according to any one of claims 1 to 32, which is a diluted solution of the test sample selected from.

The first diluted solution is 0.001% to 0.009% (or 0.001%, 0.002%, 0.003%, 0.004%, 0.005%, 0.006%, 0). .007%, 0.008% or 0.009%), or 0.002% to 0.008%, or 0.003% to 0.007%, or about 0.005%, said test. A diluted solution of the sample;
The second diluted solution contains 0.01% to 1% (or 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07). %, 0.08%, 0.09%, 0.1%, 0.15%, 0.2%, 0.25%, 0.3%, 0.35%, 0.4%, 0.45 %, 0.5%, 0.6%, 0.7%, 0.8%, 0.9% or 1%), or 0.1% -0.8%, or 0.2%- 0.75% or about 0.5% diluted solution of the test sample; and the third diluted solution is 5% to 39% (or 5%, 6%, 7%, 8%). , 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25 %, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38% or 39%), or 15% ~ 30%, or 15% to 25%, or about 20% diluted solution of the test sample;
The method according to any one of claims 1 to 26.

The first diluted solution is 0.001% to 0.009% (or 0.001%, 0.002%, 0.003%, 0.004%, 0.005%, 0.006%, 0). .007%, 0.008% or 0.009%), or 0.002% to 0.008%, or 0.003% to 0.007%, or about 0.005%, said test. It is a diluted solution of the sample;
The second diluent is 5% to 39% (or 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%). , 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33. %, 34%, 35%, 36%, 37%, 38% or 39%), or 15% to 30%, or 15% to 25%, or about 20% diluted solution of the test sample. And the third diluent is 0.01% to 1% (or 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%). , 0.07%, 0.08%, 0.09%, 0.1%, 0.15%, 0.2%, 0.25%, 0.3%, 0.35%, 0.4% , 0.45%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9% or 1%), or 0.1% to 0.8%, or 0 .2% to 0.75%, or about 0.5%, diluted solution of the test sample;
The method according to any one of claims 1 to 26.

The first diluted solution is 0.01% to 1% (or 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07). %, 0.08%, 0.09%, 0.1%, 0.15%, 0.2%, 0.25%, 0.3%, 0.35%, 0.4%, 0.45 %, 0.5%, 0.6%, 0.7%, 0.8%, 0.9% or 1%), or 0.1% -0.8%, or 0.2%- 0.75% or about 0.5% diluted solution of the test sample;
The second diluent is 5% to 39% (or 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%). , 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33. %, 34%, 35%, 36%, 37%, 38% or 39%), or 15% to 30%, or 15% to 25%, or about 20% diluted solution of the test sample. And the third diluent is 0.001% to 0.009% (or 0.001%, 0.00).
2%, 0.003%, 0.004%, 0.005%, 0.006%, 0.007%, 0.008% or 0.009%), or 0.002% to 0.008% , Or 0.003% to 0.007%, or about 0.005%, diluted solution of the test sample.
The method according to any one of claims 1 to 26.

The first diluted solution is 0.01% to 1% (or 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07). %, 0.08%, 0.09%, 0.1%, 0.15%, 0.2%, 0.25%, 0.3%, 0.35%, 0.4%, 0.45 %, 0.5%, 0.6%, 0.7%, 0.8%, 0.9% or 1%), or 0.1% -0.8%, or 0.2%- 0.75% or about 0.5% diluted solution of the test sample;
The second diluent is 0.001% to 0.009% (or 0.001%, 0.002%, 0.003%, 0.004%, 0.005%, 0.006%, 0). .007%, 0.008% or 0.009%), or 0.002% to 0.008%, or 0.003% to 0.007%, or about 0.005%, said test. It is a diluted solution of the sample; and the third diluted solution is 5% to 39% (or 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30% , 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38% or 39%), or 15% to 30%, or 15% to 25%, or about 20%. Is a diluted solution of the test sample;
The method according to any one of claims 1 to 26.

The first diluent is 5% to 39% (or 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%). , 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33. %, 34%, 35%, 36%, 37%, 38% or 39%), or 15% to 30%, or 15% to 25%, or about 20% diluted solution of the test sample. And;
The second diluted solution contains 0.01% to 1% (or 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07). %, 0.08%, 0.09%, 0.1%, 0.15%, 0.2%, 0.25%, 0.3%, 0.35%, 0.4%, 0.45 %, 0.5%, 0.6%, 0.7%, 0.8%, 0.9% or 1%), or 0.1% -0.8%, or 0.2%- 0.75% or about 0.5% diluted solution of the test sample.
The third diluted solution contains 0.001% to 0.009% (or 0.001%, 0.002%, 0.003%, 0.004%, 0.005%, 0.006%, 0). .007%, 0.008% or 0.009%), or 0.002% to 0.008%, or 0.003% to 0.007%, or about 0.005%, said test. A diluted solution of the sample;
The method according to any one of claims 1 to 26.

The first diluent is 5% to 39% (or 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%). , 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33. %, 34%, 35%, 36%, 37%, 38% or 39%), or 15% to 30%, or 15% to 25%, or about 20% diluted solution of the test sample. And;
The second diluent is 0.001% to 0.009% (or 0.001%, 0.002%, 0.003%, 0.004%, 0.005%, 0.006%, 0). .007%, 0.
008% or 0.009%), or 0.002% to 0.008%, 0.003% to 0.007%, or about 0.005% diluted solution of the test sample. And the third diluent is 0.01% to 1% (or 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0). .07%, 0.08%, 0.09%, 0.1%, 0.15%, 0.2%, 0.25%, 0.3%, 0.35%, 0.4%, 0 .45%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9% or 1%), or 0.1% -0.8%, or 0.2 % To 0.75%, or about 0.5%, diluted solution of the test sample;
The method according to any one of claims 1 to 26.