Application of K1C13 tear concentration detection reagent in the preparation of pregnancy tear identification reagent

Abstract

The present invention discloses the application of a K1C13 tear concentration detection reagent in the preparation of a pregnancy tear identification reagent, and belongs to the field of pregnancy detection. The present invention finds that compared with normal non-pregnant women, the tear concentration of K1C13 protein in pregnant women in early pregnancy is significantly increased, and has the property of being a pregnancy identification marker. Therefore, the K1C13 tear concentration detection reagent has the application of preparing a pregnancy identification reagent. The present invention provides a new method for the preparation of a pregnancy identification reagent, which is conducive to improving the accuracy and sensitivity of pregnancy detection technology.

Description

Technical Field

[0001] The invention relates to the field of pregnancy detection, and particularly to application of a K1C13 tear concentration detection reagent in the preparation of a pregnancy tear identification reagent. Background Art

[0002] The current pregnancy test only uses human chorionic gonadotropin (hCG) as a single marker, and pregnancy identification is achieved by detecting hCG levels in urine or blood. The identification of more markers and the development of more diverse detection methods can effectively supplement existing methods and promote the development of existing methods in the direction of multiple markers, which is conducive to improving the accuracy and sensitivity of pregnancy detection technology. The identification program generated by artificial intelligence training has gradually replaced the manually generated identification formula or competent experience to become the mainstream tool for identification. The elements of artificial intelligence training include features, feature data, and machine learning or deep learning algorithms suitable for feature data types. Summary of the invention

[0003] In view of this, one of the objects of the present invention is to provide a use of a K1C13 tear concentration detection reagent in the preparation of a pregnancy tear identification reagent.

[0004] In order to achieve the above object, the present invention provides the following technical solutions:

[0005] 1. Application of K1C13 tear concentration detection reagent in the preparation of pregnancy tear identification reagent.

[0006] Preferably, the K1C13 is received as P13646 in the Uniprot database.

[0007] Preferably, in the present invention, the K1C13 tear concentration refers to detecting the concentration of K1C13 protein in the tears of a subject using the tears of the subject as the test sample.

[0008] Preferably, the principle of the K1C13 tear concentration detection reagent is affinity labeling, spectroscopy, chromatography, and mass spectrometry.

[0009] Preferably, the principle of the K1C13 tear concentration detection reagent is enzyme-linked immunosorbent assay.

[0010] Preferably, the pregnancy tear identification reagent of the present invention refers to a reagent capable of distinguishing between non-pregnant and pregnant women by detecting the tears of a subject.

[0011] The beneficial effects of the present invention are as follows: the present invention discloses a new tear identification reagent for pregnancy developed based on a new tear marker, namely K1C13 tear protein, which is an effective supplement to the existing pregnancy detection methods, will promote the development of pregnancy detection technology in the direction of multiple markers, and is conducive to improving the accuracy and sensitivity of pregnancy detection technology. BRIEF DESCRIPTION OF THE DRAWINGS

[0012] In order to make the purpose, technical solution and beneficial effects of the present invention clearer, the present invention provides the following drawings for illustration:

[0013] Figure 1 These are the differentially expressed proteins in the tears of non-pregnant and early pregnant women identified by proteomics technology;

[0014] Figure 2 The results of tear proteomics study show the difference in tear concentration of K1C13 between the non-pregnant and early pregnant groups;

[0015] Figure 3 The ELISA test results showed the difference in tear concentration of K1C13 between the non-pregnant and early pregnancy groups;

[0016] Figure 4 The AUC values ​​for K1C13 tear protein concentrations to identify pregnancy. DETAILED DESCRIPTION

[0017] The present invention is further described below in conjunction with the accompanying drawings and specific embodiments so that those skilled in the art can better understand the present invention and implement it, but the embodiments are not intended to limit the present invention.

[0018] Example 1: Using tear proteomics to study the differences in tear protein profiles between non-pregnant and early pregnant women

[0019] 19 volunteers were recruited from normal women (Group C), normal pregnant women in early pregnancy (Group ES), mid-pregnancy (Group MS), and gestational diabetes (Group GDM). The inclusion criteria were: no history of pregnancy, no history of chronic diseases, no history of medication within 6 months, no medical abnormalities during pregnancy, no history of ophthalmic diseases, and no abnormalities of key organs as shown by physical examination. Tear samples were obtained, and the tear proteome profiles of normal women and pregnant women were studied using a proteomics method based on iTRAQ technology. Differential proteins were found by comparison. All differential proteins were as follows: Figure 1 shown.

[0020] Among the differentially expressed proteins, the concentration of K1C13 tear protein in early pregnancy women was significantly higher than that in non-pregnant women ( Figure 2), and therefore was selected as a candidate tear protein marker. For the tear sample preparation and tear protein profiling method based on iTRAQ (Isobaric tags for relative and absolute quantitation) in this example, please refer to Zhou L, Beuerman RW, Chan CM, Zhao SZ, Li XR, Yang H, Tong L, Liu S, Stern ME, Tan D. Identification of tear fluid biomarkers in dry eye syndrome using iTRAQ quantitative proteomics. J Proteome Res. 2009 Nov; 8(11): 4889-905. doi: 10.1021 / pr900686s. PMID: 19705875. and Liu J, Qiu J, Zhang Z, Zhou L, Li Y, Ding D, Zhang Y, Zou D, Wang D, Zhou Q, Lang T. SOX4 maintains the stemness of cancer cells via transcriptionally enhancing HDAC1 revealed by comparative proteomics study. Cell Biosci.2021Jan 22;11(1):23.doi:10.1186 / s13578-021-00539-y.PMID:33482915; PMCID:PMC7821488.

[0021] Example 2: Identification of K1C13 concentration differences in tears of pregnant women in early pregnancy using ELISA

[0022] 182 normal women (Group C) and 176 normal pregnant women in early pregnancy (Group ES) were recruited as research volunteers. The inclusion criteria were the same as those described in Example 1. Tear samples were obtained and the tear concentration of K1C13 was detected by ELISA. The results showed that the tear protein concentration of K1C13 was significantly high in pregnant women in early pregnancy ( Figure 3). For the ELISA method in this embodiment, see Sakamoto S, Putalun W, Vimolmangkang S, Phoolcharoen W, Shoyama Y, Tanaka H, ​​Morimoto S. Enzyme-linked immunosorbent assay for the quantitative / qualitative analysis of plant secondary metabolites. J Nat Med. 2018 Jan; 72(1): 32-42. doi: 10.1007 / s11418-017-1144-z. Epub 2017 Nov 21. Erratumin: J Nat Med. 2018 Jan 5; PMID: 29164507; PMCID: PMC5775980.

[0023] Example 3: AUC value for identifying pregnancy using K1C13 tear protein concentration

[0024] The AUC value for identifying pregnancy using K1C13 tear protein concentration as an indicator was calculated using the data obtained in Example 2. The results are as follows: Figure 4 As shown, the AUC value was 0.8226, and when the threshold was 2.076 ng / ul, the sensitivity was 72.57% and the specificity was 70.88%.

[0025] The above-described embodiments are only preferred embodiments for fully illustrating the present invention, and the protection scope of the present invention is not limited thereto. Equivalent substitutions or changes made by those skilled in the art based on the present invention are within the protection scope of the present invention. The protection scope of the present invention shall be subject to the claims.

Claims

1. The application of K1C13 tear concentration detection reagent in the preparation of pregnancy tear identification reagent, characterized in that: The K1C13 has a receiving number of P13646 in the Uniprot database; the pregnancy tear identification reagent refers to a reagent that can distinguish between non-pregnant and pregnant women by detecting the tears of the subject.

2. The use according to claim 1, characterized in that: The K1C13 tear concentration refers to the concentration of K1C13 protein in the tears of a subject, which is detected by taking the tears of the subject as the test sample.

3. The use according to claim 1, characterized in that: The principle of the K1C13 tear concentration detection reagent is affinity labeling, spectroscopy, chromatography, and mass spectrometry.

4. The use according to claim 1, characterized in that: The principle of the K1C13 tear concentration detection reagent is enzyme-linked immunosorbent assay.

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