Use of a product in the preparation of a pregnancy tear fluid differential kit

Abstract

The application discloses the biological field and concretely relates to an application of a product in preparation of a pregnancy tear liquid identification kit. Research shows that the LV403 tear liquid concentration of a female is obviously higher than that of a non-pregnant female in early pregnancy. Therefore, the LV403 tear liquid concentration detection reagent can be applied to preparation of a pregnancy tear liquid identification kit. The application provides a new pregnancy detection scheme and is committed to promoting the development of a pregnancy detection technology.

Description

Technical Field

[0001] This invention relates to the field of detection, and specifically to the application of a product in the preparation of a pregnancy tear identification kit. Background Technology

[0002] Current pregnancy testing technologies are based on the level of human chorionic gonadotropin (hCG) in blood or urine, and hCG has long been used as a diagnostic indicator for early pregnancy. With advancements in testing technology, hCG-related tests are becoming increasingly important for diagnosis; however, their accuracy remains limited in early pregnancy or certain special circumstances. Developing more novel biomarker-based testing methods will help advance pregnancy testing technology, enabling more accurate and earlier determination of pregnancy status.

[0003] Secondly, using tears as a sample source offers significant advantages and convenience. Compared to traditional methods of collecting blood and urine samples, obtaining tear specimens is simple, rapid, safe, and non-invasive. Women can easily perform the test at home, replacing frequent trips to medical facilities for sample collection. Furthermore, it allows for earlier detection of pregnancy status, enabling timely intervention and supportive measures. Therefore, tear-based pregnancy testing methods have broad application prospects in the future, providing more options for pregnancy detection and improving their sensitivity, convenience, and accuracy. Summary of the Invention

[0004] Given the limitations of current pregnancy testing methods, this invention aims to provide the application of LV403 tear concentration detection reagents in the preparation of pregnancy tear identification reagents, thus promoting the development of pregnancy testing technology towards greater accuracy, earlier detection, and greater convenience. Research shows that the concentration of LV403 in the tears of women in early pregnancy is significantly higher than that of non-pregnant women; therefore, the LV403 tear concentration detection reagent has the application in the preparation of pregnancy tear identification reagents.

[0005] To achieve the above objectives, the present invention provides the following technical solution:

[0006] The application of a product in the preparation of a pregnancy tear identification kit, characterized in that: the product is a reagent for detecting the concentration of LV403 in tears.

[0007] As a preferred technical solution of the present invention, the LV403 has a receiving number of A0A075B6K6 in the Uniprot database.

[0008] As a preferred technical solution of the present invention, the tear concentration specifically refers to the concentration of LV403 protein in the subject's tears.

[0009] As a preferred technical solution of the present invention, the principle of preparing the tear protein concentration detection reagent specifically includes affinity labeling, spectroscopy, chromatography, and mass spectrometry.

[0010] As a preferred technical solution of the present invention, the principle of preparing the tear protein concentration detection reagent is enzyme-linked immunosorbent assay (ELISA).

[0011] As a preferred technical solution of the present invention, the tear identification reagent can distinguish between non-pregnant and pregnant women by detecting the concentration of target proteins in the tear fluid of the subject.

[0012] The beneficial effects of this invention are as follows: In this invention, based on the principles of affinity labeling, spectroscopy, chromatography, and mass spectrometry, a new pregnancy test kit is developed targeting the differential proteins in the tears of women in early pregnancy and those in non-pregnant women. This is conducive to the multi-directional development of pregnancy testing and to improving the diversity, accuracy, and convenience of testing the tears of women in early pregnancy. Attached Figure Description

[0013] To make the objectives, technical solutions, and beneficial effects of this invention clearer, the following figures are provided for illustration:

[0014] Figure 1 To identify differentially expressed proteins in the tears of non-pregnant and early-pregnant women using proteomics techniques;

[0015] Figure 2 The differentially expressed protein LV403, screened through tear proteomics studies, showed a concentration difference in tears between the non-pregnant and early pregnancy groups.

[0016] Figure 3 The results of ELISA testing showed that LV403 concentrations differed between the non-pregnant and early-pregnant groups.

[0017] Figure 4 The AUC value was used to identify pregnancy by the concentration of LV403 tear protein. Detailed Implementation

[0018] The present invention will be further described below with reference to the accompanying drawings and specific embodiments, so that those skilled in the art can better understand and implement the present invention. However, the embodiments described are not intended to limit the present invention.

[0019] Example 1: Studying the differences in tear protein profiles between non-pregnant and early-pregnant women using tear proteomics technology.

[0020] This study used tear proteomics to investigate the differences in tear protein profiles between non-pregnant and pregnant women. Four groups of volunteers were recruited: normal women (Group C), normal pregnant women in early pregnancy (Group ES), women in mid-pregnancy (Group MS), and women with gestational diabetes mellitus (GDM), with 19 volunteers in each group. Inclusion criteria included: no history of pregnancy, no history of chronic diseases, no medication use within the past 6 months, no medical abnormalities during pregnancy, no history of eye diseases, and no abnormalities in critical organs as shown in a medical examination. Tear samples were collected from different groups, and iTRAQ-based proteomics methods were used to analyze the tear proteomic profiles of normal and pregnant women, identifying differentially expressed proteins. All differentially expressed proteins were obtained as follows: Figure 1 As shown.

[0021] Analysis of the differentially expressed proteins revealed a significant increase in LV403 tear protein concentration in normal pregnant women during early pregnancy compared to non-pregnant women. Figure 2 Therefore, we selected LV403 as a candidate tear protein biomarker. The full name of the LV403 protein is Immunoglobulin lambda variable 4-3, Uniprot ID: A0A075B6K6; gene symbol: IGLV4-3; gene full name is immunoglobulin lambda variable 4-3, Gene ID: 28786.

[0022] For details regarding the tear sample preparation method and the tear protein mapping method based on iTRAQ (Isobaric tags for relative and absolute quantitation) in this embodiment, please refer to the following two references:

[0023] Zhou L, Beuerman RW, Chan CM, Zhao SZ, Li XR, Yang H, Tong L, Liu S, SternME, Tan D. Identification of tear fluid biomarkers in dry eye syndrome using iTRAQ quantitative proteomics.JProteome Res.2009Nov;8(11):4889-905.doi:10.1021 / pr900686s.PMID:19705875.

[0024] 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 transcriptionallyenhancing HDAC1 revealed by comparative proteomics study.Cell Biosci.2021Jan22;11(1):23.doi:10.1186 / s13578-021-00539-y.PMID:33482915;PMCID:PMC7821488.

[0025] Example 2: Identifying the difference in tear concentration of LV403 in pregnant women in early pregnancy using ELISA.

[0026] 182 healthy women (Group C) and 176 pregnant women in early pregnancy (Group ES) were recruited as volunteers, with inclusion criteria as described in Example 1. The concentration of LV403 in the collected tear samples was detected using ELISA. The results indicated that LV403 protein was significantly highly expressed in the tears of pregnant women in early pregnancy. Figure 3 For details regarding the ELISA method in this embodiment, please refer to the following literature: Sakamoto S, Putalun W, Vimolmangkang S, Pholcharoen 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.

[0027] Example 3: Identifying the AUC value of pregnancy by measuring the protein concentration of LV403 in tears.

[0028] Based on the results obtained in Example 2, the AUC value for identifying pregnancy using the protein concentration of LV403 in tears as an indicator was 0.8226. The results are as follows... Figure 4 As shown, when the threshold is set to 6.262 ng / ul, the sensitivity is 68.18% and the specificity is 67.03%.

[0029] The above-described embodiments are merely preferred embodiments provided to fully illustrate the present invention, and the scope of protection of the present invention is not limited thereto. Equivalent substitutions or modifications made by those skilled in the art based on the present invention are all within the scope of protection of the present invention. The scope of protection of the present invention is defined by the claims.

Claims

1. The application of a product in the preparation of a pregnancy tear identification kit, characterized in that: The product is a reagent for detecting the concentration of LV403 in tears; the LV403 has the accession number A0A075B6K6 in the Uniprot database; this tear identification reagent can distinguish between non-pregnant and pregnant women by detecting the concentration of target proteins in the tear fluid of the subject.

2. The application of the product according to claim 1 in the preparation of a pregnancy tear identification kit, characterized in that: The LV403 concentration in tears specifically refers to the concentration of LV403 protein in the subject's tears.

3. The application of the product according to claim 1 in the preparation of a pregnancy tear identification kit, characterized in that: The specific principles for preparing this tear protein concentration detection reagent include affinity labeling, spectroscopy, chromatography, and mass spectrometry.

4. The application of the product according to claim 1 in the preparation of a pregnancy tear identification kit, characterized in that: The principle of preparing this tear protein concentration detection reagent is enzyme-linked immunosorbent assay (ELISA).

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