A chip for high-throughput nucleic acid mass spectrometry

Abstract

The application discloses a chip for high-throughput nucleic acid mass spectrum detection, which comprises a hydrophilic coating (1) and a hydrophobic substrate (2), wherein the hydrophilic coating (1) is arranged on the surface of the hydrophobic substrate (2); the hydrophilic coating (1) is divided into a plurality of square detection islands (5) by transverse division lines (3) and longitudinal division lines (4), and the thickness of the hydrophilic coating (1) is less than 20 microns; the transverse division lines (3) and the longitudinal division lines (4) are made of hydrophobic materials; and the hydrophilic coating material is a metal oxide. The hydrophilic oxide is used as the coating, the resistance to strong laser irradiation is significantly improved, the chip can be repeatedly used, the detection cost is significantly reduced, and the application range is widened.

Description

Technical Field

[0001] This invention belongs to the field of analytical detection, specifically relating to a chip for high-throughput nucleic acid mass spectrometry detection. Technical Field

[0002] Nucleic acid mass spectrometry combines multiplex nucleic acid amplification, high-throughput chips, and matrix-assisted laser desorption / ionization time-of-flight mass spectrometry (MALDI-TOF), enabling the simultaneous detection of dozens of targets on a single sample. It can process over three thousand samples per day, and the results are simple to analyze, requiring no professional interpretation. Nucleic acid mass spectrometry overcomes the drawbacks of current gene sequencing methods, such as multiple steps, long processing times, and high costs, while also compensating for the low throughput and accuracy of quantitative real-time PCR. It holds great promise in molecular diagnostics fields such as food safety, genetic disease screening, pathogen detection, and genomics.

[0003] High-throughput nucleic acid chips are a key component. Currently, there are very few nucleic acid chips suitable for nucleic acid mass spectrometry, and patented technologies remain largely controlled by foreign companies. The steel target plates produced by Shimadzu and Bruker lack chemical modification and have poor crystal morphology, resulting in low accuracy, low signal-to-noise ratio, and high baseline for nucleic acid mass spectrometry peak detection. Agena has developed the SpectroCHIP chip with 96 and 384 wells for nucleic acid mass spectrometry. Through surface treatment and matrix development, it avoids sample aggregation on the high-density solid-state chip during sample loading, improving crystal uniformity. However, it is expensive, and patents and matrix formulations are monopolized by foreign companies. Internationally, Lawrence Livermore National Laboratory in the United States has been dedicated to introducing advanced processes such as photolithography and precision surface micro / nano fabrication, attempting to optimize and improve chip structure and materials to enhance chip stability and sensitivity. In China, Ma Qingwei et al. invented a universal chip for time-of-flight mass spectrometry detection of proteins and nucleic acids (CN201710539893.0). The chip in this invention includes: (1) a chip body: the surface of the body has a micro-arranged hydrophilic spotting well and a hydrophobic outer area; (2) a chip adapter made of a metal substrate, wherein the surface of the adapter is distributed with multiple chip slots that match the chip body. The chip body is selected from a chip made of diamond, single-crystal silicon, or quartz crystal that is tough and has good flatness. The hydrophilic spotting well is covered with a hydrophilic film of 150-800 nm, and the hydrophobic outer area is covered with a hydrophobic film of 150 nm-2 μm with a water droplet contact angle >120°. The spotting well is pretreated with a matrix solution. The matrix solution is selected from a matrix solution used for protein spectrometry or a matrix solution used for nucleic acid mass spectrometry. Bu Junfa et al. disclosed a mass spectrometry chip and its preparation method and application (CN202111253001.3). The mass spectrometry chip includes: a substrate; a film coating formed on the surface of the substrate, the film coating being made of micro / nano materials; and a metal and metal oxide layer formed on the surface of the film coating. This invention constructs a novel mass spectrometry chip with micro / nano materials as the film layer and surface sputtering of plasmon noble metals, enhancing the detection efficiency of laser desorption / ionization mass spectrometry for small molecule metabolites. The detection time is short, with results obtained in as little as 1 minute; the detection throughput is high, enabling large-scale, high-throughput biological sample detection; and it can distinguish the metabolic profiles of different populations based on serum samples, laying a theoretical foundation for early disease screening and diagnosis. The aforementioned patent uses organic film materials as the coating, which are easily damaged under laser irradiation and cannot be used for long periods. Summary of the Invention

[0004] The purpose of this invention is to provide a high-throughput nucleic acid mass spectrometry detection chip. A high-temperature resistant hydrophilic metal oxide is used as the hydrophilic coating; the hydrophilicity allows the sample to remain within the detection island, while the high-temperature resistance enhances its ability for long-term repeated use.

[0005] To achieve the above objectives, the technical solution adopted by the present invention is as follows:

[0006] The present invention provides a chip for high-throughput nucleic acid mass spectrometry detection, comprising a hydrophilic coating (1) and a hydrophobic substrate (2), wherein the hydrophilic coating (1) is located on the surface of the hydrophobic substrate (2);

[0007] The hydrophilic coating (1) is divided into several square detection islands (5) by transverse dividing lines (3) and longitudinal dividing lines (4), and the thickness of the hydrophilic coating (1) is less than 20 micrometers;

[0008] The horizontal dividing line (3) and the vertical dividing line (4) are made of hydrophobic material;

[0009] The hydrophilic coating material is a metal oxide.

[0010] The chip has X and Y arrows marking a corner, and the detection islands are named according to the coordinate axes, such as (Xa, Yb) being the detection island in row a and column b.

[0011] Furthermore, in the above technical solution, the metal oxide is selected from one or a mixture of several of titanium dioxide, manganese oxide, copper oxide, iron oxide, aluminum oxide, tungsten oxide, and zinc oxide.

[0012] Furthermore, in the above technical solution, the hydrophobic substrate material is selected from quartz, glass, silicon wafer, and stainless steel.

[0013] Furthermore, in the above technical solution, the transverse dividing line (3) and the longitudinal dividing line (4) are the exposed hydrophobic substrate material after the metal oxides on the corresponding positions of the hydrophilic coating (1) are peeled off.

[0014] Furthermore, in the above technical solution, the horizontal dividing line (3) and the vertical dividing line (4) are hydrophobic organic compound coatings.

[0015] Furthermore, in the above technical solution, the hydrophobic organic compound coating is selected from polytetrafluoroethylene (PTFE), fluorinated polyethylene, siloxane compounds, stearic acid compounds, organometallic framework compounds (MOFs), vitrimers, perfluorinated compounds, or fluorinated surfactants.

[0016] Furthermore, the number of detection islands on a single chip can be 96 or 384.

[0017] Furthermore, in the above technical solution, the side length of the detection island (5) is between 1 mm and 10 mm.

[0018] This invention provides a chip for high-throughput nucleic acid mass spectrometry detection that can be used in conjunction with laser-assisted desorption / time-of-flight mass spectrometry, which helps to reduce detection costs and broaden application scenarios.

[0019] The advantages of this invention are: when hydrophilic oxides are used as coatings, the resistance to strong laser irradiation is significantly improved, reusability is achieved, detection costs are significantly reduced, and the application range is broadened. Attached Figure Description

[0020] Figure 1 This is a schematic diagram of the chip structure for high-throughput nucleic acid mass spectrometry detection according to the present invention;

[0021] In the figure, 1. Hydrophilic coating; 2. Hydrophobic substrate; 3. Horizontal dividing line; 4. Vertical dividing line; 5. Square detection island; 6. X-axis; 7. Y-axis. Detailed Implementation

[0022] Example 1

[0023] like Figure 1 As shown, a chip for high-throughput nucleic acid mass spectrometry detection includes a hydrophilic coating 1 and a hydrophobic substrate 2, wherein the hydrophilic coating 1 is located on the surface of the hydrophobic substrate 2;

[0024] The hydrophilic coating 1 is divided into several square detection islands 5 by transverse dividing lines 3 and longitudinal dividing lines 4, and the thickness of the hydrophilic coating 1 is 10 micrometers.

[0025] The horizontal dividing line 3 and the vertical dividing line 4 are made of hydrophobic material;

[0026] The hydrophilic coating material is a metal oxide. The metal oxide is selected from titanium dioxide.

[0027] The detection islands are named according to the coordinate axes, with X and Y arrows marking a corner of the chip. For example, Xa and Yb are the detection islands in row a and column b, respectively.

[0028] The hydrophobic substrate material is selected from quartz.

[0029] The horizontal dividing line 3 and the vertical dividing line 4 are the exposed hydrophobic substrate material after the metal oxides on the corresponding positions of the hydrophilic coating 1 are peeled off.

[0030] The side length of detection island 5 is 5 millimeters.

[0031] The number of detection islands on a single chip can be 96.

[0032] The above-mentioned high-throughput nucleic acid mass spectrometry detection chip is combined with laser-assisted desorption / time-of-flight mass spectrometry, which helps to reduce detection costs and broaden application scenarios.

[0033] Example 2

[0034] like Figure 1 As shown, a chip for high-throughput nucleic acid mass spectrometry detection includes a hydrophilic coating 1 and a hydrophobic substrate 2, wherein the hydrophilic coating 1 is located on the surface of the hydrophobic substrate 2;

[0035] The hydrophilic coating 1 is divided into several square detection islands 5 by transverse dividing lines 3 and longitudinal dividing lines 4, and the thickness of the hydrophilic coating 1 is 5 micrometers.

[0036] The horizontal dividing line 3 and the vertical dividing line 4 are made of hydrophobic material;

[0037] The hydrophilic coating material is a metal oxide. The metal oxide is selected from copper oxide.

[0038] The detection islands are named according to the coordinate axes, with X and Y arrows marking a corner of the chip. For example, Xa and Yb are the detection islands in row a and column b, respectively.

[0039] The hydrophobic substrate material is selected from stainless steel.

[0040] The horizontal dividing line 3 and the vertical dividing line 4 are hydrophobic organic compound coatings. The hydrophobic organic compound coatings are metal-organic frameworks (MOFs).

[0041] The side length of detection island 5 is 2 millimeters.

[0042] The number of detection islands on a single chip can be up to 384.

[0043] The above-mentioned high-throughput nucleic acid mass spectrometry detection chip is combined with laser-assisted desorption / time-of-flight mass spectrometry, which helps to reduce detection costs and broaden application scenarios.

Claims

1. A chip for high-throughput nucleic acid mass spectrometry detection, characterized in that: It includes a hydrophilic coating (1) and a hydrophobic substrate (2), with the hydrophilic coating (1) located on the surface of the hydrophobic substrate (2); The hydrophilic coating (1) is divided into several square detection islands (5) by transverse dividing lines (3) and longitudinal dividing lines (4), and the thickness of the hydrophilic coating (1) is less than 20 micrometers; The horizontal dividing line (3) and the vertical dividing line (4) are made of hydrophobic material; The hydrophilic coating material is a metal oxide.

2. The chip for high-throughput nucleic acid mass spectrometry detection according to claim 1, characterized in that: The metal oxide is selected from one or a mixture of several of titanium dioxide, manganese oxide, copper oxide, iron oxide, aluminum oxide, tungsten oxide, and zinc oxide.

3. The chip for high-throughput nucleic acid mass spectrometry detection according to claim 1, characterized in that: The hydrophobic substrate material is selected from quartz, glass, silicon wafers, and stainless steel.

4. The chip for high-throughput nucleic acid mass spectrometry detection according to claim 1, characterized in that: The transverse dividing line (3) and the longitudinal dividing line (4) are the exposed hydrophobic substrate material after the metal oxides at the corresponding positions of the hydrophilic coating (1) are peeled off.

5. The chip for high-throughput nucleic acid mass spectrometry detection according to claim 1, characterized in that: The horizontal dividing line (3) and the vertical dividing line (4) are hydrophobic organic compound coatings.

6. The chip for high-throughput nucleic acid mass spectrometry detection according to claim 5, characterized in that: The hydrophobic organic compound coating is selected from polytetrafluoroethylene, fluorinated polyethylene, siloxane compounds, stearic acid compounds, organometallic framework compounds, glass polymers, perfluorinated compounds, or fluorinated surfactants.

7. The chip for high-throughput nucleic acid mass spectrometry detection according to claim 1, characterized in that: The side length of the detection island (5) is between 1 mm and 10 mm.

8. The chip for high-throughput nucleic acid mass spectrometry detection according to claim 1, characterized in that: The number of detection islands on a single chip is 96 or 384.

9. A high-throughput nucleic acid mass spectrometry detection chip according to any one of claims 1-7, when used in conjunction with laser-assisted desorption / time-of-flight mass spectrometry, helps to reduce detection costs and broaden application scenarios.

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