Jettisonable kit cooperating with portable Raman spectrometer for detection

Abstract

The invention relates to a jettisonable kit cooperating with a portable Raman spectrometer for detection. The jettisonable kit comprises a kit body, and the top surface of the kit body is provided with a group of SERS scanning holes, a pull rod hole and a sample adding port; a pull rod is arranged in the pull rod hole; a sample tube is arranged in the box body; a group of capturing devices are arranged in the box body, and each capturing device is in transmission connection with the pull rod through a transmission mechanism; each capturing device is provided with a vertically arranged capillary tube bundle, and the tail end of each capillary tube bundle extends into the sample tube; a group of SERS (Surface Enhanced Raman Scattering) detection substrates adopting micro-fluidic chips are arranged above the capturing device in the box body; each SERS detection substrate is positioned right below the corresponding SERS scanning hole; and a base protection device for locking the pull rod is arranged in the box body. According to the kit disclosed by the invention, the SERS technology and the microfluidic technology are utilized, and a plurality of SERS detection substrates can be used at the same time, so that a plurality of indexes of the same sample can be rapidly detected, and the kit can be applied to rapid detection of sudden cardiac death risk assessment.

Description

Technical Field

[0001] The invention relates to a disposable test kit for coordinated detection with a portable Raman spectrometer, belonging to the SERS technical field. Background Art

[0002] Raman scattering spectrum is a molecular vibration spectrum. Changes in the lattice and bonding of the material will cause the position of the characteristic spectral band or peak to shift. Surface-enhanced Raman spectroscopy (SERS) is a method that increases the surface roughness of precious metals and adsorbs the object to be tested onto the metal surface, making the scattered laser intensity 10 times stronger than ordinary Raman. 6 -10 11 The method has the characteristics of high sensitivity and short detection time. It can be directly used for non-destructive testing or in-situ analysis of analytical materials without complicated pretreatment.

[0003] Microfluidics refers to the science and technology involved in systems that use microchannels (tens to hundreds of microns in size) to process or manipulate tiny fluids (with volumes ranging from picoliters to nanoliters). It is an emerging interdisciplinary subject involving chemistry, fluid physics, microelectronics, new materials, biology, and biomedical engineering.

[0004] At present, the "gold standard" for diagnosing myocardial ischemia is coronary angiography; coronary angiography is an invasive procedure that may cause embolic complications both during and after the procedure. It is also expensive and may impose a greater financial burden on patients. High-performance liquid chromatography can also be used, but this method takes a long time to detect, requires a high-throughput developer for pre-separation, and can only measure one marker at a time. It has high requirements for technical personnel and is complex to operate, making it impossible to complete rapid on-site detection.

[0005] Therefore, it is urgent to develop technical means to rapidly detect multiple markers using SERS technology and microfluidics technology. Summary of the invention

[0006] The main purpose of the present invention is to overcome the problems existing in the prior art and provide a disposable kit for coordinated detection with a portable Raman spectrometer, which can use multiple SERS detection substrates at the same time, thereby realizing rapid detection of multiple indicators of the same sample, and can be applied to rapid detection of sudden cardiac death risk assessment, and can be used immediately after detection.

[0007] The technical solution of the present invention to solve the technical problem is as follows:

[0008] A disposable test kit for coordinated detection with a portable Raman spectrometer comprises a box body, wherein a group of SERS scanning holes, a pull rod hole and a sample addition port are arranged on the top surface of the box body; a pull rod is arranged in the pull rod hole; a sample tube is arranged in the box body, one end of the sample tube is connected to the sample addition port, and the other end is connected to the side wall of the box body; a group of capture devices are arranged in the box body, each capture device is connected to the pull rod through a transmission mechanism; each capture device has a vertically arranged capillary bundle, and the end of each capillary bundle extends into the sample tube; a group of SERS detection substrates using a microfluidic chip are arranged above the capture device in the box body, each SERS detection substrate corresponds to the capture device one by one, each SERS detection substrate corresponds to the SERS scanning hole one by one, and each SERS detection substrate is located directly below the corresponding SERS scanning hole; a substrate protection device for locking the pull rod is arranged in the box body.

[0009] The reagent kit comprises: a substrate protection device locking rod, and an initial state in which each SERS detection substrate and a corresponding capture device are separated from each other.

[0010] The reagent kit also has: the substrate protection device is disengaged from the pull rod to unlock it, the pull rod is pulled out of the box body under the action of external force, and the pull rod moves each capture device to a processing state combined with the corresponding SERS detection substrate through a transmission mechanism.

[0011] In this structure, in the initial state, the locking rod of the substrate protection device can ensure that the SERS detection substrate and the capture device of the reagent kit are kept separated from each other before actual use, thereby ensuring its usability; in actual use, after adding the test sample, the reagent kit is placed in a processing state. At this time, the capture device and the SERS detection substrate are combined to form a sandwich structure, and a portable Raman spectrometer can be used to align the SERS scanning hole to detect each SERS detection substrate to obtain the test result.

[0012] The technical solution further improved by the present invention is as follows:

[0013] Preferably, the sample tube is formed by connecting a vertical tube and a horizontal tube, the top of the vertical tube is connected with a sample adding port, and the end of the horizontal tube away from the vertical tube is connected to the side wall of the box body.

[0014] More preferably, the sample tube is in an "L" shape.

[0015] By adopting the above preferred solutions, the specific structure of the sample tube can be further optimized.

[0016] Preferably, the substrate protection device has a laterally arranged rod-shaped member, the rod-shaped member is provided with a limiting protrusion near the pull rod, and the upper part of the pull rod is provided with a limiting notch; when the reagent box is in the initial state, the limiting protrusion of the rod-shaped member is engaged with the limiting notch of the pull rod.

[0017] A connecting seat is fixedly connected to the lower part of the pull rod, and the connecting seat has an external thread; a limiting sleeve is fixedly provided in the box body in the circumferential direction of the pull rod, and the limiting sleeve has an internal thread; the external thread of the connecting seat matches the internal thread of the limiting sleeve; a connecting piece is coaxially provided at the bottom of the connecting seat, and the connecting piece and the connecting seat constitute a rotating pair with axial constraint, wherein the connecting seat is the mover and the connecting piece is the stator; a cross bar is fixedly connected to the lower end of the connecting piece, and each capture device is installed on the cross bar; the connecting seat, the limiting sleeve, the connecting piece and the cross bar together constitute a transmission mechanism.

[0018] More preferably, the process of the test kit entering the processing state from the initial state is: the rod-shaped member of the substrate protection device is pulled outward so that the limiting protrusion of the rod-shaped member and the limiting notch of the pull rod are separated from each other; the pull rod is pulled out of the box body under the action of external force, and when the connecting seat contacts the limiting sleeve, the pull rod is rotated to thread the connecting seat and the limiting sleeve until each capture device moves to combine with the corresponding SERS detection substrate.

[0019] By adopting the above preferred solutions, the specific structures of the substrate protection device and the transmission mechanism can be further optimized.

[0020] Preferably, the sample loading port is provided with a sealing device, which includes a protective film, a buffer strip, a spring and a sealing strip; there are two buffer strips and they are symmetrically distributed on both sides of the central axis of the sample loading port and are parallel to each other, the buffer strips are respectively sealed and fixedly connected to the tube wall of the sample loading port, a gap for loading is reserved between the two buffer strips, and the sealing strip is respectively bonded to the two buffer strips and seals and covers the gap; there are two protective films and they correspond to the buffer strips one by one, and the area between the side of each buffer strip away from the sealing strip and the tube wall of the sample loading port is respectively sealed and covered by the corresponding protective film, one side of the protective film is sealed and fixedly connected to the tube wall of the sample loading port and the other side is sealed and fixedly connected to the corresponding buffer strip; there are two springs and they correspond to the buffer strips one by one, the spring expansion and contraction direction is perpendicular to the extension direction of the buffer strip, one end of the spring is fixedly connected to the tube wall of the sample loading port and the other end is fixedly connected to the corresponding buffer strip; a vacuum area is provided on the side wall of the box body, and the sample tube is connected to the vacuum area.

[0021] By adopting the above preferred solution, the specific structure of the reagent kit can be further optimized. In this way, when in use, the sealing strip of the sample adding port is torn off, and then the blood sample is added, and the blood sample can be delivered into the sample tube and contact each capture device faster and more smoothly by means of the suction airflow formed in the vacuum area.

[0022] Preferably, the box body is made of plastic.

[0023] After adopting the above preferred scheme, for disposable test kits, in terms of material selection, plastic products are low in price, easy to make, and relatively stable, and should be the first choice, which can fully utilize the practicality and portability of plastic products.

[0024] Preferably, the kit is an SMF sudden cardiac death early screening kit; there are six SERS scanning holes, six SERS detection substrates, and six specific capture devices, and they correspond one to one; each SERS detection substrate is: a high-sensitivity troponin T detection substrate, a creatine kinase isoenzyme detection substrate, a aspartate aminotransferase detection substrate, a lactate dehydrogenase B detection substrate, a pyruvate detection substrate, and a spermine detection substrate.

[0025] More preferably, the method of using the kit is as follows: add the sample to be tested to the sample addition port, the sample to be tested enters the sample tube, contacts the capillary bundle of each capture device and enters therein; operate the substrate protection device to unlock the pull rod, and use the pull rod to move each capture device to combine with the corresponding SERS detection substrate to form a sandwich structure; then, use a portable Raman spectrometer to detect each SERS detection substrate through the SERS scanning hole, and obtain the detection result.

[0026] More preferably, each SERS detection substrate is obtained by the following preparation process:

[0027] The silicon wafer was immersed in a piranha solution in an alkaline tank for 2 hours, washed with deionized water, then immersed in the piranha solution for 2 hours, and then washed with deionized water; the silicon wafer was cultured in a solution containing a specific antibody of 33mM PDA for 1 hour to obtain a PDA chip; the PDA chip was washed with deionized water and stored at 4°C for later use; the piranha solution was prepared by mixing 98% concentrated sulfuric acid and 30% hydrogen peroxide solution in a ratio of 7:3; the specific antibody was one of a high-sensitivity troponin T antibody, a creatine kinase isoenzyme antibody, a aspartate aminotransferase antibody, a lactate dehydrogenase B antibody, a pyruvate antibody, and a spermine antibody; each PDA chip was respectively loaded into a microfluidic chip to obtain each SERS detection substrate.

[0028] The kit of the present invention utilizes SERS technology and microfluidic technology, and can use multiple SERS detection substrates at the same time, thereby realizing rapid detection of multiple indicators of the same sample, and can be applied to rapid detection of sudden cardiac death risk assessment. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 This is a schematic diagram of the overall structure of Example 1 of the present invention.

[0030] Figure 2 This is a schematic diagram of a top view of Embodiment 1 of the present invention.

[0031] Figure 3 Schematic diagram of the transmission mechanism of embodiment 1 of the present invention.

[0032] Figure 4 Schematic diagram of the sample adding port of Example 1 of the present invention.

[0033] Figures 5 to 10They are respectively the test result diagrams of Example 3 of the present invention. Among them, Figure 5 This is the test result of high-sensitivity troponin T. Figure 6 This is the test result of creatine kinase isoenzyme. Figure 7 This is the test result of aspartate aminotransferase. Figure 8 This is the test result of pyruvic acid. Fig. 9 This is the test result of lactate dehydrogenase B. Fig.10 This is the detection result of spermine.

[0034] Fig.11 This is a Raman spectrum diagram obtained from 40 blood samples in Example 3 of the present invention. DETAILED DESCRIPTION

[0035] The present invention will be described in further detail below with reference to the accompanying drawings and in combination with embodiments, but the present invention is not limited to the examples given.

[0036] Example 1

[0037] like Figure 1 to Figure 2 As shown, the disposable test kit for coordinated portable Raman spectrometer detection of this embodiment includes a box body 01, the top surface of the box body 01 is provided with a group of SERS scanning holes 02, a pull rod hole 03 and a sample injection port 04; a pull rod 05 is provided in the pull rod hole 03; a sample tube 06 is provided in the box body 01, one end of the sample tube 06 is connected to the sample injection port 04, and the other end is connected to the side wall of the box body; a group of capture devices 07 are provided in the box body 01, and each capture device 07 is connected to the pull rod 05 through a transmission mechanism; each capture device 07 has The capillary bundles are arranged vertically, and the ends of each capillary bundle extend into the sample tube 06 respectively; a group of SERS detection substrates 08 using microfluidic chips are provided above the capture device 07 in the box body 01, and each SERS detection substrate 08 corresponds one-to-one to the capture device 07, and each SERS detection substrate 08 corresponds one-to-one to the SERS scanning hole 02, and each SERS detection substrate 08 is located directly below the corresponding SERS scanning hole 02; a substrate protection device 09 for locking the pull rod 05 is provided in the box body 01.

[0038] The specific conditions of the kit of this embodiment are:

[0039] (1) The substrate protection device 09 locks the pull rod 05, and each SERS detection substrate 08 and the corresponding capture device 07 are separated from each other in the initial state.

[0040] (2) The substrate protection device 09 is disengaged from the pull rod 05 to unlock it, and the pull rod 05 is pulled out of the box body 01 under the action of external force. The pull rod 05 moves each capture device 07 to a processing state combined with the corresponding SERS detection substrate 08 through the transmission mechanism. In this processing state, the capture device 07 and the SERS detection substrate 08 are combined to form a sandwich structure, and at this time, the portable Raman spectrometer can be used to align the SERS scanning hole 02 to detect each SERS detection substrate 08.

[0041] Specifically, the sample tube 06 is formed by connecting a vertical tube and a horizontal tube, the top of the vertical tube is connected to the sample port 04, and the end of the horizontal tube away from the vertical tube is connected to the side wall of the box body. The sample tube 06 is "L" shaped.

[0042] like Figure 3 As shown, the base protection device 09 has a laterally arranged rod-shaped member 10, the rod-shaped member 10 is provided with a limiting protrusion 11 near the pull rod 05, and the upper part of the pull rod 05 is provided with a limiting notch 12; when the test kit is in the initial state, the limiting protrusion 11 of the rod-shaped member 10 is engaged with the limiting notch 12 of the pull rod 05.

[0043] A connecting seat 13 is fixedly connected to the lower part of the pull rod 05, and the connecting seat 13 has an external thread; a limiting sleeve 14 is fixedly provided circumferentially on the pull rod 05 inside the box body 01, and the limiting sleeve 14 has an internal thread; the external thread of the connecting seat 13 matches the internal thread of the limiting sleeve 14; a connecting piece 15 is coaxially provided at the bottom of the connecting seat 13, and the connecting piece 15 and the connecting seat 13 constitute a rotating pair with axial constraint, wherein the connecting seat 13 is a mover, and the connecting piece 15 is a stator; the lower end of the connecting piece 15 is fixedly connected to a cross bar 16, and each capture device 07 is installed on the cross bar 16; the connecting seat 13, the limiting sleeve 14, the connecting piece 15, and the cross bar 16 together constitute a transmission mechanism.

[0044] The process of the reagent kit entering the processing state from the initial state is as follows: the rod-shaped member 10 of the substrate protection device 09 is pulled outward, so that the limiting protrusion 11 of the rod-shaped member 10 and the limiting notch 12 of the pull rod 05 are separated from each other; the pull rod 05 is pulled out of the box body 01 under the action of external force, and when the connecting seat 13 contacts the limiting sleeve 14, the pull rod 05 is rotated to make the connecting seat 13 and the limiting sleeve 14 threadedly connected until each capture device 07 moves to combine with the corresponding SERS detection substrate 08.

[0045] like Figure 4As shown, the sample loading port 04 is provided with a sealing device, which includes a protective film 17, a buffer strip 18, a spring 19 and a sealing strip 20; there are two buffer strips 18 and they are symmetrically distributed on both sides of the central axis of the sample loading port 04 and are parallel to each other, the buffer strips 18 are respectively sealed and fixedly connected to the tube wall of the sample loading port 04, a gap for loading is reserved between the two buffer strips 18, and the sealing strip 20 is respectively bonded to the two buffer strips 18 and seals and covers the gap; there are two protective films 17 and they correspond to the buffer strips 18 one by one, and the area between the side of each buffer strip 18 away from the sealing strip 20 and the tube wall of the sample loading port 04 is respectively sealed and covered by the corresponding protective film 17, one side of the protective film 17 is sealed and fixedly connected to the tube wall of the sample loading port 04 and the other side is sealed and fixedly connected to the corresponding buffer strip 18; there are two springs 19 and they correspond to the buffer strips 18 one by one, the expansion and contraction direction of the spring 19 is perpendicular to the extension direction of the buffer strip 18, one end of the spring 19 is fixedly connected to the tube wall of the sample loading port 04 and the other end is fixedly connected to the corresponding buffer strip 18.

[0046] At the same time, a vacuum area is provided on the side wall of the box body, and the sample tube 06 is connected to the vacuum area. In this way, when in use, the sealing strip 20 of the sample adding port 04 is torn off, and then a blood sample is added, and the blood sample can be delivered into the sample tube 06 and contact each capture device 07 faster and more smoothly by means of the suction airflow formed by the vacuum area.

[0047] In addition, in this embodiment, the material of the box body 01 is plastic; the size of the box body 01 is 8 cm×2 cm×2 cm; vacuum sealing packaging can be used during storage and transportation to extend the duration of the vacuum zone on the side wall of the box body.

[0048] Example 2

[0049] This embodiment is an SMF sudden cardiac death early screening kit based on Example 1.

[0050] In this embodiment, there are six SERS scanning holes, six SERS detection substrates, and six specific capture devices, and they correspond one to one; the SERS detection substrates are: high-sensitivity troponin T detection substrate, creatine kinase isoenzyme detection substrate, aspartate aminotransferase detection substrate, lactate dehydrogenase B detection substrate, pyruvate detection substrate, and spermine detection substrate.

[0051] The method of using the kit of this embodiment is as follows: add the sample to be tested to the sample adding port, the sample to be tested enters the sample tube, contacts and enters the capillary bundle of each capture device; operate the substrate protection device to unlock the pull rod, use the pull rod to move each capture device to combine with the corresponding SERS detection substrate to form a sandwich structure; then, use a portable Raman spectrometer to detect each SERS detection substrate through the SERS scanning hole, and obtain the test results. Afterwards, analyze and judge according to the test results. If the characteristic peak meets the preset value, it can be highly suspected that the corresponding patient of the sample to be tested has a higher risk of sudden cardiac death.

[0052] Each SERS detection substrate is obtained by the following preparation process:

[0053] The silicon wafer was immersed in a piranha solution in an alkaline tank for 2 hours, washed with deionized water, then immersed in the piranha solution for 2 hours, and then washed with deionized water; the silicon wafer was cultured in a solution containing a specific antibody of 33mM PDA for 1 hour to obtain a PDA chip; the PDA chip was washed with deionized water and stored at 4°C for later use; the piranha solution was prepared by mixing 98% concentrated sulfuric acid and 30% hydrogen peroxide solution in a ratio of 7:3; the specific antibody was one of a high-sensitivity troponin T antibody, a creatine kinase isoenzyme antibody, a aspartate aminotransferase antibody, a lactate dehydrogenase B antibody, a pyruvate antibody, and a spermine antibody; each PDA chip was respectively loaded into a microfluidic chip to obtain each SERS detection substrate.

[0054] Example 3

[0055] This embodiment is a specific application of Embodiment 2.

[0056] Early screening of myocardial ischemia in sudden cardiac death is crucial.

[0057] At present, the number of sudden cardiac death is gradually increasing. The methods for screening include coronary angiography, high performance liquid chromatography, etc., which have the problems of large trauma and difficult operation. The present invention utilizes SERS technology combined with microfluidics technology to provide a low-cost, low-difficulty, fast, high-sensitivity, and minimally invasive detection technology for early screening of sudden cardiac death.

[0058] (I) Testing the sensitivity and repeatability of the kit of Example 2:

[0059] The results of testing different concentrations of markers: standard solutions of high-sensitivity troponin T, creatine kinase isoenzyme, aspartate aminotransferase, lactate dehydrogenase B, pyruvate, and spermine were as follows: Figures 5 to 10 As shown. The Raman spectra of the standard solutions each have an obvious characteristic peak, while the blank control has no characteristic peak. In the range of 1 to 5000 ng / mL concentration of each marker, the SERS signal intensity gradually increases with the increase of concentration. The Raman signal intensity is piecewise positively correlated with the logarithm of the concentration of each marker. The results of further curve fitting show that the kit of Example 2 has a high sensitivity, and there is a good linear relationship between the SERS signal intensity and the logarithm of the concentration of each marker, which provides application prospects for the quantitative analysis of each marker in actual clinical blood samples.

[0060] (II) Testing the specificity of the kit of Example 2:

[0061] Other molecules that may exist in clinical blood samples were selected for detection. After comparison, it was found that the signal intensity detected at the characteristic peak of the above six marker groups was significantly higher than that of other molecule groups, which indicates that the detection ability of the kit of Example 2 for the above six markers is significantly higher than that of other antigens.

[0062] The above results show that the kit of Example 2 has good specificity and selectivity, further showing its excellent performance and can be used for actual clinical analysis. Currently, this field has a large market and high economic benefits.

[0063] (III) The test kit of Example 2 was used to test the blood of 40 patients with myocardial ischemia, and each sample was tested at least 3 times. After the blood sample was dripped into the sample injection port, the blood sample flowed into the sample tube, contacted with and entered the capillary bundles of each capture device, and the substrate protection device was operated to unlock the pull rod, and each capture device was moved to combine with the corresponding SERS detection substrate using the pull rod to form a sandwich structure, and then tested with a portable Raman spectrometer. The final Raman spectrum is as follows: Fig.11 shown.

[0064] On the one hand, the characteristic peaks of the markers can be successfully detected using a portable Raman spectrometer, which can further prove that the blood can smoothly enter the capillary bundles of each capture device and then successfully combine with the SERS detection substrate.

[0065] On the other hand, the characteristic peaks of myocardial ischemia markers (high-sensitivity troponin T, creatine kinase isoenzymes, aspartate aminotransferase, lactate dehydrogenase B, pyruvic acid, spermine) were selected as markers, and the concentrations of 40 blood samples were calculated according to the Raman spectra. The standard content of each marker was then compared, for example, myocardial enzyme spectrum: CK 405.3U / L, CK-MB 196.94U / L, LDH896.65U / L, AST 91.9U / L; troponin 0.238ng / mL. By comparison, the degree of myocardial ischemia in each patient can be obtained: if it is within the normal range, although the patient has symptoms of myocardial ischemia, the risk of sudden cardiac death is still low, and continued maintenance treatment can be considered. If it exceeds the normal range, it can be highly suspected that the patient has a high risk of sudden cardiac death, and active means should be used to treat the patient. In this way, the kit of Example 2 can be used to quickly screen for sudden cardiac death and assist in the discovery of heart disease.

[0066] The base of the kit of the present invention has a good reinforcing structure and reinforcing effect, and has obvious characteristic peaks. According to experimental clinical tests, the success rate of early screening of sudden cardiac death using the kit of the present invention is as high as 99%.

[0067] In addition to the above embodiments, the present invention may also have other implementation modes. Any technical solution formed by equivalent replacement or equivalent transformation falls within the protection scope required by the present invention.

Claims

1. A disposable test kit for use with a portable Raman spectrometer, comprising a box body, characterized in that: The top surface of the box body is provided with a pull rod hole, a sample injection port and a group of SERS scanning holes; a pull rod is provided in the pull rod hole; a sample tube is provided in the box body, one end of the sample tube is connected to the sample injection port, and the other end is connected to the side wall of the box body; a group of capture devices are provided in the box body, and each capture device is connected to the pull rod through a transmission mechanism; each capture device has a vertically arranged capillary bundle, and the end of each capillary bundle extends into the sample tube; a group of SERS detection substrates using microfluidic chips are provided above the capture device in the box body, each SERS detection substrate corresponds to the capture device one by one, each SERS detection substrate corresponds to the SERS scanning hole one by one, and each SERS detection substrate is located directly below the corresponding SERS scanning hole; The box body is provided with a base protection device for locking the pull rod; The kit comprises: the substrate protection device locks the pull rod, and each SERS detection substrate and the corresponding capture device are separated from each other in an initial state; The reagent kit also has: the substrate protection device is disengaged from the pull rod to unlock it, the pull rod is pulled out of the box body under the action of external force, and the pull rod moves each capture device to a processing state combined with the corresponding SERS detection substrate through a transmission mechanism.

2. A disposable kit for use with a portable Raman spectrometer according to claim 1, characterized in that: The sample tube is formed by connecting a vertical tube and a horizontal tube. The top of the vertical tube is connected with a sample adding port, and the end of the horizontal tube away from the vertical tube is connected with the side wall of the box body.

3. A disposable kit for use with a portable Raman spectrometer according to claim 2, characterized in that: The sample tube is in an "L" shape.

4. A disposable kit for use with a portable Raman spectrometer according to claim 1, characterized in that: The substrate protection device comprises a rod-shaped member arranged transversely, wherein the rod-shaped member is provided with a position-limiting protrusion near the pull rod, and the upper part of the pull rod is provided with a position-limiting notch; when the reagent box is in the initial state, the position-limiting protrusion of the rod-shaped member is engaged with the position-limiting notch of the pull rod; The lower part of the pull rod is fixedly connected to a connecting seat, and the connecting seat has an external thread; a limiting sleeve is fixedly provided in the box body in the circumferential direction of the pull rod, and the limiting sleeve has an internal thread; the external thread of the connecting seat matches the internal thread of the limiting sleeve; a connecting piece is coaxially provided at the bottom of the connecting seat, and the connecting piece and the connecting seat constitute a rotating pair with axial constraint, wherein the connecting seat is a mover and the connecting piece is a stator; the lower end of the connecting piece is fixedly connected to a cross bar, and each capture device is installed on the cross bar; the connecting seat, the limiting sleeve, the connecting piece and the cross bar together constitute a transmission mechanism.

5. A disposable kit for use with a portable Raman spectrometer according to claim 4, characterized in that: The process of the test kit entering the processing state from the initial state is as follows: the rod-shaped member of the substrate protection device is pulled outward so that the limiting protrusion of the rod-shaped member and the limiting notch of the pull rod are separated from each other; the pull rod is pulled out of the box body under the action of external force, and when the connecting seat contacts the limiting sleeve, the pull rod is rotated to make the connecting seat and the limiting sleeve threadedly connected until each capture device moves to combine with the corresponding SERS detection substrate.

6. A disposable kit for use with a portable Raman spectrometer according to claim 1, characterized in that: The sample loading port is provided with a sealing device, which includes a protective film, a buffer strip, a spring and a sealing strip; there are two buffer strips, which are symmetrically distributed on both sides of the central axis of the sample loading port and are parallel to each other, the buffer strips are respectively sealed and fixedly connected to the tube wall of the sample loading port, a gap for loading is reserved between the two buffer strips, and the sealing strip is respectively bonded to the two buffer strips and seals and covers the gap; there are two protective films, which correspond to the buffer strips one by one, and the area between the side of each buffer strip away from the sealing strip and the tube wall of the sample loading port is respectively sealed and covered by the corresponding protective film, one side of the protective film is sealed and fixedly connected to the tube wall of the sample loading port and the other side is sealed and fixedly connected to the corresponding buffer strip; there are two springs, which correspond to the buffer strips one by one, the expansion and contraction direction of the springs is perpendicular to the extension direction of the buffer strips, one end of the spring is fixedly connected to the tube wall of the sample loading port and the other end is fixedly connected to the corresponding buffer strip; the side wall of the box body is provided with a vacuum area, and the sample tube is connected to the vacuum area.

7. A disposable kit for use with a portable Raman spectrometer according to any one of claims 1 to 6, characterized in that: The box body is made of plastic.

8. A disposable kit for use with a portable Raman spectrometer according to any one of claims 1 to 6, characterized in that: The kit is an SMF sudden cardiac death early screening kit; there are six SERS scanning holes, six SERS detection substrates, and six specific capture devices, and they correspond to each other; the SERS detection substrates are: a high-sensitivity troponin T detection substrate, a creatine kinase isoenzyme detection substrate, a aspartate aminotransferase detection substrate, a lactate dehydrogenase B detection substrate, a pyruvate detection substrate, and a spermine detection substrate.

9. A disposable kit for use with a portable Raman spectrometer according to claim 8, characterized in that: The method of using the kit is as follows: adding a sample to be tested to the sample adding port, the sample to be tested enters the sample tube, contacts the capillary bundles of each capture device and enters therein; The substrate protection device is operated to unlock the pull rod, and each capture device is moved to combine with the corresponding SERS detection substrate using the pull rod to form a sandwich structure; then, each SERS detection substrate is detected through the SERS scanning hole using a portable Raman spectrometer to obtain the detection results.

10. A disposable kit for use with a portable Raman spectrometer according to claim 8, characterized in that: Each SERS detection substrate is obtained by the following preparation process: The silicon wafer was immersed in a piranha solution in an alkaline tank for 2 hours, washed with deionized water, then immersed in the piranha solution for 2 hours, and then washed with deionized water; the silicon wafer was cultured in a solution containing a specific antibody of 33mM PDA for 1 hour to obtain a PDA chip; the PDA chip was washed with deionized water and stored at 4°C for later use; the piranha solution was prepared by mixing 98% concentrated sulfuric acid and 30% hydrogen peroxide solution in a ratio of 7:3; the specific antibody was one of a high-sensitivity troponin T antibody, a creatine kinase isoenzyme antibody, a aspartate aminotransferase antibody, a lactate dehydrogenase B antibody, a pyruvate antibody, and a spermine antibody; each PDA chip was respectively loaded into a microfluidic chip to obtain each SERS detection substrate.

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