Fluorescent PCR (polymerase chain reaction) nucleic acid detection reagent strip for detecting listeria monocytogenes

Abstract

The utility model relates to the technical field of biological detection, in particular to a fluorescent PCR (Polymerase Chain Reaction) nucleic acid detection reagent strip for detecting Listeria monocytogenes, which is characterized in that after a sample to be detected is added into the reagent strip through a sample adding hole, the sample flows into a nucleic acid extraction area for nucleic acid extraction, and the extracted nucleic acid enters an amplification reaction area for amplification reaction; when a target nucleic acid sequence is amplified and then enters fluorescence detection to generate a fluorescence signal, a sensor in a fluorescence detection area captures the fluorescence signal in real time and transmits the signal to a control chip, the control chip processes and analyzes data, and a result is transmitted to external equipment (such as a computer, a mobile phone and the like) through a data transmission module. The rapid detection on the listeria monocytogenes is realized. The reagent strip disclosed by the utility model integrates the functions of nucleic acid extraction, amplification reaction and detection, is simple and convenient to operate, does not need complex sample pretreatment and professional experimental equipment, and is suitable for on-site rapid detection and basic application.

Description

Technical Field

[0001] This invention relates to the field of biological detection technology, and in particular to a fluorescent PCR nucleic acid detection reagent strip for detecting Listeria monocytogenes. Background Technology

[0002] Listeria monocytogenes, commonly known as Listeria, is an important foodborne pathogen that can cause listeriosis in humans. Infection can easily lead to serious diseases such as sepsis, meningitis, and encephalitis. In pregnant women, infection can also cause adverse pregnancy outcomes such as miscarriage and stillbirth. Therefore, rapid and accurate detection of Listeria monocytogenes is of great significance for food safety monitoring, clinical diagnosis, and disease control. Methods for detecting Listeria monocytogenes mainly include culture methods, immunological methods, and molecular biological methods. While culture methods are the gold standard for bacterial detection, the detection cycle is long, usually requiring 3-7 days, which is insufficient for rapid detection. Immunological methods, such as enzyme-linked immunosorbent assay (ELISA), have the advantages of simple operation and fast detection speed, but suffer from low sensitivity and a high risk of false positives. Molecular biological methods, such as polymerase chain reaction (PCR), have the advantages of high sensitivity and specificity, but require specialized laboratory equipment and technicians, and the operation process is relatively complex.

[0003] In the traditional process of detecting Listeria monocytogenes using fluorescent PCR nucleic acid detection strips, sample collection and detection are often two separate steps. It is necessary to first collect the sample using a special sampling tool, and then transfer the sample to the detection strip or instrument for detection. The operation process is complicated and the detection process is relatively cumbersome, which is not conducive to rapid on-site detection. At the same time, the sample may be lost or contaminated during the sample transfer process, which cannot guarantee the accuracy of the detection.

[0004] Therefore, there is an urgent need for a fluorescent PCR nucleic acid detection reagent strip that can rapidly detect Listeria monocytogenes and is easy to operate, in order to solve the problems mentioned in the background technology. Utility Model Content

[0005] To address the current issues of requiring specialized laboratory equipment and complex procedures for detecting Listeria monocytogenes, which hinder rapid on-site detection and easily lead to sample contamination, a fluorescent PCR nucleic acid detection strip for detecting Listeria monocytogenes has been invented.

[0006] The technical solution of this utility model includes a shell, wherein a fixing plate is provided at the lower end of the shell, a placement cavity is provided on the fixing plate, a chromatography detection strip is provided in the placement cavity, and a nucleic acid extraction area, an amplification reaction area, a fluorescence detection area, and an absorption area are sequentially arranged on the chromatography detection strip. A sample addition hole is provided on the shell for adding the sample to be tested. The sample addition hole is located above the nucleic acid extraction area. A sampling component is provided on the shell for injecting the sample into the sample addition hole.

[0007] Preferably, the housing is provided with a detachable fluorescence detection sensor, which includes a control chip and a data transmission module.

[0008] Preferably, the sampling component includes a rubber air bladder, the outer shell is provided with a rubber air bladder located above the sampling port, the outer shell is provided with a sampling tube, and one end of the sampling tube near the sampling tube is connected to the rubber air bladder.

[0009] Preferably, the sampling tube is configured in a "Z" shape, and a fixing protrusion is provided on the outer shell. The sampling tube is fixed on the fixing protrusion, and the top height of the sampling tube is higher than the top height of the rubber airbag.

[0010] Preferably, the upper end of the outer casing is provided with a display unit located in the fluorescence detection area.

[0011] Preferably, the upper end of the outer shell is provided with an anti-slip part.

[0012] Compared with the prior art, the technical solution of this utility model can achieve the following beneficial effects:

[0013] (1) After the sample to be tested is added to the reagent strip through the sample well, it flows into the nucleic acid extraction area for nucleic acid extraction. The extracted nucleic acid enters the amplification reaction area for amplification. When the target nucleic acid sequence is amplified, it enters the fluorescence detection area to generate a fluorescence signal. The sensor in the fluorescence detection area captures the fluorescence signal in real time and transmits the signal to the control chip. The control chip processes and analyzes the data and transmits the results to external devices (such as computers, mobile phones, etc.) through the data transmission module to realize the rapid detection of Listeria monocytogenes. The reagent strip of this utility model integrates nucleic acid extraction, amplification reaction and detection functions. It is easy to operate and does not require complicated sample pretreatment and professional experimental equipment. It is suitable for rapid on-site detection and grassroots applications.

[0014] (2) A sampling component is set up so that the sample taken from the sampling component can be directly added to the sample well for detection during the sample transfer process, making the sample collection and detection process more coherent and convenient, reducing the loss and contamination during the sample transfer process and ensuring the accuracy of the detection.

[0015] The technical solution of this utility model has broad application prospects in the field of biological detection technology. Attached Figure Description

[0016] Figure 1 This is an isometric view of the present invention.

[0017] Figure 2 This is a top view of the present invention.

[0018] Figure 3 This is a front sectional view of the present invention.

[0019] Figure 4 This is a schematic diagram of the structure of this utility model.

[0020] The components are as follows: 1. Outer shell; 2. Fixing plate; 3. Placement cavity; 4. Chromatography detection strip; 5. Nucleic acid extraction area; 6. Amplification reaction area; 7. Fluorescence detection area; 8. Absorption area; 9. Sample application hole; 10. Sampling component; 11. Fluorescence detection sensor; 12. Rubber airbag; 13. Sampling tube; 14. Fixing protrusion; 15. Display unit; 16. Anti-slip part. Detailed Implementation

[0021] The technical solutions of various embodiments of this utility model will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of this utility model, and not all embodiments. Based on the embodiments of this utility model, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of this utility model. In the description of this utility model, it should be noted that the terms "center", "upper", "lower", "left", "right", "vertical", "horizontal", "inner", "outer", etc., indicate the orientation or positional relationship based on the orientation or positional relationship shown in the accompanying drawings, and are only for the convenience of describing this utility model and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, or be constructed and operated in a specific orientation, and therefore should not be construed as a limitation of this utility model.

[0022] Furthermore, the terms "first," "second," and "third" are used for descriptive purposes only and should not be construed as indicating or implying relative importance. In the description of this utility model, it should be noted that, unless otherwise explicitly specified and limited, the terms "installation," "connection," and "joining" should be interpreted broadly. For example, they can refer to a fixed connection, a detachable connection, or an integral connection; they can refer to a mechanical connection or an electrical connection; they can refer to a direct connection or an indirect connection through an intermediate medium; and they can refer to the internal connection of two components. Those skilled in the art can understand the specific meaning of the above terms in this utility model based on the specific circumstances.

[0023] like Figures 1-4 The fluorescent PCR nucleic acid detection reagent strip for detecting Listeria monocytogenes shown in the illustration includes, in one specific embodiment, a shell 1 with a fixing plate 2 at the lower end of the shell 1. In this embodiment, the shell 1 is a long strip of plastic or paper structure, serving to protect and support the internal structure. The shell 1 and the fixing plate 2 are bonded together. The fixing plate 2 has a placement cavity 3, and a chromatography detection strip 4 is placed inside the placement cavity 3. The shape of the placement cavity 3 is the same as the shape and size of the chromatography detection strip 4, facilitating the fixation of the chromatography detection strip 4. The chromatography detection strip 4 has a nucleic acid extraction zone 5, an amplification reaction zone 6, a fluorescence detection zone 7, and an absorption zone 8 arranged sequentially along the direction of sample flow. In this embodiment, the nucleic acid extraction zone 5 can be made of polyester film, which has good chemical stability, mechanical strength, and water resistance. It is responsible for the uniform diffusion and preliminary filtration of the sample solution, allowing the sample to flow smoothly to the amplification reaction zone 6. As the sample solution flows, it enters the amplification reaction zone 6, which is pre-encapsulated with fluorescent PCR reaction solution, including specific primers, probes, dNTPs, and DNA. In preparing the reagent strips, primers and probes targeting Listeria monocytogenes specific genes are designed. Primers, probes, dNTPs, DNA polymerase, buffer, etc., are prepared according to the formula to form a fluorescent PCR reaction solution, which is then encapsulated in the amplification reaction area 6. When the target nucleic acid sequence is amplified, the probe binds to the amplification product, generating a fluorescent signal. Subsequently, the sample solution enters the fluorescent detection area 7 for detection. The main function of the absorption area 8 is to absorb excess reagents and prevent liquid backflow.

[0024] like Figures 1-4 The fluorescent PCR nucleic acid detection reagent strip for detecting Listeria monocytogenes shown in the figure has a sample application well 9 on the outer shell 1 for adding the sample to be tested. The sample application well 9 is located above the nucleic acid extraction area 5. The outer shell 1 is provided with a sampling component 10 for injecting the sample into the sample application well 9. Through the sampling component 10, the sample obtained in the sampling component 10 can be directly added into the sample application well 9 for detection during the sample transfer process, making the sample collection and detection process more continuous and convenient, reducing loss and contamination during the sample transfer process, and ensuring the accuracy of the detection.

[0025] like Figures 1-4The fluorescent PCR nucleic acid detection reagent strip for detecting Listeria monocytogenes shown in the figure has one specific embodiment in which a detachable fluorescent detection sensor 11 is provided on the outer shell 1. In this embodiment, the fluorescent detection sensor 11 is connected to the upper part of the fluorescent detection area 7 by a snap-fit. The outer shell 1 has a display section 15 located above the fluorescent detection area 7. The fluorescent detection sensor 11 includes a control chip and a data transmission module for controlling the detection process, processing detection data, and transmitting data to external devices. After the fluorescent detection sensor 11 is detached, the reagent strip can also be directly inserted into the detection instrument in the laboratory for detection, which improves the versatility of the device.

[0026] like Figures 1-4 The fluorescent PCR nucleic acid detection reagent strip for detecting Listeria monocytogenes shown in the figure has one specific embodiment, in which the sampling component 10 includes a rubber bladder 12. The rubber bladder 12 is disposed on the outer shell 1 above the sample application hole 9. A sampling capillary 13 is fixedly connected to the outer shell 1. One end of the sampling capillary 13 is connected to the rubber bladder 12. When the rubber bladder 12 is pressed and the sampling capillary 13 is inserted into the sample solution to be sampled, the rubber bladder 12 is released. The sample solution is injected into the rubber bladder 12 through the sampling capillary 13 by the recovery of the rubber bladder 12, and then injected into the sample application hole 9 again for detection.

[0027] like Figures 1-4 The fluorescent PCR nucleic acid detection reagent strip shown is used to detect Listeria monocytogenes. In one specific embodiment, the sampling tube 13 is set in a "Z" shape, and a fixing protrusion 14 is provided on the outer shell 1. The sampling tube 13 is fixed on the fixing protrusion 14. The top height of the sampling tube 13 is higher than the top height of the rubber air bladder 12. After sampling, the reagent strip is laid flat, and the sample solution is automatically injected into the sample well 9 under the action of gravity.

[0028] like Figures 1-4 The fluorescent PCR nucleic acid detection reagent strip shown is for detecting Listeria monocytogenes. In one specific embodiment, the upper end of the outer shell 1 has an anti-slip part 16 for easy hand-holding and use.

[0029] The working principle of this device has been explained through the above embodiments. These embodiments only illustrate several implementation methods of this utility model, and while the descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the utility model patent. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of this utility model, and these all fall within the protection scope of this utility model. Therefore, the protection scope of this utility model patent should be determined by the appended claims.

Claims

1. A fluorescent PCR nucleic acid detection reagent strip for detecting Listeria monocytogenes, comprising a shell (1), characterized in that, The lower end of the outer shell (1) is provided with a fixing plate (2), and a placement cavity (3) is provided on the fixing plate (2). A chromatography detection strip (4) is provided in the placement cavity (3). A nucleic acid extraction area (5), an amplification reaction area (6), a fluorescence detection area (7), and an absorption area (8) are arranged sequentially on the chromatography detection strip (4). A sample loading hole (9) is provided on the outer shell (1) for adding the sample to be tested. The sample loading hole (9) is located above the nucleic acid extraction area (5). A sampling component (10) is provided on the outer shell (1) for injecting the sample into the sample loading hole (9).

2. The fluorescent PCR nucleic acid detection reagent strip for detecting Listeria monocytogenes according to claim 1, characterized in that, The housing (1) is provided with a detachable fluorescence detection sensor (11), which includes a control chip and a data transmission module.

3. The fluorescent PCR nucleic acid detection reagent strip for detecting Listeria monocytogenes according to claim 1, characterized in that, The sampling assembly (10) includes a rubber airbag (12). The outer shell (1) is provided with a rubber airbag (12) located above the sampling hole (9). The outer shell (1) is provided with a sampling tube (13). One end of the sampling tube (13) near the sampling tube (13) is connected to the rubber airbag (12).

4. The fluorescent PCR nucleic acid detection reagent strip for detecting Listeria monocytogenes according to claim 3, characterized in that, The sampling tube (13) is set in a "Z" shape, and a fixing protrusion (14) is provided on the outer shell (1). The sampling tube (13) is fixed on the fixing protrusion (14), and the top height of the sampling tube (13) is higher than the top height of the rubber airbag (12).

5. The fluorescent PCR nucleic acid detection reagent strip for detecting Listeria monocytogenes according to claim 1, characterized in that, The upper end of the outer casing (1) is provided with a display unit (15) located in the fluorescence detection area (7).

6. The fluorescent PCR nucleic acid detection reagent strip for detecting Listeria monocytogenes according to claim 1, characterized in that, The upper end of the outer shell (1) is provided with an anti-slip part (16).

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