WO2017088214A1

WO2017088214A1 - Fluorescence-based biological detection system

Info

Publication number: WO2017088214A1
Application number: PCT/CN2015/097332
Authority: WO
Inventors: 王周平; 吴世嘉; 戴邵亮; 段诺; 李琪; 夏雨; 马小媛
Original assignee: 江南大学
Priority date: 2015-11-24
Filing date: 2015-12-15
Publication date: 2017-06-01
Also published as: CN105424662A

Abstract

A fluorescence-based biological detection system, comprising a light emitting module (1), a fluorescence receiving module (2), and a data processing module (3). A cuvette (4) is provided between the light emitting module (1) and the fluorescence receiving module (2), and a detection test solution marked by up-converting phosphor is contained in the cuvette (4). The light emitting module (1) comprises an infrared excitation light source; an excitation light beam emitted from the infrared excitation light source irradiates the cuvette (4) containing the detection test solution by means of an excitation light path; fluorescence emitted by the detection test solution is received by the fluorescence receiving module (2) by means of a receiving light path; the excitation light path and the receiving light path are arranged linearly or at right angles to each other. The fluorescence receiving module (2) is a light intensity sensor, and the output end of the light intensity sensor is connected to the data processing module (3). Real-time quantitative detection on the concentration of target bacteria to be detected in multiple detected solutions can be achieved on the basis of an up-converting phosphor technology, the whole detection system does not need immunochromatography test paper, and the excitation light path and the receiving light path are easier and more convenient to configure.

Description

Title of Invention: Fluorescent Bioassay System

Technical field

[0001] The present invention relates to the field of biological detection technology, and particularly relates to a biological detection system for performing quantitative detection of a specific target bacteria concentration in a plurality of test liquids based on an up-conversion fluorescent labeling technique.

Background technique

[0002] Upconversion fluorescence technology is a labeling technique based on upconversion of fluorescent materials. The so-called up-conversion means that long-wavelength radiation is converted into short-wavelength radiation by Up-Converting Phosphor (UCP), which is composed of several rare earth elements in the crystal lattice of some crystals. Its main components include the main matrix, the absorber, and the emitter. Under the excitation of infrared light, the UCP emits visible light with a wavelength much shorter than that of the excitation light. UCP is prepared into nano-sized particles, labeled with biomolecules, and emits visible fluorescence under the excitation of long-wavelength light source. According to the presence or absence and intensity of fluorescence, the properties and content of the tested biomolecules can be judged.

[0003] The biodetection technology using UCP as a marker can perform non-destructive detection on the test object, and has the advantages of flexible detection, high sensitivity, safe use, and the like, and the UCP marker can be combined with the instrument to the target test object. Perform multiple quantitative tests. In the prior art, there has been a detection system using a UCP marker combined with immunochromatographic test strip technology, which uses an excitation light source to illuminate a test strip with a plurality of functional bands, and is received by an imaging system and an image receiver. The fluorescent image on the paper strip is analyzed and processed by the image processing system. The detection system has the following defects: First, it is necessary to design a test strip of a specific structure, and the test strip includes a plurality of functional strips, which are required Processing signals collected by multiple function belts, the test strip structure is complicated, and the preparation is cumbersome. Second, since the test strip is non-transparent, the excitation light path between the excitation light source and the test strip is passed to the test strip and the image receiver. Between the receiving light path, there must be a refraction angle between the two optical paths with the test strip as the turning point, and in order to be able to illuminate the multiple functional bands of the test strip, there is a need for a specific mathematical relationship between the two angles, so that the excitation The setting of the optical path and the receiving optical path is limited;

Using immunochromatographic test strips, the combination of UCP and bioactive molecules and the test substance are immobilized on the surface of the solid phase carrier by immunoreaction, and a special solid phase carrier is required. Fourth, the test strip is for one-time use. Product, can not be reused.

technical problem [0004] The present applicant has improved the above-mentioned shortcomings in the prior art, and provides a fluorescent biological detection system capable of realizing quantitative determination of the concentration of a target bacteria to be detected in a plurality of tested liquids based on up-conversion fluorescence technology. Detection, the entire detection system does not require immunochromatographic test strips, and the solution for exciting the optical path and receiving optical path is easier.

Technical solution

[0005] The technical solution of the present invention is as follows:

[0006] A fluorescent biological detection system includes a light emitting module, a fluorescent receiving module, and a data processing module, and a cuvette is disposed between the light emitting module and the fluorescent receiving module, and the detecting test of the upconverting fluorescent material mark is contained in the cuvette The illuminating module includes an infrared excitation light source, and the excitation light beam emitted by the infrared excitation light source is irradiated onto the cuvette containing the detection liquid solution through the excitation light path, and the fluorescence emitted by the detection test solution is fluorescently circulated through the receiving light. The receiving module receives, the excitation optical path is arranged in a straight line or a right angle with the receiving optical path; the fluorescent receiving module is a light intensity sensor, and an output end of the light intensity sensor is connected to the data processing module.

[0007] A further technical solution thereof is:

[0008] The detection test solution is composed of an up-conversion fluorescent probe and a pathogen-specific aptamer conjugate, a magnetic probe and a pathogen-specific aptamer conjugate, and a pathogen to be tested.

[0009] The up-converting fluorescent probe is obtained by subjecting an up-converting fluorescent material to amination and avidinization, and the up-converting fluorescent material is NaY0.78F4: Yb0.2, Er0.02 nanoparticle.

[0010] The magnetic probe is an avidinized magnetic bead, the magnetic bead is hydrated high-iron hexahydrate as an iron source, and 1,6-hexanediamine is used as an amino functionalizing agent by a hydrothermal_solvent thermal method synthesis.

[0011] The data processing module includes an amplifying circuit module, an anti-interference module, a single chip signal processing module, and a liquid crystal display module, wherein the amplifying circuit module is configured to amplify the received fluorescent signal, and the anti-interference module is configured to use the light intensity sensor The output voltage signal is converted into a current signal transmission, and is converted into a voltage signal at the line terminal, and the single-chip signal processing module performs data operation processing, and the result is displayed through the liquid crystal display module.

[0012] The data processing module includes a communication module, and the communication module is used for remote communication between the single chip computer and the upper computer. [0013] The infrared excitation light source uses 980 nm near-infrared light.

Advantageous effects of the invention

Beneficial effect

[0014] Technical effects of the present invention:

[0015] Compared with the existing detection system using immunochromatographic test strips, the present invention uses a cuvette containing test solution to perform liquid phase spectrum detection through a cuvette, on the one hand, because of the cuvette For transparent, and the cuvette does not have multiple functional bands of immunochromatographic test strips, the requirements for illumination are required. Compared with the limitation of the specific angle between the laser light path and the receiving optical path in the detection system of the conventional non-transparent test strip, In the invention, the infrared light excitation light path and the fluorescence receiving light path are no longer limited by optical detection. In the detection system of the present invention, the excitation light path and the receiving light path are set by 180° straight line or right angle, in the same optical Under the premise of detection function, the setting of infrared excitation light source, cuvette and fluorescence receiving module in the whole detection system is more convenient. Second, the existing immunochromatographic test paper detection system needs to design test paper with specific structure. Strip, the test strip includes a plurality of functional strips, and it is required to process signals collected by a plurality of functional strips, and the test strip has a complicated structure and is cumbersome to prepare. The UCP conjugate and the test substance are immobilized on the surface of the solid phase carrier by an immunological reaction, and a special solid phase carrier is required. The present invention eliminates the complicated structure and cumbersome immunochromatographic test paper by using the cuvette, and the entire detection system has a simple structure. Third, the traditional immunochromatographic test strip is a one-time use product, and the repeated test requires continuous replacement and installation of a new test strip, and the present invention uses a cuvette, after the contrast dish is cleaned, Reuse.

[0016] Compared with the traditional plate colony counting method, the detection system of the present invention is more rapid, reliable, and stable in quantitative detection of the concentration of the target bacteria to be tested in the tested liquid.

Brief description of the drawing

DRAWINGS

1 is a schematic diagram of the principle of Embodiment 1 of the present invention.

2 is a schematic diagram of the principle of Embodiment 2 of the present invention.

[0019] FIG. 3 is a standard regression curve of fluorescence intensity-concentration of Salmonella detection.

4 is a standard regression curve of fluorescence intensity-concentration for detection of Staphylococcus aureus.

[0021] wherein: 1. a light-emitting module; 2. a fluorescent receiving module; 3. a data processing module; 4. a cuvette. Embodiments of the invention

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS [0022] Specific embodiments of the present invention will be described below with reference to the accompanying drawings.

[0023] Referring to FIG. 1 and FIG. 2, the present invention includes a light emitting module 1, a fluorescent receiving module 2, and a data processing module 3. A cuvette 4 is disposed between the light emitting module 1 and the fluorescent receiving module 2, and the cuvette 4 is disposed. a detection test solution containing an up-converting fluorescent material mark; the light-emitting module 1 includes an infrared excitation light source, and the excitation light beam emitted from the infrared excitation light source is irradiated to the cuvette 4 containing the detection test solution through an excitation light path, The fluorescence emitted by the test solution is received by the receiving light route fluorescent receiving module 2, and the excitation light path is disposed at a right angle or a right angle to the receiving optical path; the fluorescent receiving module 2 is a light intensity sensor, and the output end of the light intensity sensor is connected to the data. Processing module 3.

[0024] Specifically, the data processing module 3 includes an amplifying circuit module, an anti-interference module, a single-chip signal processing module, and a liquid crystal display module, where the amplifying circuit module is configured to amplify the received fluorescent signal, and the anti-interference module is used for The voltage signal outputted by the light intensity sensor is converted into a current signal transmission, and is converted into a voltage signal output at the line terminal, and the single-chip signal processing module performs data operation processing, and the result is displayed through the liquid crystal display module.

[0025] Further, the data processing module 3 includes a communication module, and the communication module is used for remote communication between the single chip microcomputer and the host computer.

[0026] detecting 吋, placing the cuvette 4 containing the detection test solution marked with the up-converting fluorescent material, and exciting by the infrared light source, the detection liquid in the cuvette 4 is a function of the concentration of the test bacteria to be tested. Fluorescence of the relationship, the fluorescence is detected by the fluorescence receiving module 2 to be converted into an electrical signal, and then the amplification process is performed by the amplifying circuit module, and the line interference is considered, and the amplified electrical signal passes through the digital anti-interference module. Transmitting, in conjunction with the A/D conversion of the light intensity sensor itself, the converted digital signal enters the signal processing module of the single chip microcomputer, performs function mapping, arithmetic, and logic processing of the data, and the result of processing and outputting by the single chip microcomputer is The liquid crystal display module actually displays the concentration of the fungus to be tested. Further, the present invention has a special RS232 interface, which can be connected with the 32-bit host computer, and can remotely monitor the data result through the remote communication technology. Terminal data analysis.

[0027] The present invention uses a cuvette 4 to hold a test solution labeled with an up-converting fluorescent material, and the present invention only needs to prepare an up-converting fluorescent probe, a magnetic probe, and a suspension of the test bacteria to be tested, up-conversion fluorescence detection. Needle, magnetic The probes were mixed and incubated at room temperature to form a sandwich sandwich complex, magnetically separated from the unbound up-conversion fluorescent probe, and the supernatant was decanted, and BB was used for precipitation (10 mM Tris-HCl, pH 7.4, 100 mM). After the KClfPlmM MgCl ₂ ) buffer is resuspended, the resuspended sandwich sandwich test solution is placed in the cuvette 4, and the infrared light is excited by the infrared light source to contrast the color dish 4, and the sandwich sandwich test is performed. The liquid is subjected to fluorescence intensity measurement to draw a fluorescence intensity-concentration standard regression curve of the pathogen to be tested, and the standard regression curve is set in the detection system of the present invention, and the target bacteria concentration of the sample of unknown concentration can be measured, and the same Test results were verified with the results of the classical plate colony counting method. Compared with the existing detection system using immunochromatographic test strips, the present invention uses a cuvette containing test solution to perform liquid chromatography detection through a cuvette, on the one hand, since the cuvette is transparent And the cuvette does not have the requirement that the plurality of functional bands of the immunochromatographic test paper require illumination, and the infrared in the present invention is different from the limitation condition of the specific angle between the laser light path and the receiving optical path in the detection system of the conventional non-transparent test strip. The optical excitation light path and the fluorescence receiving light path are no longer limited by optical detection. According to the square shape of the cuvette, in the detection system of the present invention, the excitation light path and the receiving light path are 180° linearly arranged or At right angles, the detection system described in Figure 1 is 180 for the excitation and reception paths. The detection system shown in FIG. 2 is arranged at a right angle to the excitation optical path and the receiving optical path, preferably 180° linearly arranged, so that the setting of the infrared excitation light source, the cuvette and the fluorescence receiving module in the entire detection system is simpler; Second, the existing immunochromatographic test strip detection system needs to design a test strip of a specific structure. The test strip includes a plurality of functional strips, and needs to process signals collected by a plurality of functional strips. The test strip has a complicated structure and is cumbersome to prepare. Moreover, the UCP conjugate and the test substance are immobilized on the surface of the solid phase carrier by an immune reaction, and a special solid phase carrier is required. The present invention eliminates the complicated structure and cumbersome immunochromatographic test paper by using the cuvette, and the entire detection system structure Simple, simpler to make; Third, the traditional immunochromatographic test strip is a one-time use product, repeated testing requires constant replacement and installation of new test strips, and the present invention uses a cuvette, after the contrasting dish is cleaned, Can be reused.

[0028] The infrared excitation light source of the invention adopts 980 nm near-infrared excitation light, which is photochemically stable, has strong environmental adaptability, strong penetrating power and no damage, and has relatively low cost.

[0029] Specifically, the detection test solution is composed of an up-conversion fluorescent probe and a pathogen-specific aptamer conjugate, a magnetic probe and a pathogen-specific aptamer conjugate, and a pathogen to be tested. The up-converting fluorescent probe is obtained by amination and avidinization of an up-converting fluorescent material, and the up-converting fluorescent material is NaY ₀ , ₈ F ₄ : Yb 0.2, Er. . . . ₂ nanoparticle; the magnetic probe is an avidinized magnetic bead, the magnetic bead is ferrous chloride hexahydrate as an iron source, and 1,6-hexanediamine is used as an amino functionalizing agent by hydrothermal-solvent heat Method synthesis. Specific preparation of the up-converting fluorescent probe and the magnetic probe will be described below.

[0030] The preparation process of the up-converting fluorescent probe is as follows:

[0031] In the first step, NaY o. ₇₈ F ₄ : Yb was synthesized by a hydrothermal-solvent method. ₂ , Er. . . . ₂ upconverting nanoparticles

(hereinafter referred to as UCNPs): Take Y ₂ 0 ₃ , Yb ₂ 0 ₃ , Er ₂ 0 ₃ (Ln=Y:Yb:Er=78:0.2:0.02), add the three mixtures to an appropriate amount of nitric acid to dissolve, and Volatile excess nitric acid is evaporated to obtain a rare earth nitrate powder; the rare earth nitrate powder is dissolved in 8 mL of deionized water, and 2.1273 g of EDTA (ethylenediaminetetraacetic acid is added, and its molar ratio to RE ³ + is 1 :1) and adjust the pH to weakly alkaline to form a clear and transparent EDTA-Ln ³⁺ solution; take 25 mL of ethylene glycol, force into 0.4 g CTAB (cetyltrimethylammonium bromide) and the above EDTA-Ln 3+ solution, about 3 mL of HF (hydrofluoric acid) was added dropwise with rapid stirring to obtain a white milky colloid. Finally, 5.5 mL of concentrated nitric acid was added to the white milk colloid obtained above, and the mixture was uniformly stirred and transferred. The reaction was carried out at 195 ° C for 24 hours in a 50 mL reactor equipped with a Teflon-lined reactor. After the reaction is completed, let it cool naturally to room temperature in the air, discard the upper liquid, and rinse the solid at the bottom of the kettle with hot water into the beaker, sonicate for 10 minutes, then let it stand for a few minutes. After the solid precipitates to the bottom of the beaker, discard the upper layer. The liquid was reheated and sonicated. After repeated operations for 3 times, the ethanol was ultrasonically dispersed in a beaker. Finally, the solid obtained by centrifugation was dried in an oven at 70 ° C for 10 hours, NaY. ₇₈ F _4: Yb. . ₂ , E _r . . . . ₂ Upconverting Nanoparticles (UCNPs) solid powder and storing for use;

[0032] The second step, amination: 20mg of UCNPs was dissolved in 60mL of isopropanol, sonicated for 40 minutes, then added 2.5mL of ammonia and 20mL of water, fully stirred at 35 ° C, then drop into a small bowl Dissolved in 50u L

TEOS (ethyl orthosilicate) in 20 mL of isopropanol, reacting 3 small hydrazines to form a suspension, and then adding 30 mL of isopropanol dissolved in 200 uL of APTES (3-aminopropyltriethoxysilane) dropwise In the suspension obtained above, one small reaction was carried out; after completion of the reaction, the mixture was aged for 2 hours at room temperature, and a precipitated solid was obtained by centrifugation, and the solid obtained by precipitation was washed three times with ethanol and at 60. After drying for 12 hours under C, the aminated upconverting nanomaterial is finally obtained;

[0033] The third step, avidinization: according to the glutaraldehyde method, the aminated nano-material obtained in the second step is connected with avidin to form an avidinized up-converting nanomaterial; [0034] In the fourth step, the avidinized up-converting nano material obtained in the third step is combined with the corresponding pathogen-specific aptamer to obtain an up-converting fluorescent probe;

[0035] The up-converted fluorescent probe obtained in the fourth step is dissolved in 5 ml of PBS (phosphate buffer) solution, and allowed to stand for 6 hours.

, kept at 4 ° C for use.

[0036] The preparation process of the magnetic probe is as follows:

[0037] In the first step, the magnetic beads are synthesized by a hydrothermal solvothermal method: using chlorinated high-iron hexahydrate as the iron source and 1,6-hexanediamine as the amino functionalizing agent, and weighing 1,6 hexamethylenediamine 6.5 g, anhydrous sodium acetate 2.0g, F eC13*6H20 l.Og dissolved in 30mL of ethylene glycol, added to a lOOmL round bottom flask, vigorously stirred at 50 ° C for about 0.5 吋 to the solution is more transparent, the solution is transferred to the band In a high-pressure reaction vessel lined with tetrafluoroethylene, it was placed at a high temperature of 198 ° C for 6 hours. After the reaction, it was taken out and naturally cooled to room temperature. The liquid in the reactor was poured into a beaker and washed alternately with absolute ethanol and pure water. Three times, the black solid at the bottom of the reactor was at 50. Drying overnight under C conditions to obtain aminated magnetic nanoparticles, that is, aminated magnetic beads;

[0038] The second step, avidinization: according to the glutaraldehyde method, the aminated magnetic beads obtained in the second step are connected with avidin to form avidinized magnetic beads;

[0039] In the third step, the avidinized magnetic beads obtained in the third step are combined with the corresponding pathogen-specific aptamers to obtain a magnetic probe;

[0040] The magnetic probe obtained in the third step was dissolved in 10 mL of O.Olmol/L PBS (phosphate buffer, pH 7.4) and stored at 4 ° C until use.

[0041] Example 1 Detection of Salmonella

[0042] After enrichment and cultivation of Salmonella, the medium is removed by centrifugation, and the concentration is determined by a plate method, and then the gradient is diluted into a suspension of different standard concentrations (cfu/ml); 5 μί of Salmonella aptamer is added, respectively. The lm L concentration is 1 mg/mL of avidinized magnetic beads and avidinized up-converting nanomaterials, shaken reaction for 12 hours, and finally added 2% BSA (bovine serum albumin) blocking solution to obtain magnetic properties. Probe and upconversion fluorescent probe; equal volume of gradient Salmonella standard concentration bacterial suspension was added to 200μ1 upconversion fluorescent probe and 80μ1 magnetic probe respectively, and after 40 minutes incubation at 37 ° C, a sandwich sandwich complex was formed. The combined up-converting fluorescent probe is magnetically separated, the supernatant is decanted, and the precipitate is resuspended in BB buffer, and the resuspended sandwich sandwich test solution is placed in cuvette 4 through infrared The light source excites the infrared light to contrast the color dish 4, and the fluorescence intensity of the sandwich sandwich test solution is measured, thereby drawing and being tested. The fluorescence intensity-concentration standard regression curve of Salmonella in liquid is shown in Fig. 3. In Fig. 3, the horizontal axis represents the concentration of the Salmonella suspension, and the number axis represents the fluorescence intensity of the sandwich sandwich test solution in the cuvette. The regression curve shows that the fluorescence intensity of the test solution in the cuvette has a linear relationship with the concentration of Salmonella, and the expression of the statistically fitted fluorescence intensity-concentration standard regression curve is: Y=169.66X-35.2, The square of the combining coefficient is 0.9964, and the detection limit is 5 cfu/ml; on the other hand, by the functional relationship of the statistical fit, the detection system of the present invention can be used to measure the concentration of Salmonella in an unknown concentration sample, and Peer and the classic flat method test results were tested and verified. Among them, the Salmonella aptamer adopts the Salmonella aptamer synthesized by Bioengineering Biotechnology (Shanghai) Co., Ltd.: 5'-biotin-C6-TAT GGC GGC GTC ACC CGA CGG GGA CTT ATG GAC ATT ACA G-3'.

[0043] The lake water, the river water, and the small groove water are respectively sampled, and the concentration of the Salmonella is detected by the conventional plate colony counting method and the detection system of the present invention, respectively, and the test results of the two are compared as follows:

Table 1 Comparison of the detection results of the plate colony counting method and the detection system of the present invention for Salmonella detection

[]

[0045] According to the above experimental data comparison, the detection system of the present invention is basically in agreement with the detection results of the classical plate colony counting method.

Example 2 Detection of Staphylococcus aureus

[0047] After the S. aureus is enriched and cultured, the medium is removed by centrifugation, and the concentration is determined by a plate method, and then the gradient is diluted into a suspension of different standard concentrations (cfu/ml); 5 μί golden yellow grapes are respectively taken. Cocci aptamer, add 1 mL of avidinized magnetic beads and avidinized upconversion nanomaterials with a concentration of 1 mg/mL, shake the reaction for 12 hours, and finally add 2% BSA (bovine serum albumin) to block. Liquid, get magnetic exploration Needle and upconversion fluorescent probes; equal volumes of gradient Staphylococcus aureus standard concentration bacterial suspension were added to 2 ΟΟμΙ upconversion fluorescent probes and ΙΟΟμΙ magnetic probes, respectively. After incubating for 40 minutes at C, a sandwich sandwich complex was formed, magnetic separation was performed with an unbound up-conversion fluorescent probe, the supernatant was discarded, and the sandwich was resuspended with BB buffer after precipitation. The sandwich composite test solution is placed in the cuvette 4, and the infrared light source is used to illuminate the infrared light contrasting plate 4, and the sandwich sandwich composite test solution is subjected to fluorescence intensity measurement to thereby determine the fluorescence intensity-concentration standard of the Staphylococcus aureus. Regression curve, the fluorescence intensity-concentration standard regression curve of Staphylococcus aureus is shown in Figure 4. In Figure 4, the horizontal axis represents the concentration of the Staphylococcus aureus suspension, and the number axis represents the sandwich sandwich compound in the cuvette. The fluorescence intensity of the test solution is detected. From the regression curve, the fluorescence intensity of the test solution in the cuvette is linearly related to the concentration of Staphylococcus aureus in the test liquid, and the fluorescence intensity-concentration is statistically fitted. standard regression curve expression is: Y = 118.5X + 15.6, squares fitting coefficient of 0.9936, the detection limit was 8cfu / _ml; on the other hand, through The statistical fit function formula, i.e., the detection system can be used according to the present invention against Staphylococcus aureus concentration of the unknown concentration in the sample was measured and verified with the detection result of the detection classical inch plate method .. Among them, S. aureus aptamer adopts S. aureus aptamer synthesized by Bioengineering Biotechnology (Shanghai) Co., Ltd.: 5'-biotin-C6-GCA ATG

[0048] The lake water, the river water, and the small groove water are separately sampled, and the concentration of the Staphylococcus aureus is detected by the conventional plate colony counting method and the detection system of the present invention, respectively, and the test results are compared as shown in Table 2 below:

[0049] Table 1 comparison between the plate counting method and the detection system of the present invention for the detection results of Staphylococcus aureus

[]

Sample plate counting method portable up-conversion fluorescence detection

^s-s . - 3⁄4■ B _L_ a 2ii soil 'ό , 2 river water 75 soil 2. 8

4, inquiry water 239 ± . 3 226 soil 3. 2 According to the above experimental data comparison, the detection system of the present invention is basically consistent with the detection results of the classical plate colony counting method.

Claims

Claim

The fluorescent biological detection system comprises a light emitting module (1), a fluorescent receiving module (2) and a data processing module (3), characterized in that: a cuvette is arranged between the light emitting module (1) and the fluorescent receiving module (2) (4) a detection test solution containing an up-converting fluorescent material mark in the cuvette (4); the light-emitting module (1) includes an infrared excitation light source, and the excitation light beam emitted from the infrared excitation light source is irradiated through the excitation light path On the cuvette (4) containing the test solution, the fluorescence emitted by the test solution is received by the receiving light route fluorescent receiving module (2), and the excitation light path is arranged at a straight line or a right angle with the receiving optical path; The fluorescence receiving module (2) is a light intensity sensor, and the output end of the light intensity sensor is connected to the data processing module (3).

The fluorescent biological detection system according to claim 1, wherein: said detection test solution comprises a combination of an up-converting fluorescent probe and a pathogen-specific aptamer, a magnetic probe and a pathogen-specific aptamer, The composition of the bacteria to be tested.

The fluorescent biodetection system according to claim 2, wherein: said up-converting fluorescent probe is obtained by subjecting up-converting fluorescent material to amination and avidinization, and said up-converting fluorescent material is NaY0.78F4: Yb0 .2, Er0.02 nanoparticles.

The fluorescent biological detection system according to claim 2, wherein: said magnetic probe is an avidinized magnetic bead, said magnetic bead being ferrous chloride hexahydrate as an iron source, 1, 6 - hexane The amine is synthesized as an amino functionalizing agent by a hydrothermal-solvent method.

The fluorescent biological detection system according to claim 1, wherein: the data processing module (3) comprises an amplifying circuit module, an anti-interference module, a single chip signal processing module, and a liquid crystal display module, wherein the amplifying circuit module is used for amplifying Receiving a fluorescent signal, the anti-interference module is configured to convert the voltage signal output by the light intensity sensor into a current signal transmission, and convert it into a voltage signal at the line terminal, and the single-chip signal processing module performs data operation processing and passes the result The liquid crystal display module performs an actual display.

The fluorescent biological detection system according to claim 5, wherein: said data processing module (3) comprises a communication module, and said communication module is used for remote communication between the single chip microcomputer and the upper computer. [Claim 7] The fluorescent biological detection system according to claim 1, wherein: the infrared excitation light source uses 980 nm near-infrared light.