CN110464361B - Method for potential fingerprint display by enzymatic reaction system - Google Patents
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- A61B5/117—Identification of persons
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- A61B5/1172—Identification of persons based on the shapes or appearances of their bodies or parts thereof using fingerprinting
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Abstract
The invention relates to the technical field of trace detection, and particularly discloses a method for displaying a potential fingerprint by using an enzymatic reaction system. The invention has the advantages that the invention is sensitive and rapid, the composition of the chromogenic reagent is not complex, no toxicity is generated to operators, the invention can also realize 'hidden reconnaissance', and the invention is expected to be applied in the aspects of criminal reconnaissance and the like.
Description
Technical Field
The invention relates to the technical field of trace detection, in particular to a method for using an enzymatic reaction system for potential fingerprint display.
Background
The fingerprints are different in people and are not changed all the time, and the contact object is marked. The method has the advantages that the method scientifically and correctly discovers, extracts, shows and identifies the fingerprints, has important practical effects on carrying out investigation work and punishing crimes, and is an important basis for case investigation by extracting and identifying the fingerprints left on the surface of a guest.
Current methods of identifying potential fingerprints can be divided into three major categories: the development and improvement of the current technology mainly focus on adopting novel characterization means to increase the sensitivity of the detection and different application ranges, and in Chinese patent (CN201410146802.3), Wanglihua and the like adopt various fluorescence quenching systems to perform potential fingerprint visualization. In chinese patent (CN201810189499.3), the inventor prepared a magnetic fluorescent powder for latent fingerprint visualization to improve the sensitivity, but the preparation process is complicated. Chinese patent (CN201710888373.0) prepared a special material: the black body radiation luminescent material can reduce background fluorescence to obtain clearer fingerprint images, but the preparation process of the material is complicated.
The enzyme is used as a biocatalyst, and has high catalytic reaction efficiency, strong specificity and high sensitivity. The Chinese patent (CN201710045379.1) utilizes enzyme to display latent fingerprints, but the latent fingerprints need to be transferred twice, and then are marked by antibodies and then are displayed by related reagents, and the whole operation process is very complex and takes a long time.
At present, no report is found for showing latent fingerprints by catalyzing the reaction of trace substances in the latent fingerprints in a short time at normal temperature and normal pressure.
Disclosure of Invention
In view of the deficiencies of the prior art, it is an object of the present invention to provide an enzymatic reaction system for latent fingerprint visualization. Sweat is the main component of latent fingerprints, most of moisture in the sweat contains substances such as lactic acid, amino acid, protein, sodium chloride and the like, and the content of the lactic acid in the sweat is about 6.6 multiplied by 10 according to literature data-3mol/L. The lactate dehydrogenase can catalyze the reaction of lactic acid to generate pyruvic acid, and the detection limit of lactic acid in the reaction is 2.25X 10-5mol/L; due to the specificity of the enzymatic reaction, other coexisting components in the latent fingerprint do not interfere with the reaction, so that the lactic acid in the latent fingerprint can be directly measured by using the method. The latent fingerprint display method with low cost, high speed, high sensitivity and simple operation can be established by utilizing the enzymatic reaction. The method can rapidly display the latent fingerprints, the composition of the reagent is not complex, the display operation is simple, and no toxicity is generated to operators.
In order to achieve the above technical object, an enzymatic reaction system of the present invention is used in a method for latent fingerprint development, comprising the steps of:
(1) preparing an enzymatic reaction system color developing solution, and placing the enzymatic reaction system color developing solution in a refrigerator for later use;
(2) spraying or coating the enzymatic reaction system color developing liquid on a fingerprint object which has a fingerprint but does not contain fluorescent substances;
(3) and developing under ultraviolet irradiation.
The enzymatic reaction system color developing solution comprises: 50-80U/mL lactate dehydrogenase, 0.007-0.009mol/L oxidized nicotinamide adenine dinucleotide and Tris-HCl-hydrazine hydrate buffer solution.
The preparation method of the Tris-HCl-hydrazine hydrate buffer solution comprises the following steps: configuring a buffer solution of 0.05mol/L of LTris-HCl at pH9.0 according to an appendix of a conventional biochemical experimental book, and adding hydrazine hydrate to the buffer solution to make the final concentration of the hydrazine hydrate be 1.2-1.8 mol/L (preferably 1.5 mol/L);
the hydrazine hydrate is added in the form of hydrazine hydrate solution with hydrazine hydrate concentration of above 50% (mass percentage).
The order of spraying or coating the chromogenic solution of the enzymatic reaction system on the fingerprint object is as follows: spraying or coating Tris-HCl-hydrazine hydrate buffer solution and oxidized nicotinamide adenine dinucleotide, and spraying or coating lactic dehydrogenase, wherein the sequence of the Tris-HCl-hydrazine hydrate buffer solution and the oxidized nicotinamide adenine dinucleotide is not limited, and the Tris-HCl-hydrazine hydrate buffer solution and the oxidized nicotinamide adenine dinucleotide can be sprayed or coated simultaneously.
Preferably, a proper amount of water-soluble compound with poor fluidity, such as glycerol and polyethylene glycol 400(PEG400), can be added into the color developing solution of the enzymatic reaction system to increase the viscosity of the system, so that the developing result can be clearer, and the water-soluble compound is added into a Tris-HCl-hydrazine hydrate buffer solution instead of part of water.
Preferably, the fingerprint is a latent sweat fingerprint.
Preferably, the fingerprint object is writing paper, kraft paper, filter paper, coated paper, newspaper and the like without fluorescent components.
The reaction mechanism of the enzymatic reaction system of the present invention is as follows:
lactic acid in oxidized form nicotinoylAmine adenine dinucleotide (NAD)+) Pyruvate and reduced Nicotinamide Adenine Dinucleotide (NADH) are produced in the presence of Lactate Dehydrogenase (LDH) as a catalyst. Adding Tris-HCl-hydrazine hydrate buffer solution into the reaction system can enable the pyruvic acid generated by the reaction to be converted continuously, and enable the lactic acid in the detection sample to react completely. NADH produced by the reaction is a strong fluorescent substance, and substrate NAD+Since there is no fluorescence, the presence or absence of lactate can be determined from the presence or absence of a fluorescence signal, and the content of lactate in the sample can be calculated by measuring the rate of change in the NADH fluorescence value.
Compared with the prior art, the method has the advantages and beneficial effects as follows:
1. the latent fingerprints can be quickly displayed; the display operation is simple.
2. The composition of the developing reagent is not complex, and the cost is low; it has no toxicity to operator.
3. Colored substances can not be formed in the reaction, the latent sweat fingerprints can be observed only under the light irradiation of special wavelengths, and the abnormality on the articles can not be seen under natural light or common lamplight, so that the hidden investigation can be realized, and the latent fingerprint can be expected to be applied to the displaying aspect of latent fingerprints such as criminal investigation.
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FIG. 1 is a fingerprint image of example 1 appearing on writing paper, wherein FIG. 1(a) shows that background light is strong to the naked eye without a filter after being excited by ultraviolet light with a wavelength of 365 nm; FIG. 1(b) shows that the interference of background light is weak when the filter is present after the excitation of 365nm ultraviolet light.
FIG. 2 is a diagram of fingerprints appearing on writing paper in example 2.
FIG. 3 is a diagram of fingerprints appearing on writing paper in example 3.
Figure 4 is a fingerprint image of example 4 appearing on kraft paper.
FIG. 5 is a graph of fingerprints appearing on the filter paper in example 5.
Detailed Description
The technical scheme provided by the invention is explained in detail by combining the specific embodiment as follows:
NAD+is beta-smokeAmide adenine dinucleotide, available from bioscience, beijing solebao, BR, 1g, as powder. CAS number: 53-84-9.
Lactate dehydrogenase (rabbit muscle), BR, 30U/mg, powder, available from shanghai-derived leaf biotechnology limited. CAS number: 9001-60-9.
Example 1
A method for using an enzymatic reaction system for latent fingerprint visualization comprises the following steps:
1. preparing a natural fingerprint sample: please take the testers who provide the fingerprints to clean their handles, dry them in the air, wear plastic gloves, cover them for 10 minutes, press the hands of the testers with fingers on the writing paper without fluorescent components, and leave them for 4 hours for use (simulating the actual situation of criminal investigation.
2. Preparing a color developing solution:
1) Tris-HCl-hydrazine hydrate buffer (hydrazine hydrate concentration 1.5 mol/L): adding 5mL0.1mol/LHCl solution into 25mL0.2mol/LTris solution, adding water to 100mL to obtain Tris-HCl buffer solution with pH of 9.0 and 0.05mol/L, and adding 9.38mL of 85% (mass percentage, the same below) hydrazine hydrate solution into the buffer solution to obtain Tris-HCl-hydrazine hydrate buffer solution with the hydrazine hydrate concentration of 1.5 mol/L.
2)0.007mol/LNAD+: 0.046g of NAD+Adding into 10mL distilled water.
3)50U/mL lactate dehydrogenase: dissolving 10mg of lactate dehydrogenase in 6mL of distilled water to obtain the final product.
After the solutions are prepared, the mixed solution is placed in a refrigerator for standby.
3. And (3) showing: sequentially dipping Tris-HCl-hydrazine hydrate buffer solution and 0.007mol/LNAD by using a fine hairbrush+And (3) smearing the fingerprint part of the sample prepared in the step (1) with 50U/mL lactate dehydrogenase in the same direction, smearing the fingerprint part in a small amount without repeated smearing, slightly drying the fingerprint part, irradiating and exciting the fingerprint part by using an ultraviolet lamp with the wavelength of 365nm, and displaying the fingerprint, and taking a picture and recording the fingerprint. In order to eliminate background light interference, the fingerprint searching can be assisted by a 450nm filter.
As can be seen from fig. 1, a part of fingerprint lines can be displayed on the writing paper, wherein (a) after being excited by ultraviolet light with a wavelength of 365nm, when no optical filter is arranged, the background light is strong when the naked eye sees; (b) after being excited by ultraviolet light with the wavelength of 365nm, the interference of background light is weak when the light filter is arranged.
Example 2
A glycerol-added enzymatic reaction system is used for the method of latent fingerprint development, and the steps are the same as those of example 1, except that the buffer is prepared differently.
This example uses a Tris-HCl-glycerol-hydrazine hydrate buffer (with a hydrazine hydrate concentration of 1.5 mol/L): adding 5mL0.1mol/LHCl solution into 25mL0.2mol/LTris solution, adding 20mL of glycerol, adding water to 100mL to obtain pH9.0, 0.05 mol/LTris-HCl-glycerol buffer solution, and adding 9.38mL of 85% hydrazine hydrate solution into the buffer solution to obtain Tris-HCl-glycerol-hydrazine hydrate buffer solution with the hydrazine hydrate concentration of 1.5 mol/L.
As can be seen from fig. 2, a clear fingerprint pattern can be shown on the writing paper.
Example 3
An enzymatic reaction system with PEG400 was used for latent fingerprint development, which was performed in the same manner as in example 1, except that the buffer was formulated differently.
This example uses a Tris-HCl-PEG-hydrazine hydrate buffer (with a hydrazine hydrate concentration of 1.5 mol/L): adding 5mL0.1mol/LHCl solution into 25mL0.2mol/LTris solution, adding 15mLPEG400, adding water to 100mL to obtain pH9.0, 0.05mol/LTris-HCl-PEG buffer solution, and adding 9.38mL 85% hydrazine hydrate solution into the buffer solution to obtain Tris-HCl-PEG-hydrazine hydrate buffer solution with the hydrazine hydrate concentration of 1.5 mol/L.
As can be seen from fig. 3, a clear fingerprint pattern can be shown on the writing paper.
Example 4
A method for using an enzymatic reaction system for latent fingerprint visualization comprises the following steps:
1. preparing a natural fingerprint sample: please refer to the examiner who provides the fingerprint to clean his/her hands, dry his/her hands by air, wear plastic gloves, cover his/her sweat for 10 minutes, press the gloves with fingers on the kraft paper without fluorescent component, and leave for 4 hours for use (simulating the actual situation of criminal investigation in case).
2. Preparing a color developing solution:
1) Tris-HCl-glycerol-hydrazine hydrate buffer (hydrazine hydrate concentration 1.2 mol/L): adding 5mL0.1mol/LHCl solution into 25mL0.2mol/LTris solution, adding 20mL of glycerol, adding water to 100mL to obtain Tris-HCl-glycerol buffer solution with pH of 9.0 and 0.05mol/L, and adding 7.37mL of 85% hydrazine hydrate solution into the buffer solution to obtain Tris-HCl-glycerol-hydrazine hydrate buffer solution with the hydrazine hydrate concentration of 1.2 mol/L.
2)0.009mol/LNAD+: 0.060g NAD+Adding into 10mL distilled water.
3)80U/mL lactate dehydrogenase: dissolving 16mg of lactate dehydrogenase in 6mL of distilled water to obtain the lactate dehydrogenase.
After the solutions are prepared, the mixed solution is placed in a refrigerator for standby.
3. And (3) showing: dipping Tris-HCl-glycerol-hydrazine hydrate buffer solution and 0.009mol/LNAD in sequence by using a fine hair brush+And (3) smearing the fingerprint part of the sample prepared in the step (1) by 80U/mL lactate dehydrogenase in the same direction, smearing the fingerprint part in a small amount without repeated smearing, slightly drying the fingerprint part, irradiating and exciting the fingerprint part by using an ultraviolet lamp with the wavelength of 365nm to show the fingerprint, and taking a picture for recording.
As can be seen from fig. 4, a relatively clear fingerprint pattern can be shown on the kraft paper.
Example 5
A method for using an enzymatic reaction system for latent fingerprint visualization comprises the following steps:
1. preparing a natural fingerprint sample: please take the testers who provide fingerprints to clean their hands, dry them in the air, wear plastic gloves, cover them for 10 minutes, press them on the filter paper without fluorescence component, and leave them for 4 hours (simulating the actual situation of criminal investigation.
2. Preparing a color developing solution:
1) Tris-HCl-PEG-hydrazine hydrate buffer (hydrazine hydrate concentration 1.8 mol/L): adding 5mL0.1mol/LHCl solution into 25mL0.2mol/LTris solution, adding 15mLPEG400, adding water to 100mL to obtain Tris-HCl-PEG buffer solution with pH of 9.0 and 0.05mol/L, and adding 11.47mL 85% hydrazine hydrate solution into the buffer solution to obtain Tris-HCl-PEG-hydrazine hydrate buffer solution with hydrazine hydrate concentration of 1.8 mol/L.
2)0.009mol/LNAD+: 0.060g NAD+Adding into 10mL distilled water.
3)50U/mL lactate dehydrogenase: dissolving 10mg of lactate dehydrogenase in 6mL of distilled water to obtain the final product.
After the solutions are prepared, the mixed solution is placed in a refrigerator for standby.
3. And (3) showing: dipping Tris-HCl-PEG-hydrazine hydrate buffer solution and 0.009mol/LNAD in sequence by using a fine hair brush+And (3) smearing the fingerprint part of the sample prepared in the step (1) with 50U/mL lactate dehydrogenase in the same direction, smearing the fingerprint part in a small amount without repeated smearing, slightly drying the fingerprint part, irradiating and exciting the fingerprint part by using an ultraviolet lamp with the wavelength of 365nm, and displaying the fingerprint, and taking a picture and recording the fingerprint.
As can be seen from fig. 5, a relatively clear fingerprint pattern can be displayed on the filter paper.
Claims (5)
1. A method for an enzymatic reaction system for latent fingerprint visualization, characterized by the steps of:
(1) preparing an enzymatic reaction system color developing solution, and placing the enzymatic reaction system color developing solution in a refrigerator for later use;
(2) spraying or coating the enzymatic reaction system color developing liquid on a fingerprint object which has a fingerprint but does not contain fluorescent substances;
(3) developing under ultraviolet irradiation;
the enzymatic reaction system color developing solution comprises: 50-80U/mL lactate dehydrogenase, 0.007-0.009mol/L oxidized nicotinamide adenine dinucleotide and Tris-HCl-hydrazine hydrate buffer solution;
the order of spraying or coating the chromogenic solution of the enzymatic reaction system on an object is as follows: spraying or coating a Tris-HCl-hydrazine hydrate buffer solution and oxidized nicotinamide adenine dinucleotide, and finally spraying or coating lactic dehydrogenase, wherein the sequence of the Tris-HCl-hydrazine hydrate buffer solution and the oxidized nicotinamide adenine dinucleotide is not limited, or spraying or coating simultaneously;
the preparation method of the Tris-HCl-hydrazine hydrate buffer solution comprises the following steps: firstly, preparing a Tris-HCl buffer solution with the pH value of 9.0 and the concentration of 0.05mol/L, and then adding hydrazine hydrate into the Tris-HCl buffer solution to ensure that the final concentration of the hydrazine hydrate is 1.2-1.8 mol/L;
the Tris-HCl buffer solution with the pH value of 9.0 and the concentration of 0.05mol/L contains glycerol and/or polyethylene glycol 400 accounting for 40 percent of the volume of the Tris-HCl buffer solution with the pH value of 9.0 and the concentration of 0.05 mol/L.
2. The method of claim 1, wherein the Tris-HCl buffer at ph9.0, 0.05mol/L contains glycerol and/or polyethylene glycol 400 in an amount of up to 25% by volume of the Tris-HCl buffer at ph9.0, 0.05 mol/L.
3. The method according to any one of claims 1-2, wherein: the oxidized nicotinamide adenine dinucleotide is beta-nicotinamide adenine dinucleotide.
4. The method according to any one of claims 1-2, wherein: the fingerprint is a latent sweat fingerprint.
5. The method according to any one of claims 1-2, wherein: the fingerprint object is one of writing paper, kraft paper, filter paper, coated paper or newspaper without fluorescent components.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103919558A (en) * | 2014-04-13 | 2014-07-16 | 中南民族大学 | Method of adopting fluorescence quenching system to potential fingerprint display |
| CN104783806A (en) * | 2015-05-05 | 2015-07-22 | 哈尔滨师范大学 | Method for displaying latent fingerprints through fluorescent copper sol |
| CN106645062A (en) * | 2016-11-30 | 2017-05-10 | 青岛农业大学 | Method for laten fingerprint fluorescence development by applying aptamer and lysozyme for specific recognition and formation of G-quadruplex |
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| US10552659B2 (en) * | 2016-08-04 | 2020-02-04 | Howard A. Harris | Latent fingerprint development on porous surfaces |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103919558A (en) * | 2014-04-13 | 2014-07-16 | 中南民族大学 | Method of adopting fluorescence quenching system to potential fingerprint display |
| CN104783806A (en) * | 2015-05-05 | 2015-07-22 | 哈尔滨师范大学 | Method for displaying latent fingerprints through fluorescent copper sol |
| CN106645062A (en) * | 2016-11-30 | 2017-05-10 | 青岛农业大学 | Method for laten fingerprint fluorescence development by applying aptamer and lysozyme for specific recognition and formation of G-quadruplex |
Non-Patent Citations (1)
| Title |
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| 乳酸含量的酶法测定;庹浔;《中国优秀博硕士学位论文全文数据库（硕士）工程科技Ⅰ辑》;20080315;第18-24页 * |
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