Colloidal gold test paper for rapidly detecting trace element manganese in food

Abstract

The utility model discloses colloidal gold test paper for rapidly detecting trace element manganese in food, which comprises a support gasket 1, a nitrocellulose membrane 2 positioned on the support gasket 1, a Mn antibody detection line 3 coated in the middle of the nitrocellulose membrane 2 and control lines 4 respectively arranged on two sides of the detection line 3, and a water absorption pad 5 is connected above one end of the nitrocellulose membrane 2. A conjugate gasket 6 is connected above the other end of the sample pad 7 and is connected with a sample pad 7; the detection line 3 is coated with a monoclonal mouse anti-Mn antibody, the conjugate gasket 6 is coated with gold-labeled BSA-Mn and a control line conjugate, the control line 4 is coated with an anti-DNP antibody, and the control line conjugate is DNP-BSA labeled colloidal gold.

Description

Technical Field

[0001] The utility model belongs to the field of food industry analysis, and specifically relates to a colloidal gold test paper used for detecting trace element manganese in food. Background Art

[0002] Manganese, with the symbol Mn, is an essential trace element for the human body. It is found in the cerebral cortex, kidney, pancreas, and mammary gland. Mn can promote the growth and development of human bones and protect the integrity of mitochondria in cells. It participates in the synthesis and activation of enzymes, participates in the metabolism of sugar and fat in the human body, catalyzes sebum metabolism, promotes the excretion of harmful substances in the skin, and prevents dry skin. It accelerates the synthesis of protein, vitamin B, and C, catalyzes hematopoiesis, and regulates endocrine. It promotes the accumulation of vitamin B6 in the liver, thereby enhancing the body's ability to resist inflammation. Manganese deficiency can cause many adverse effects. For example, manganese deficiency in children can lead to growth stagnation and bone deformities; manganese deficiency in adults can lead to loss of appetite, decreased reproductive function, and even central nervous system symptoms.

[0003] Mn has a wide range of food sources in nature, such as meat (pork, mutton, beef, chicken, fish, etc.), eggs, milk and dairy products, vegetables, fruits, aquatic products, edible fungi, seasonings (pepper, chili, vinegar), liquor (distilled liquor), tea, Chinese medicinal materials, tonics (Chinese oral liquid, etc.), etc. Therefore, the qualitative analysis of Mn is of great significance in clinical and food science.

[0004] The existing methods for detecting Mn are as follows: the first method given in GB 5009.242-2017 "National Food Safety Standard for the Determination of Manganese in Food" is flame atomic absorption spectrometry, and the second method given in GB 5009.268-2016 "National Food Safety Standard for the Determination of Multiple Elements in Food" is inductively coupled plasma emission spectrometry and the third method is inductively coupled plasma mass spectrometry. These three methods are complicated to operate, have long detection cycles, and poor repeatability, and the success rate of inexperienced laboratories is low. In addition, flame atomic absorption spectrometry (atomic absorption spectrometer), inductively coupled plasma emission spectrometry (inductively coupled plasma emission spectrometer) and inductively coupled plasma mass spectrometry (inductively coupled plasma mass spectrometer) are expensive instruments, have high operating costs, and take long analysis times, and cannot meet the needs of simple operation and rapid detection.

[0005] Immune colloidal gold technique is a new type of immunolabeling technique that uses colloidal gold as a tracer marker for antigen-antibody. Colloidal gold is polymerized into gold particles of a specific size by chloroauric acid (HAuCl4) under the action of reducing agents such as white phosphorus, ascorbic acid, sodium citrate, trisodium citrate, tannic acid, etc., and becomes a stable colloidal state due to electrostatic action, called colloidal gold. Colloidal gold carries a negative charge in a weak alkaline environment and can form a strong bond with the positively charged groups of protein molecules. Since this bond is an electrostatic bond, it does not affect the biological properties of the protein. In addition to binding to proteins, colloidal gold can also bind to many other biological macromolecules. Based on some physical properties of colloidal gold, such as high electron density, particle size, shape and color reaction, plus the immune and biological properties of the conjugate, colloidal gold is widely used in immunology, histology, pathology and cell biology.

[0006] Visual detection based on colloidal gold provides a new technical platform for the qualitative analysis of Mn. Colloidal gold particles have the characteristics of high electron density. When the macromolecules bound to the colloidal gold particles are aggregated in large quantities at the corresponding ligands, red or pink spots visible to the naked eye will be formed. Colloidal gold test strips fix various reaction reagents on the test strips in the form of strips. The sample solution moves from one end of the test strip to the other end through capillary action, and the analyte in the sample reacts specifically with the receptors on the chromatographic material for the analyte. The sample bound to colloidal gold is intercepted and aggregated in a certain area of ​​the chromatographic material, and the color can be intuitively obtained by visual inspection. This study uses the principle of colloidal gold to develop colloidal gold for rapid detection of Mn by combining colloidal gold particles coupled with BSA or OVA with monoclonal mouse anti-Mn antibodies fixed on nitrocellulose (NC) filter membranes.

[0007] In view of the fact that there is no rapid Mn detection method at present, the purpose of the utility model is to provide a colloidal gold test paper that can be used for rapid detection of Mn and a colloidal gold test paper box that can be used for rapid detection of Mn. The colloidal gold test paper and the colloidal gold test paper box have the advantages of convenience, simplicity of operation, and accurate results. Utility Model Content

[0008] The utility model provides a colloidal gold test paper for rapidly detecting trace element manganese in food, which has the advantages of convenience, speed, simple operation, accurate results, etc.

[0009] The technical solution of the utility model is:

[0010] A colloidal gold test paper for rapid detection of trace element manganese in food, comprising a supporting gasket 1, a nitrocellulose membrane 2 located on the supporting gasket 1, a Mn antibody detection line 3 coated in the middle of the nitrocellulose membrane 2, and control lines 4 respectively arranged on both sides of the Mn antibody detection line 3, wherein an absorbent pad 5 is connected to the top of one end of the nitrocellulose membrane 2, and a conjugate pad 6 is connected to the top of the other end, and the conjugate pad 6 is connected to a sample pad 7; the characteristic is that the Mn antibody detection line 3 is coated with a monoclonal mouse anti-Mn antibody, and the conjugate pad 6 is coated with gold-labeled BSA-Mn and a control line conjugate.

[0011] Furthermore, the control line 4 is coated with anti-DNP antibodies.

[0012] Furthermore, a fixing pad 8 is provided above the sample pad 7, and the fixing pad can be a single-sided adhesive tape or a double-sided adhesive tape.

[0013] Furthermore, the control line conjugate is colloidal gold labeled with DNP-BSA.

[0014] The colloidal gold test paper for rapidly detecting trace element manganese in food of the utility model also includes a shell 9 wrapping the colloidal gold test paper, and windows for exposing a sample pad 7, a Mn antibody detection line 3 and two control lines 4 are respectively provided on the front of the shell.

[0015] The colloidal gold test paper for quickly detecting the trace element manganese in food of the utility model has the advantages of being convenient to carry and store.

[0016] The colloidal gold test paper for rapid detection of trace element manganese in food described in the utility model adopts the principle of two-way lateral flow immune reaction and is prepared by double antibody sandwich method. During the test, the Mn in the sample and the BSA-Mn colloidal gold conjugate on the coupling pad are chromatographed and flowed on the nitrocellulose membrane at the same time. When the Mn in the sample and the BSA-Mn colloidal gold conjugate are chromatographed to the detection area (Mn detection line) on the nitrocellulose membrane, they react with the Mn antibody pre-coated on the nitrocellulose membrane and are thus fixed on the Mn antibody detection line of the nitrocellulose membrane. The more complexes there are on the Mn antibody detection line, the higher the optical density value on the strip. At the same time, during the detection process, the control line conjugate will also be chromatographed on the nitrocellulose membrane with the sample. When chromatographed to the control line of the nitrocellulose membrane, the DNP-BSA colloidal gold will react with the anti-DNP antibody pre-coated on the nitrocellulose membrane and be fixed on the control line. Since the colloidal gold is red, the DNP-BSA colloidal gold immune complex in the anti-DNP fixed on the control line of the nitrocellulose membrane will show red. When the sample does not contain Mn, only two control lines are displayed. When the sample contains Mn, the Mn antibody detection line will be clearly displayed. Further, after the reaction is completed, a colloidal gold card reader (Guangdong Dayuan Oasis Food Safety Technology Co., Ltd.) can be used to perform concentration detection and display the results. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1 The utility model is a structural schematic diagram of a colloidal gold test paper for rapidly detecting trace element manganese in food.

[0018] Among them, 1: support pad, 2: nitrocellulose membrane, 3: Mn antibody detection line, 4: control line, 5: water absorption pad, 6: conjugate pad, 7: sample pad, 8: fixed pad.

[0019] Figure 2 It is a structural schematic diagram of another implementation mode of the colloidal gold test paper for rapidly detecting the trace element manganese in food described in the utility model.

[0020] Among them, 7: sample pad, 3: Mn antibody detection line, 4: control line, 9: shell. DETAILED DESCRIPTION

[0021] The colloidal gold test paper for rapid detection of Mn described in the utility model is described in detail below in conjunction with the accompanying drawings, which should not be construed as limiting the utility model. The materials and reagents used in the following embodiments are all commercially available unless otherwise specified. The utility model is applicable to the qualitative detection of Mn in: meat (pork, mutton, beef, chicken, fish, etc.), eggs, milk and dairy products, vegetables, fruits, aquatic products, edible fungi, seasonings (peppercorns, chili peppers, vinegar), liquor (distilled liquor), tea, Chinese medicinal materials, tonics (Chinese oral liquid, etc.), etc.

[0022] [Example 1] Preparation of colloidal gold test paper for rapid detection of Mn

[0023] Preparation Figure 1 The colloidal gold test paper structure shown includes a support pad 1, a nitrocellulose membrane 2 located on the support pad 1, a Mn antibody detection line 3 coated in the middle of the nitrocellulose membrane 2, and control lines 4 respectively arranged on both sides of the Mn antibody detection line 3, wherein the nitrocellulose membrane 2 is connected to a water absorbent pad 5 above one end and a conjugate pad 6 above the other end, and the conjugate pad 6 is connected to a sample pad 7; characterized in that the Mn antibody detection line 3 is coated with monoclonal mouse anti-Mn antibodies, and the conjugate pad 6 is coated with gold-labeled BSA-Mn and control line conjugates. The control line 4 is coated with anti-DNP antibodies, and the control line conjugate is colloidal gold labeled with DNP-BSA.

[0024] The nitrocellulose membrane 2 is used to fix the coated antibody in the colloidal gold test paper and is also the place where the immune reaction occurs; the Mn antibody detection line 3 is obtained by diluting the monoclonal mouse anti-Mn antibody to a concentration of 0.2-0.5 mg / mL, preferably 0.2 mg / mL, using a coating buffer [(carbonate buffer CBS, pH 9.6 0.1 mol / L): weigh 0.75 g Na2CO3, 1.466 g NaHCO3 and dilute to 500 mL with distilled water.], taking 1 uL / cm and drawing a line on the nitrocellulose membrane 2, and drying to obtain; the control line 4 is obtained by diluting the anti-DNP antibody to a concentration of 0.2-0.5 mg / mL, preferably 0.2 mg / mL, using a 0.01 mol / L PBS (pH 7.4) buffer containing 3% methanol and 0.1% Tween-20, taking 1 uL / cm and drawing a line on the nitrocellulose membrane 2, and drying to obtain.

[0025] The raw material of the coupling gasket 6 is a glass fiber filter membrane. The glass fiber filter membrane used to prepare the coupling gasket is placed in a pre-blocking buffer (containing 0.1% BSA, 5% sucrose, 0.5% PEG20000, 0.1% Tween-20 in 0.01mol / L PBS (pH7.4) buffer) and soaked for 5 minutes, then taken out, dried, and gold-labeled BSA-Mn and control line couplings are coated on the coupling gasket 6 using a Biodot coating instrument. The coating amount is 1uL / mm, and dried. Preferably, the pre-blocking buffer of the coupling gasket 6 contains 0.8% casein, 4% bovine serum albumin, 0.007% boric acid, and 0.1% PEG.

[0026] The sample pad 7 can play a preliminary filtering role for the liquid sample. Soak the sample pad in 141 blocking solution (BSA with a mass fraction of 0.2%-0.4%, 0.01mol / L PBS (pH7.4) buffer) for 5 minutes, and dry it at 37°C for 4.5 hours. Preferably, the 141 blocking solution contains: BSA with a mass fraction of 0.4%, 0.01mol / L PBS (pH7.4) buffer, and the above components are mixed according to the following steps: Figure 1 The structure shown is pasted on the support pad 1 to obtain a colloidal gold test paper for rapid detection of Mn.

[0027] [Example 2] Preparation of test samples:

[0028] 1. Fresh meat

[0029] Take 250g of the edible part of meat (pork, mutton, beef, chicken, fish, etc.), homogenize the sample with a homogenizer, and filter the homogenized sample with 4 layers of filter paper. Take 1.0g and add 5mL of mixed acid, cover and place for 1h or overnight, and place it on an adjustable temperature-controlled electric heating furnace for digestion. When white smoke appears and the digestion liquid is colorless and transparent or slightly yellow, cool it, dilute it to 25mL with 0.01mol / LPBS (pH7.4) solution, mix it for later use, take 100μL and test it on the prepared test paper, and directly observe the C and T line after 10min. The detection limit for fresh meat is 1μg / kg.

[0030] 2. Eggs

[0031] Mix the egg yolk and egg white evenly, filter with 4 layers of filter paper, take 1.0g and add 5mL of mixed acid, cover and place for 1h or overnight, place on an adjustable temperature-controlled electric heating furnace for digestion, wait until white smoke appears and the digestion liquid is colorless and transparent or slightly yellow, cool, dilute to 25mL with 0.01mol / L PBS (pH7.4) solution, mix well for later use, take 100μL and test on the prepared test paper, and directly observe the C and T lines after 10min. The detection limit for fresh eggs is 1μg / kg.

[0032] 3. Dairy products

[0033] Dairy product sampling method:

[0034] Raw milk / finished milk / yogurt: Measure 10 mL of raw milk and place it in a 100 mL conical flask.

[0035] Milk powder: Use an electronic balance to accurately weigh 1.0g, place in a 100mL conical flask, add 9mL of purified water, shake and mix, and set aside

[0036] Cheese: Grind the cheese, weigh 1.0g accurately with an electronic balance, place in a 100mL conical flask, add 9mL of purified water, and shake to mix.

[0037] After sampling the dairy products according to the above method, if there is turbidity, filter with 4 layers of filter paper, add 5mL of mixed acid, cover and place for 1h or overnight, place on an adjustable temperature-controlled electric heating furnace for digestion, wait for white smoke to appear, and the digestion liquid is colorless, transparent or slightly yellow, cool, dilute to 25mL with 0.01mol / L PBS (pH7.4) solution, mix well for standby use, take 100μL and test on the prepared test paper, and directly observe the C and T line after 10min. The detection limit for fresh milk / yogurt is 10μg / kg, the detection limit for finished milk is 30μg / kg, and the detection limit for milk powder / cheese is 300μg / kg.

[0038] 4. Vegetables, fruits, and aquatic products

[0039] Wash and dry high-water content samples such as vegetables (such as cucumbers, cabbages, etc.), fruits, and aquatic products, take the edible part and homogenize it evenly, filter it with 4 layers of filter paper, take 1.0g and add 5mL of mixed acid, cover it and place it for 1h or overnight, and place it on an adjustable temperature-controlled electric heating furnace for digestion. When white smoke appears and the digestion liquid is colorless and transparent or slightly yellow, cool it, dilute it to 25mL with 0.01mol / L PBS (pH7.4) solution, mix it for later use, take 100μL and test it on the prepared test paper, and directly observe the C and T lines after 10min. The detection limit is 1μg / kg.

[0040] 8. Edible Fungi

[0041] Wash the edible fungi, dry them, take the edible part, crush them with a grinder, pass through a 20-mesh sieve, take 1.0g, add 5mL of mixed acid, cover and place for 1h or overnight, place on an adjustable temperature-controlled electric heating furnace for digestion, wait until white smoke appears and the digestion liquid is colorless and transparent or slightly yellow, cool, dilute to 25mL with 0.01mol / L PBS (pH7.4) solution, filter with 4 layers of filter paper for later use, take 100μL and test on the prepared test paper, and directly observe the C and T lines after 10min. The detection limit is 1μg / kg.

[0042] 5. Seasoning (peppercorns, chili peppers, vinegar)

[0043] Take dried whole peppercorns, pepper peels and pepper seeds, crush them, pass through a 20-mesh sieve, accurately weigh 1.0g with an electronic balance, put them in a 100mL conical flask, add 5mL of mixed acid, cover and place for 1h or overnight, place on an adjustable temperature-controlled electric heating furnace for digestion, wait until white smoke appears and the digestion liquid is colorless and transparent or slightly yellow, cool, dilute to 25mL with 0.01mol / L PBS (pH7.4) solution, filter with 4 layers of filter paper for later use, take 100μL and test on the prepared test paper, and directly observe the C and T lines after 10min. The detection limit is 50μg / L.

[0044] Take pepper peel and seeds, wash with deionized water, oven dry, crush, accurately weigh 1.0g with an electronic balance, put into a 100mL conical flask, add 5mL of mixed acid, cover and place for 1h or overnight, place on an adjustable temperature-controlled electric heating furnace for digestion, wait until white smoke appears and the digestion liquid is colorless and transparent or slightly yellow, cool, dilute to 25mL with 0.01mol / L PBS (pH7.4) solution, mix well for use, take 100μL and test on the prepared test paper, and directly observe the C and T lines after 10min. The detection limit is 10μg / L.

[0045] Use a pipette to accurately draw 1.0mL of vinegar sample into a 100mL conical flask, add 5mL of mixed acid, cover and place for 1h or overnight, place on an adjustable temperature-controlled electric heating furnace for digestion, wait until white smoke appears and the digestion liquid is colorless and transparent or slightly yellow, cool, dilute to 25mL with 0.01mol / L PBS (pH7.4) solution, mix well for later use, take 100μL and test on the prepared test paper, and directly observe the C and T lines after 10min. The detection limit is 100μg / L.

[0046] 6. Liquor and distilled spirits

[0047] The sample is heated at low temperature on an electric furnace to remove ethanol, 1.0 mL is accurately pipetted into a 100 mL conical flask with a pipette, 5 mL of mixed acid is added, the mixture is placed on the cover for 1 hour or overnight, and then placed on an adjustable temperature-controlled electric furnace for digestion. When white smoke appears and the digestion liquid is colorless and transparent or slightly yellow, it is cooled and diluted to 25 mL with 0.01 mol / L PBS (pH 7.4) solution, mixed and set aside, 100 μL is taken for detection on the prepared test paper, and the C and T lines are directly observed after 10 minutes. The detection limit is 10 μg / L.

[0048] 7. Tea

[0049] The tea samples were evenly crushed by a high-speed grinder, passed through a 20-mesh sieve, dried in an 80°C drying oven to constant weight, and stored in a desiccator for later use.

[0050] Weigh 1g (accurate to 0.001g) tea sample accurately with analytical balance, put it in a 250mL conical flask, add 10mL mixed acid, cover and place for 1h or overnight, place on adjustable temperature control electric heating furnace for digestion, if it turns brown-black, add mixed acid again after cooling until white smoke appears, and the digestion liquid is colorless and transparent or slightly yellow, cool, dilute to 25mL with 0.01mol / L PBS (pH7.4) solution, filter with 4 layers of filter paper for later use, take 100μL and test on the prepared test paper, and directly observe the C and T lines after 10min. The detection limit is 100μg / g.

[0051] 8. Chinese herbal medicines and supplements

[0052] Wash and dry the low-water content samples such as Chinese herbal medicines and tonics, take the edible part, crush it with a grinder, pass it through a 20-mesh sieve, take 1.0g, add 5mL of mixed acid, cover it and place it for 1h or overnight, and place it on an adjustable temperature-controlled electric heating furnace for digestion. When white smoke appears and the digestion liquid is colorless and transparent or slightly yellow, cool it, dilute it to 25mL with 0.01mol / L PBS (pH7.4) solution, filter it with 4 layers of filter paper for later use, take 100μL and test it on the prepared test paper, and directly observe the C and T lines after 10min. The detection limit is 1μg / kg.

[0053] Take 1.0mL of Chinese medicine oral solution and add 5mL of mixed acid, cover and place for 1h or overnight, place on an adjustable temperature-controlled electric heating furnace for digestion, wait until white smoke appears and the digestion liquid is colorless and transparent or slightly yellow, cool, dilute to 25mL with 0.01mol / L PBS (pH7.4) solution, take 100μL and test on the prepared test paper, and directly observe the C and T lines after 10min. The detection limit is 1μg / kg.

[0054] [Example 3] Rapid detection of Mn using the colloidal gold test paper described in the utility model

[0055] Take a colloidal gold test paper preferably used in the present invention for rapid detection of Mn, such as Figure 1 As shown, it includes a supporting pad 1, a nitrocellulose membrane 2 located on the supporting pad 1, a Mn antibody Mn antibody detection line 3 coated in the middle of the nitrocellulose membrane 2, and control lines 4 respectively arranged on both sides of the Mn antibody detection line 3, the nitrocellulose membrane 2 is connected to a water absorbent pad 5 above one end, and connected to a conjugate pad 6 above the other end, and the conjugate pad 6 is connected to a sample pad 7; the Mn antibody detection line 3 is coated with monoclonal mouse anti-Mn antibodies, the conjugate pad 6 is coated with gold-labeled BSA-Mn and a control line conjugate, the control line 4 is coated with anti-DNP antibodies, and the control line conjugate is colloidal gold labeled with DNP-BSA.

[0056] The test was performed according to the following steps: 1) aspirate the sample to be tested, and if the sample was stored at low temperature, it needs to be restored to room temperature. 2) Add the sample to be tested to the sample pad 7 and react for 10 minutes. 3) Directly observe or place the colloidal gold test paper in a colloidal gold card reader (Guangdong Dayuan Oasis Food Safety Technology Co., Ltd.) to determine the result.

[0057] [Example 4] Rapid detection of Mn using the colloidal gold test paper box described in the utility model

[0058] Take a colloidal gold test paper box for rapid detection of Mn preferably used in the utility model, Figure 2As shown, it includes a support pad 1, a nitrocellulose membrane 2 located on the support pad 1, a Mn antibody detection line 3 coated in the middle of the nitrocellulose membrane 2 and control lines 4 respectively arranged on both sides of the Mn antibody detection line 3, the nitrocellulose membrane 2 is connected to a water-absorbing pad 5 above one end, and a conjugate pad 6 is connected to the other end, and the conjugate pad 6 is connected to a sample pad 7; the Mn antibody detection line 3 is coated with monoclonal mouse anti-Mn antibody, the conjugate pad 6 is coated with gold-labeled BSA-Mn and control line conjugate, the control line 4 is coated with anti-DNP antibody, and the control line conjugate is colloidal gold labeled with DNP-BSA. The colloidal gold test paper also includes a shell 9, which wraps the test paper, exposing the sample pad 7, the Mn antibody detection line 3 on the nitrocellulose membrane 2 and the two control lines 4. The test is performed according to the following steps: 1) The collected Mn sample to be tested, such as the sample stored at low temperature, needs to be restored to room temperature. 2) The sample to be tested is added to the sample pad 7 and reacted for 10 minutes. 3) directly observe or place the colloidal gold test paper in a colloidal gold card reader (Guangdong Dayuan Oasis Food Safety Technology Co., Ltd.) to determine the result.

Claims

1. A colloidal gold test paper for rapid detection of trace element manganese in food, comprising a support pad (1), a nitrocellulose membrane (2) located on the support pad (1), a Mn antibody detection line (3) coated in the middle of the nitrocellulose membrane (2), and control lines (4) respectively arranged on both sides of the Mn antibody detection line (3), wherein an absorbent pad (5) is connected to the top of one end of the nitrocellulose membrane (2), and a conjugate pad (6) is connected to the top of the other end, and the conjugate pad (6) is connected to the sample pad (7); It is characterized in that The Mn antibody detection line (3) is coated with monoclonal mouse anti-Mn antibody, and the conjugate gasket (6) is coated with gold-labeled BSA-Mn and control line conjugate.

2. Colloidal gold test paper as claimed in claim 1, It is characterized in that The control line (4) is coated with anti-DNP antibodies.

3. Colloidal gold test paper as claimed in claim 1, It is characterized in that A fixing pad (8) is provided above the sample pad (7).

4. Colloidal gold test paper as claimed in claim 3, It is characterized in that The fixing gasket (8) is a single-sided adhesive tape or a double-sided adhesive tape.

5. The colloidal gold test paper as claimed in claim 1, It is characterized in that The control line conjugate is colloidal gold labeled with DNP-BSA.

6. The colloidal gold test paper according to claim 1 or 3, It is characterized in that It also comprises a shell (9) wrapping the colloidal gold test paper, and is provided with windows respectively exposing the sample pad (7), the Mn antibody detection line (3) and the two control lines (4).

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