Method for detecting nematode eggs by NaHCO3 saturated salt water flotation polymerization method
By using NaHCO3 instead of NaNO3 in the saturated saline floating pooling method, the problem of difficulty in obtaining NaNO3 is solved, effective detection of nematode eggs is achieved, and detection quality and public safety are ensured.
Patent Information
- Application Number
- CN202411147951.1
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-08-21
- Publication Date
- 2025-05-06
AI Technical Summary
In the prior art, the classic saturated saline floating pooling method is difficult to obtain normally, resulting in difficulties in detecting nematode eggs and micromembrane shell tapeworm eggs.
NaHCO3 was used to replace NaNO3, and nematode eggs were detected by preparing saturated saline and adding saturated NaHCO3 in combination with floating polymerization method. The method includes the preparation of saturated saline, the addition of the substance to be tested and the mashed thoroughly, the addition of saturated NaHCO3 and the removal of the coarse residue, the placement and the detection specimen is finally carried out under a microscope.
By replacing NaNO3 with NaHCO3 to ensure detection quality and public safety, it provides an easy-to-procure alternative reagent, and does not cause damage to nematode eggs, which can make nematode eggs rise and gather normally, solving the problem of difficulty in obtaining NaNO3.
Abstract
Description
Technical Field
[0001] The invention relates to the technical field of worm egg and microhymenolepithecidae egg detection, in particular to a method for detecting nematode eggs by using a NaHCO3 saturated salt water flotation method. Background Art
[0002] The classic saturated brine flotation method uses supersaturated NaNO3, but because it is one of the main raw materials for manufacturing nitroglycerin explosives, small-scale purchases also need to be reviewed and approved by the public security department. However, if local areas detect nematode eggs and microscopic tapeworm eggs, it is usually a project work, and the purchase volume at a time will be very large, which brings huge challenges to procurement and security work. For public safety considerations, the public security organs cannot approve such a large-scale purchase. Since the public security organs began to pre-approval the purchase of narcotics, easily made drugs, and easily made explosives, this reagent is almost impossible to buy in chemical reagent stores, so finding alternative reagents is currently a hot issue that the entire industry is paying attention to.
[0003] The density of the floating liquid is an important factor affecting the detection rate of Ascaris eggs. Using a solution with a high density as the floating liquid is more ideal. Compared with saturated zinc sulfate, magnesium sulfate, sodium chloride and other salt solutions, saturated sodium nitrate has the highest density and has the highest detection rate of Ascaris eggs when used as a floating liquid. In the absence of sodium nitrate, the above-mentioned saturated salt solutions can also be used instead. Among them, saturated sodium chloride solution has a poor floating effect on unfertilized Ascaris eggs.
[0004] In actual work, NaHCO3 is chosen to replace NaNO3. The molecular weight of the former is 84, and the molecular weight of the latter is 85. The particles contained in both are essential components for the normal growth of organisms that are widely present in nature. They will not cause damage to nematode eggs and H. microglossum eggs, nor will they cause the death of live eggs. Therefore, the safety significance of the replacement is obvious, and it will not cause any adverse effects on the detection of nematode eggs and H. microglossum eggs.
[0005] Therefore, how to provide a method for detecting nematode eggs by the NaHCO3 saturated salt water flotation method has become a technical problem to be solved by those skilled in the art. Summary of the invention
[0006] The technical problem to be solved by the present invention is how to provide a method for detecting nematode eggs by using a NaHCO3 saturated salt water flotation method.
[0007] To achieve the above object, the present invention provides a method for detecting nematode eggs by using a NaHCO3 saturated salt water flotation method, comprising the following steps:
[0008] S1: Preparation of saturated brine;
[0009] S2: Pick up the object to be tested and put it into the test container, add the saturated salt water prepared above, and fully mash and mix;
[0010] S3: Add saturated NaHCO3 salt water to the test container near the bottle mouth, and pick out the coarse residue with large particles;
[0011] S4: Use a dropper to add saturated salt water until the liquid level is slightly higher than the mouth of the test container;
[0012] S5: Take a clean glass slide and gently place it on the liquid surface of the test container bottle mouth to let it stand, and take the test specimen;
[0013] S6: After standing, place the slide under a microscope for inspection.
[0014] Preferably, when preparing saturated brine, 400 g of NaHCO 3 is slowly added into a container containing 1000 ml of boiling water and stirred for 3-5 min. After stirring, the mixture is cooled and the supernatant is taken for use.
[0015] During the slow addition of NaHCO3, stir continuously using a laboratory stirrer until NaHCO3 no longer dissolves.
[0016] Preferably, the object to be detected is one of feces or soil, and the radius is 4-5 mm.
[0017] The object to be detected with a radius of 4-5 mm is the size of a soybean. During the detection, a bamboo stick is used to pick up the object to be detected and put it into the detection container. The cost of using the bamboo stick is relatively low.
[0018] Preferably, the detection container is a flotation bottle with a height of 3.5 cm and a diameter of 2 cm.
[0019] The flotation bottle adopts the existing structure in the market in this field. The design of the flotation bottle uses a liquid with a large specific gravity through the flotation method to make the protozoan cysts or worm eggs in the object to be detected float and concentrate on the surface of the liquid for easy sampling and testing.
[0020] Preferably, the detection container is one of a penicillin bottle or a beaker.
[0021] Penicillin bottles or beakers can replace the floating bottles as detection containers, and both penicillin bottles and beakers adopt existing structures on the market, wherein the penicillin bottle is a large structure in the laboratory, and the beaker is a small structure in the laboratory.
[0022] Preferably, the standing time is 20 min.
[0023] When standing still, it should be in a clean state of dryness and no oil stains. When standing still, slowly cover the water droplets with a coverslip from one side to prevent bubbles. After standing still, lift it vertically and quickly and steadily turn the slide over, and place it under an external microscope for observation and inspection. When covering with a slide, you can also let it stand for 20 minutes before covering it. At this time, the covered dry slide can immediately contact the insect eggs floating on the liquid surface, with strong adhesion and better effect.
[0024] The beneficial effects of the present invention are:
[0025] When the present invention is used, the reagent replacement does not have any adverse effect on the test results under the premise of ensuring the quality of the test and public safety, and the reagents used are easy to purchase, and the amount of the substance is almost the same, NaHCO3 replaces NaNO3, and Na + 、HCO3 - 、NO3 - They are all common particles that maintain the normal growth of organisms and do not cause damage to nematode eggs. Supersaturated salt water can make the nematode eggs float and aggregate normally, which completely solves the dilemma that NaNO3 used in the classic saturated salt water flotation method can no longer be obtained normally but is needed for clinical and public health, thus facilitating its promotion and use. DETAILED DESCRIPTION
[0026] The embodiments of the present invention are described in detail below, and preferably the same or similar reference numerals throughout represent the same or similar elements or elements with the same or similar functions. The embodiments are exemplary and are intended to be used to explain the present invention, and should not be construed as limiting the present invention.
[0027] In the description of the present invention, it should be understood that the terms "length", "width", "up", "down", "front", "back", "left", "right", "vertical", "horizontal", "top", "bottom", "inside", "outside" and the like indicate positions or positional relationships, which are only for the convenience of describing the present invention and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, be constructed and operate in a specific orientation, and therefore cannot be understood as limiting the present invention. In addition, in the description of the present invention, the meaning of "multiple" is two or more, unless otherwise clearly and specifically defined.
[0028] The present invention provides a method for detecting nematode eggs by using a NaHCO3 saturated salt water flotation method, comprising the following steps:
[0029] S1: Preparation of saturated brine;
[0030] S2: Pick up the object to be tested and put it into the test container, add the saturated salt water prepared above, and fully mash and mix;
[0031] S3: Add saturated NaHCO3 salt water to the test container near the bottle mouth, and pick out the coarse residue with large particles;
[0032] S4: Use a dropper to add saturated salt water until the liquid level is slightly higher than the mouth of the test container;
[0033] S5: Take a clean glass slide and gently place it on the liquid surface of the test container bottle mouth to let it stand, and take the test specimen;
[0034] S6: After standing, place the slide under a microscope for inspection.
[0035] When preparing saturated brine, slowly add 400g of NaHCO3 into a container containing 1000ml of boiling water and stir for 3-5min. Cool after stirring. Take the supernatant after cooling for use. The object to be detected is one of feces or soil, and the radius is 4-5mm. The detection container is a flotation bottle with a height of 3.5cm and a diameter of 2cm. The detection container is one of the penicillin bottle or beaker. The standing time is 20min. Specific embodiment one:
[0037] In this embodiment:
[0038] First, prepare an appropriate amount of saturated brine according to demand;
[0039] Secondly, use a bamboo stick to pick up 1g of feces and put it into a flotation bottle, add an appropriate amount of the saturated salt water prepared above, and use a bamboo stick to mash the feces and saturated salt water to mix well;
[0040] Next, add saturated NaHCO3 salt water to the mouth of the floatation bottle, and use a bamboo stick to pick out the coarse residue with large particles;
[0041] Next, add saturated salt water with a dropper until the liquid level in the float bottle is slightly higher than the bottle mouth, until it stops overflowing;
[0042] Then, take a clean glass slide and gently place it on the liquid surface of the floating bottle. When placing the glass slide, be careful not to generate bubbles. After placing it, let it stand for 20 minutes.
[0043] Finally, when the resting is complete, lift the slide vertically and flip it over quickly and smoothly, and place it under an external microscope for observation and testing. Specific embodiment 2:
[0045] In this embodiment:
[0046] First, prepare an appropriate amount of saturated brine according to demand;
[0047] Secondly, use a bamboo stick to pick up 1g of soil and put it into a beaker, add an appropriate amount of the saturated salt water prepared above, and use a bamboo stick to mash the soil and saturated salt water to mix well;
[0048] Next, add saturated NaHCO3 salt water to the beaker near the bottle mouth, and use a bamboo stick to pick out the coarse residue with large particles;
[0049] Next, use a dropper to add saturated salt water until the liquid level in the beaker is slightly higher than the bottle mouth, until it stops overflowing;
[0050] Then, let it stand for 20 minutes;
[0051] Finally, when the standing is completed, take a clean glass slide and gently place it on the liquid surface of the beaker to take samples. After the sampling is completed, lift the glass slide vertically and turn it over quickly and smoothly, and place it under an external microscope for observation and inspection.
[0052] What is disclosed above is only a preferred embodiment of the present invention, and it certainly cannot be used to limit the scope of rights of the present invention. Ordinary technicians in this field can understand that all or part of the processes of the above embodiment and equivalent changes made according to the claims of the present invention still fall within the scope of the invention.
Claims
1. A method for detecting nematode eggs by NaHCO3 saturated salt water flotation method, comprising the following steps: S1: Preparation of saturated brine; S2: Pick up the object to be tested and put it into the test container, add the saturated salt water prepared above, and fully mash and mix; S3: Add saturated NaHCO3 salt water to the test container near the bottle mouth, and pick out the coarse residue with large particles; S4: Use a dropper to add saturated salt water until the liquid level is slightly higher than the mouth of the test container; S5: Take a clean glass slide and gently place it on the liquid surface of the test container bottle mouth to let it stand, and take the test specimen; S6: After standing, place the slide under a microscope for inspection.
2. The method for detecting nematode eggs by NaHCO3 saturated salt water flotation method as claimed in claim 1, characterized in that: When preparing saturated brine, slowly add 400g of NaHCO3 into a container containing 1000ml of boiling water and stir for 3-5min. After stirring, cool it and take the supernatant for use.
3. The method for detecting nematode eggs by NaHCO3 saturated salt water flotation method as claimed in claim 1, characterized in that: The object to be detected is one of feces or soil, and the radius is 4-5mm.
4. The method for detecting nematode eggs by NaHCO3 saturated salt water flotation method as claimed in claim 1, characterized in that: The detection container is a floating bottle with a height of 3.5 cm and a diameter of 2 cm.
5. The method for detecting nematode eggs by NaHCO3 saturated salt water flotation method as claimed in claim 4, characterized in that: The test container is either a penicillin bottle or a beaker.
6. The method for detecting nematode eggs by NaHCO3 saturated salt water flotation method as claimed in claim 1, characterized in that: The standing time is 20 minutes.