A method for detecting human parasitic diseases without using a glass slide and a cover glass

Abstract

This invention relates to the field of fecal testing, and more specifically, to a method for detecting human parasitic pathogens without the use of slides and coverslips, comprising the following steps: Step 1, adding an appropriate amount of water to the culture wells of a cell culture plate, placing an appropriate amount of feces in the water of the culture well and stirring evenly; Step 2, cutting a piece of textile or nylon mesh of similar size to the culture well and placing it into the culture well to isolate the fecal residue and fecal liquid from each other; Step 3, after stirring and mixing the fecal liquid, transferring the fecal liquid to the culture wells of a new cell culture plate, covering it with the cell plate cover, and examining it under a microscope using an inverted microscope.

Description

Technical Field

[0001] This invention relates to the field of fecal testing, and more specifically, to a method for detecting human parasitic pathogens without the use of slides and coverslips. Background Technology

[0002] The most commonly used fecal pathogen detection techniques include the direct smear method, the centrifugal sedimentation method, and the brine floation method.

[0003] 1. Direct smear method: Take a glass slide, put a drop of physiological saline on it, use a bamboo skewer or toothpick to take a piece of feces the size of millet or mung bean, stir it evenly in physiological saline, draw it into a circle with a diameter of about 1 cm, remove the fecal residue, cover with a coverslip, and examine it under a regular optical microscope.

[0004] 2. Centrifugation precipitation method: Take a 1.5 ml (15 ml) test tube, add 1 ml (10 ml) of distilled water, and use a bamboo skewer or toothpick to take a soybean-sized (1-2 g) sample of feces. Place the sample in the distilled water and stir well. Take the fecal residue, rotate at 1000-2000 rpm for 1-2 minutes, remove the supernatant, add 1 ml of distilled water, centrifuge to settle, and remove the supernatant. Repeat this process 3-4 times until the supernatant is clear. Remove the supernatant. Take 100-300 μL (1 ml) to resuspend the precipitate, take one drop with a pipette, place it on a glass slide, cover with a coverslip, and examine under a regular optical microscope.

[0005] 3. Saturated salt flotation method, also known as saturated salt flotation method, takes a flotation cup, adds 1 / 3 to 1 / 2 of saturated salt water, uses a bamboo skewer or toothpick to take a soybean-sized amount of feces, puts it in the saturated salt water and stirs it evenly, then adds more saturated salt water to near the mouth of the bottle, removes the fecal residue, and continues to add saturated salt water until the liquid level is slightly higher than the mouth of the bottle, covers the mouth of the bottle with a glass slide, lets it stand for 10-15 minutes, then quickly lifts the glass slide, flips it over and covers it with a coverslip, and examines it under a regular optical microscope.

[0006] All three detection methods require the use of slides and coverslips. However, using slides and coverslips presents several challenges: the possibility of fecal matter spilling onto the slides and contaminating the surrounding work surface and microscope; the difficulty in handling the slides; the risk of them falling during cleaning and clogging drains; and the possibility of injury from broken slides and coverslips. Furthermore, using slides and coverslips requires rapid microscopic examination by the examiner, otherwise the fecal matter will dry out, making it impossible to detect pathogens. Additionally, using slides and coverslips makes it difficult to collect pathogens found during the examination, especially after the fecal matter has dried. Collecting pathogens requires washing the slides with saline or water, which is cumbersome. Most unacceptable to examiners are the direct smear method, centrifugation sedimentation method, and saturated saline flotation method, as the examiner must endure the unpleasant odor of feces during microscopic examination.

[0007] Therefore, it is necessary to propose a method for detecting human parasitic pathogens without using slides and coverslips to solve the above problems. Summary of the Invention

[0008] To overcome at least one of the defects (deficiencies) of the prior art described above, the present invention provides a method for detecting human parasitic pathogens without using glass slides and coverslips.

[0009] To solve the above-mentioned technical problems, the technical solution of the present invention is as follows: A method for detecting human parasitic pathogens without using a slide and coverslip, comprising the following steps: Step 1: Add an appropriate amount of water to the culture wells of the cell culture plate, and place an appropriate amount of feces into the water in the culture well and stir well. Step 2: Cut a piece of textile or nylon mesh that is the same size as the culture well and place it into the culture well to separate the fecal residue and fecal liquid. Step 3: After stirring and mixing the fecal liquid, transfer it to the culture wells of a new cell culture plate, cover the plate with the cell plate cap, and examine it under a microscope using an inverted microscope.

[0010] Furthermore, the textile is gauze, and the nylon screen is a 50-mesh nylon screen or a 100-mesh nylon screen.

[0011] Furthermore, the cell culture plate is a 6-well cell culture plate, a 12-well cell culture plate, a 24-well cell culture plate, a 48-well cell culture plate, or a 96-well cell culture plate.

[0012] Furthermore, step 4 is included, in which the stirred fecal liquid is extracted and centrifuged, and the supernatant is discarded after centrifugation before the pathogens are collected.

[0013] Furthermore, in step 4, the centrifuge rotates at 1000-2000 revolutions per minute, and the centrifugation time is 1-2 minutes.

[0014] Furthermore, in step 4, pathogens are sampled through the sampling groove on the sampling rod. In use, in addition to obtaining a positive result by taking an appropriate amount of sample, a quantitative result can also be obtained.

[0015] Compared with the prior art, the beneficial effects of the technical solution of the present invention are: The present invention discloses a method for detecting human parasitic pathogens without the use of slides and coverslips. When it is necessary to collect pathogens, the pathogens can be collected by centrifuging with a plate centrifuge or by using a pipette to draw fecal fluid into a 1.5ml centrifuge tube and centrifuging. During centrifugation, the speed is controlled at 1000-2000 rpm for 1-2 minutes, and the supernatant is discarded. The pathogens can then be collected. This method is simple and convenient. Attached Figure Description

[0016] Figure 1 This is a flowchart of the method for detecting human parasitic pathogens without using glass slides and coverslips, as described in this invention. Detailed Implementation

[0017] The accompanying drawings are for illustrative purposes only and should not be construed as limiting the scope of this patent. To better illustrate this embodiment, some components in the drawings may be omitted, enlarged, or reduced, and do not represent the actual dimensions of the product. It is understandable to those skilled in the art that some well-known structures and their descriptions may be omitted in the drawings.

[0018] In the description of this invention, it should be noted that, unless otherwise explicitly specified and limited, the terms "installation" and "connection" should be interpreted broadly. For example, they can refer to a fixed connection, a detachable connection, or an integral connection; they can refer to a mechanical connection or an electrical connection; they can refer to a direct connection or an indirect connection through an intermediate medium; they can refer to the internal communication between two components. Those skilled in the art can understand the specific meaning of the above terms in this invention based on the specific circumstances. The technical solution of this invention will be further described below with reference to the accompanying drawings and embodiments.

[0019] like Figure 1 As shown, a method for detecting human parasitic pathogens without using slides and coverslips includes the following steps: Step 1: Add an appropriate amount of water to the culture wells of the cell culture plate, and place an appropriate amount of feces into the water in the culture well and stir well. Step 2: Cut a piece of textile or nylon mesh that is the same size as the culture well and place it into the culture well to separate the fecal residue and fecal liquid. Step 3: After stirring and mixing the fecal liquid, transfer it to the culture wells of a new cell culture plate. After covering the cell plate with the cap, examine it under a microscope using an inverted microscope. This setup is to prevent contamination. In practical applications, sealing film can also be used to seal the cell plate cap.

[0020] In this invention, the textile is gauze, the nylon screen is a 50-mesh nylon screen or a 100-mesh nylon screen, and the cell culture plate is a 6-well cell culture plate or a 12-well cell culture plate or a 24-well cell culture plate or a 48-well cell culture plate or a 96-well cell culture plate.

[0021] In addition, step 4 is included, in which the stirred fecal liquid is extracted and centrifuged. After centrifugation, the supernatant is discarded and the pathogens are collected. In step 4, the centrifuge speed is 1000-2000 rpm and the centrifugation time is 1-2 minutes. In step 4, the pathogens are sampled through the sampling groove on the sampling rod.

[0022] The positional relationships described in the figures are for illustrative purposes only and should not be construed as limiting this patent. Clearly, the above embodiments of the present invention are merely examples to clearly illustrate the invention and are not intended to limit the implementation of the invention. Those skilled in the art can make other variations or modifications based on the above description. It is neither necessary nor possible to exhaustively list all possible implementations here. Any modifications, equivalent substitutions, and improvements made within the spirit and principles of the present invention should be included within the scope of protection of the claims of this invention.

Claims

1. A method for detecting human parasitic pathogens without using slides and coverslips, characterized in that: Includes the following steps: Step 1: Add an appropriate amount of water to the culture wells of the cell culture plate, and place an appropriate amount of feces into the water in the culture well and stir well. Step 2: Cut a piece of textile or nylon mesh that is the same size as the culture well and place it into the culture well to separate the fecal residue and fecal liquid. Step 3: After stirring and mixing the fecal liquid, transfer it to the culture wells of a new cell culture plate, cover the plate with the cell plate cap, and examine it under a microscope using an inverted microscope.

2. The method for detecting human parasitic pathogens without using slides and coverslips according to claim 1, characterized in that: The textile is gauze, and the nylon screen is a 50-mesh nylon screen or a 100-mesh nylon screen.

3. The method for detecting human parasitic pathogens without using slides and coverslips according to claim 1, characterized in that: The cell culture plate is a 6-well cell culture plate, a 12-well cell culture plate, a 24-well cell culture plate, a 48-well cell culture plate, or a 96-well cell culture plate.

4. The method for detecting human parasitic pathogens without using slides and coverslips according to claim 1, characterized in that: It also includes step 4, which involves extracting the stirred fecal liquid and centrifuging it. After centrifugation, the supernatant is discarded and the pathogens are collected.

5. The method for detecting human parasitic pathogens without using slides and coverslips according to claim 4, characterized in that: In step 4, the centrifuge rotates at 1000-2000 revolutions per minute, and the centrifugation time is 1-2 minutes.

6. The method for detecting human parasitic pathogens without using slides and coverslips according to claim 4, characterized in that: In step 4, pathogens are sampled through the sampling slot on the sampling rod.

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