Viscous biological sample digestive agent and preparation method thereof

A technology of biological samples and digestive agents, applied in the field of medical testing, can solve the problems of damage to pathogenic bacteria, low-temperature storage, and incompatibility, and achieve the effect of resisting pollution, acceptable cost, and not damaging pathogenic bacteria

Active Publication Date: 2021-01-26
珠海市德灏生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In summary, currently commonly used sample digestion reagents need to be stored at low temperature, need to be reconstituted by the user, are unstable in liquid state, damage pathogenic bacteria, and are not universal among different testing items

Method used

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  • Viscous biological sample digestive agent and preparation method thereof
  • Viscous biological sample digestive agent and preparation method thereof
  • Viscous biological sample digestive agent and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Embodiment 1 prepares experimental sample

[0049] One, prepare digestive agent solution of the present invention, get the component of following quality or volume and prepare:

[0050]

[0051] First dissolve the solutes other than papain to a constant volume, adjust the pH value to 7.2-7.4, then add papain, stir until completely dissolved, filter and sterilize through a microporous membrane with a pore size of 0.22um, use on the same day or refrigerate at 2-8°C for later use .

[0052] Two, the preparation of control sample trypsin liquid, its each component and consumption are as follows:

[0053]

[0054] First dissolve the solutes other than trypsin to a constant volume, and the measured pH value should be 7.8±0.1, then add trypsin, stir until completely dissolved, filter and sterilize through a microporous membrane with a pore size of 0.22um, use it on the same day or refrigerate at 2-8°C spare.

[0055] 3. Preparation of control sample saline

[0056] S...

Embodiment 2

[0059] Example 2 Experimental Sample Digestibility Test for Viscous Samples

[0060] 1. Sputum digestion test

[0061] 1) Take a clinical sputum sample, add 5ml of sterile saline, shake it several times, and absorb all the liquid around the sputum block;

[0062] 2) Repeat step 1) twice to obtain the experimental sputum block;

[0063] 3) The sputum block is divided into 3 equal parts, and the digestion experiment is carried out respectively with 3 kinds of experimental samples of Example 1 (digestive agent of the present invention, control sample trypsin solution, control sample physiological saline), and the amount of experimental samples added is as shown in Table 1 As shown (equal volume of sputum block and digestive juice added);

[0064] Table 1

[0065]

[0066] 4) Digestion, oscillate continuously, and observe the digestion effect of sputum block with naked eyes at 10 minutes, 20 minutes, and 30 minutes after adding the test solution.

[0067] Results: The digesti...

Embodiment 3

[0083] Embodiment 3 influence test on the culture detection of fastidious bacteria

[0084] Follow the steps below:

[0085] 1. Haemophilus influenzae strains recover and grow to logarithmic growth phase;

[0086] 2. Adjust the turbidity to 0.5MCF with sterile saline;

[0087] 3. Dilute 1:1000 with sterile normal saline, then divide into 2 equal parts, add equal volume of digestive agent solution of the present invention to one part as test group, add equal volume of normal saline to the other as control group, 37°C Constant temperature co-cultivation;

[0088] 4. At the time points of 10min, 30min, and 60min of co-incubation, take samples and make a 1:10 dilution with sterile saline, add 10ul to the chocolate plate medium, and draw the inoculation loop without dividing the line, so that it is evenly dispersed throughout the On the surface of the plate, three plates were repeated for each time point in each group;

[0089] 5. Cultivate and observe the number of colonies: C...

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Abstract

The invention discloses a viscous biological sample digestive agent composition. The viscous biological sample digestive agent composition comprises protease, a stabilizer and a synergist, wherein theprotease is papain, cathepsin, subtilisin or protease K; the stabilizer comprises saccharides, cyclodextrin and a biological buffer agent; and the synergist is sodium pyrophosphate, triethanolamine,sodium ethylene diamine tetracetate, tartaric acid or sodium citrate. The invention further provides a viscous biological sample digestive agent. The viscous biological sample digestive agent comprises any one of the digestive agent compositions and deionized water. The invention further provides a preparation method of the viscous biological sample digestive agent. The preparation method comprises the following steps of: dissolving the stabilizer, the synergist and a hypertonic agent in the deionized water, adjusting the pH value to 7.2-7.4, adding the protease, and stirring and dissolving toobtain the viscous biological sample digestive agent. The digestive agent solution is stable in liquid state, does not need to be compounded before being used by a user, does not damage pathogenic bacteria, can be stored at normal temperature, takes effect in the optimal pH range of the pathogenic bacteria, and can be universally used for different detection items.

Description

technical field [0001] The invention relates to the field of medical testing, in particular to a viscous biological sample digester and a preparation method thereof. Background technique [0002] Various clinical samples such as sputum and cervical samples are viscous. In these samples, the normal flora without clinical significance were mostly distributed on the surface of the mucus block, and the pathogenic bacteria with clinical significance were mostly wrapped inside the mucus block. When such samples are directly smeared, the formed components such as bacteria and cells overlap in multiple layers, which is difficult to stain and observe; when directly streaked and inoculated, the pathogenic bacteria inside the mucus block cannot be dispersed, and it is difficult to obtain a satisfactory separation effect. Therefore, before performing smear examination and isolation and culture examination on such specimens, the samples must be liquefied and homogenized, which is referr...

Claims

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Application Information

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IPC IPC(8): C12N9/50C12N9/96G01N1/28
CPCC12N9/50C12N9/96G01N1/28
Inventor 农高惠何林声
Owner 珠海市德灏生物科技有限公司
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