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A sample pretreatment solution for detecting and separating respiratory virus from samples

A pre-treatment liquid and respiratory technology, applied in the direction of virus/bacteriophage, microorganism, biochemical equipment and methods, etc., can solve the problems of missed addition of samples, mismatched test results, wrong addition, etc.

Active Publication Date: 2020-05-26
SHANGHAI PUDONG NEW AREA CENT FOR DISEASE CONTROL & PREVENTION
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the actual detection work, since the sample and the sample processing solution are colorless or white, when adding the sample, the color of the sample is consistent with the color of the sample plate or forms a transparent shape, which is easy to cause missing or wrong addition of the sample. resulting in discrepancies in the test results

Method used

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  • A sample pretreatment solution for detecting and separating respiratory virus from samples

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] Example 1 Preparation of sample pretreatment solution and its treatment of samples (specimen)

[0062] 1. Composition of sample pretreatment solution

[0063] Component 1: contains 200 mg of calcium chloride; 97.67 mg of anhydrous magnesium sulfate; 400 mg of potassium chloride; 6800 mg of sodium chloride; 122 mg of anhydrous sodium dihydrogen phosphate and 1000 mg of anhydrous glucose.

[0064] Component 2: Contains 10 mg of phenol red.

[0065] Component 3: 1ml penicillin-streptomycin solution (the mother solution is penicillin 10,000U / mL, 10,000μg / mL streptomycin dissolved in 100ml 0.85% NaCl).

[0066] Component 4: Contains 4766 mg of 2-[4-(hydroxyethyl)-1-piperazinyl]ethanesulfonic acid.

[0067] 2. Preparation steps of sample pretreatment solution (in 1L)

[0068] 2.1 Pour 50mL of distilled water into a clean glass.

[0069] 2.2 Pour components 1-2 into a glass, and magnetically stir to help dissolve.

[0070] 2.3 Add component 3 to the glass.

[0071] 2.4 A...

Embodiment 2

[0077] Example 2 Preparation of sample pretreatment solution and its treatment of sample (specimen)

[0078] In this example, the composition of the sample pretreatment solution was 10 mM PBS, no phenol red was added, and the pH was 7.4. Using the method of Example 1, lower respiratory tract specimens were processed.

Embodiment 4

[0086] The effect of embodiment 4 on virus isolation

[0087] Since the main components of the sample pretreatment solution in Example 1 are close to the composition of the virus separation solution, the sample pretreatment solution in Example 1 provides inorganic salts, organic substances and a buffer system for respiratory viruses, which can better protect the respiratory tract. virus protection.

[0088] The treated respiratory samples (Examples 1, 2 and Comparative Example 1) were inoculated into growth-state sensitive cell lines (dog kidney cells MDCK, Rhesus monkey kidney cells LLC-MK2, human laryngeal carcinoma epithelial cells Hep-2, etc.), Place at 37°C, 5% CO 2 The cells were cultured in an incubator, and the cytopathic effect (CPE) was observed daily. Cytopathy is characterized by swelling and rounding of cells, enlargement of intercellular spaces, pyknosis or rupture of nuclei, and partial or complete shedding of cells in severe cases. "++", 51-75% cells CPE cha...

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Abstract

The invention provides a sample pretreatment liquid for detecting and separating respiratory viruses in samples.Particularly, the sample pretreatment liquid with a pH value of 7.2-7..8 comprises (i) a virus preserver which is selected from calcium chloride, anhydrous magnesium sulfate, potassium chloride, sodium chloride, sodium dihydrogen phosphate anhydrous, anhydrous glucose or a combination thereof, (ii) a color developing agent, (iii) a sterilizing agent comprising penicillin, streptomycin or penicillin-streptomycin and (iv) a pH regulator.When used for treating the samples, the sample pretreatment liquid is capable of decreasing sample adding error rate remarkably, keeping virus activity and improving separated virus expression efficiency.

Description

technical field [0001] The invention relates to the field of biological detection, in particular, to a sample pretreatment solution for detecting and separating respiratory viruses in samples. Background technique [0002] Respiratory virus is a general term for viruses that mainly use the respiratory tract as an invasion portal, and first proliferate in the epithelial cells of the respiratory tract, causing respiratory and systemic infections, and causing damage to the respiratory tract and other organs. 90 to 95% of clinical acute respiratory infections are caused by this group of viruses. The main respiratory viruses are influenza virus (A, B, C) and parainfluenza virus (types 1, 2, 3, 4, and 5). , respiratory syncytial virus, measles virus, parotid rhinovirus, coxsackie virus and echo virus, coronavirus, reovirus (types 1, 2, 3, 4), adenovirus (3, 4, 7 , 14, 21 type), human herpes virus rubella virus and cytomegalovirus. The detection rate of respiratory viruses in low...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N7/00
CPCC12N7/00
Inventor 潘丽峰
Owner SHANGHAI PUDONG NEW AREA CENT FOR DISEASE CONTROL & PREVENTION