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Sample pretreatment liquid for detecting and separating respiratory viruses in samples

A pretreatment liquid and respiratory technology, applied in the direction of viruses/phages, microorganisms, biochemical equipment and methods, etc., can solve the problems of missing samples, wrong additions, and mismatching test results, etc.

Active Publication Date: 2016-07-13
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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  • Sample pretreatment liquid for detecting and separating respiratory viruses in samples

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] The preparation of embodiment 1 sample pretreatment liquid and its processing to sample (specimen)

[0062] 1. Components of the sample pretreatment solution

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

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

[0065] Component 3: solution containing 1ml of penicillin and streptomycin (the mother solution is penicillin 10,000 U / mL, 10,000 μg / mL of streptomycin is dissolved in 100ml of 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 (calculated 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 stir magnetically to help dissolve.

[0070] 2.3 Add component 3 to th...

Embodiment 2

[0077] The preparation of embodiment 2 sample pretreatment liquid and its processing to sample (specimen)

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

Embodiment 4

[0086] The impact 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 matter and a buffer system for respiratory viruses, which can better treat respiratory viruses. Viruses for protection.

[0088] The processed respiratory samples (Example 1, 2 and Comparative Example 1) were inoculated into growth state sensitive cell lines (dog kidney cell MDCK, rhesus monkey kidney cell LLC-MK2, human laryngeal cancer epithelial cell Hep-2, etc.), Place at 37°C, 5% CO 2 Culture in an incubator, and observe the cytopathic effect (cytopathic effect, CPE) every day. Cytopathy is characterized by swollen and rounded cells, increased intercellular spaces, and pyknotic or ruptured nuclei. In severe cases, some or all of the cells fall off. The change in CPE of 0-25% of cells is "+", and...

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PUM

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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 refers to the general term for viruses that mainly invade the respiratory tract, first proliferate in the epithelial cells of the respiratory tract mucosa, cause respiratory tract and systemic infection, and cause damage to the respiratory tract and other organs. 90-95% of clinical acute respiratory infections are caused by this group of viruses, the main respiratory viruses are influenza virus (A, B, C), parainfluenza virus (type 1, 2, 3, 4, 5) , respiratory syncytial virus, measles virus, mumps rhinovirus, coxsackie and echoviruses, coronaviruses, reoviruses (types 1, 2, 3, 4), adenoviruses (types 3, 4, 7 , 14, 21 types), human herpes virus rubella virus and cytomegalovirus, etc. The detection rate of respiratory viruses in lower respi...

Claims

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Application Information

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