For the anti-virus efficacy test of new nano-composite materials, Bodochem Technology has conducted a coronavirus (SARS) test in Microbac, a laboratory designated by the FDA in the United States, and cooperated with Boken, the top three testing centers in Japan, to conduct experiments on influenza viruses and cooperate with stem cell clinical medicine. The research center cooperated with the influenza virus enterovirus test and passed the test. The above test verification can prove that the materials developed by Bo Medical Technology have excellent antiviral performance. The following is a detailed description of the various anti-virus tests and test results.

 

1. The United States FDA designated laboratory MicrobacInstructions for performing a coronavirus (SARS) test:
 

Summary:

Microbac laboratory conducts coronavirus (SARS) test, according to the international standard specification ASTM E1052-11 test method, the residual amount of the virus in the culture medium after the SARS virus interacts with the material.

 

Virus type:

A. Coronavirus: SARS-associated Coronavirus (CDC 200300592), from ZeptoMetrix/CDC virus strain.

B. Host cells: Vero E6 cell line (ATCC CRL-1586) was used.

 

Anti-virus test of new nano-composite materials:

This test is carried out in accordance with the ASTM E1052-11 test method. Under light conditions, the virus liquid without added materials and the virus liquid with added materials are cultured at the same time, and the virus infection situation is finally judged. The experimental test is divided into three parts: sample test, virus culture control and neutralizer effectiveness/virus interference and cytotoxicity, each of which is briefly described as follows:

 

1. Sample test: mix the test material with the virus liquid and irradiate it with a light source, add a neutralizer and then dilute it appropriately, and finally carry out the interpretation of the virus infection.

2. Virus culture control: mix the diluted culture solution with the virus solution, irradiate with a light source, add a neutralizer and then dilute appropriately, and finally carry out the interpretation of virus infection.

3. Neutralizer effectiveness/virus interference and cytotoxicity: Mix the test material with the diluted culture solution and irradiate it with a light source. After adding the neutralizer, dilute it appropriately, and divide it into two groups to confirm the neutralizer effectiveness/virus interference and Confirmation of cytotoxicity. Among them, the effectiveness of the neutralizer/virus interference is confirmed by adding a small amount of virus solution for culture; the cytotoxicity is directly interpreted without adding virus solution.

4. Calculate virus suppression ability = suppressed virus amount - inoculated virus amount.

5. The initial virus concentration value (Log10 TCID50) is 7.28, the material reduces the value of virus infection (Log10 TCID50) after 20 minutes of contact time is ≦3.61, and the value of reducing virus infection is ≧3.67.

 

2. Instructions for Influenza Virus Enterovirus Testing in cooperation with Taiwan Index Medical Research Center:

 

Summary:

Jingcheng Nano Technology cooperates with Stem Cell Clinical Medicine Research Center to conduct the following tests on the anti-virus of nano-composites: using the method of Plaque Forming in the transparent area,Observe the residual amount in the virus culture solution after the material is applied.

 

Virus type:

1. Enterovirus: Enterovirus Ico-11, from the American Pathology Society proficiency test virus strain.

2. Influenza A virus: Influenza A virus New Caledonia/20/99 (H1N1) comes from the ability test virus strain of the Society of American Pathology.

 

Host cell:

1. MDCK cell line (BCRC 60004) was used for type A influenza virus.

2. Enterovirus uses LLC-MK2 cell line (BCRC 60092).

 

Anti-virus test of new nano-composite materials

The virus types in this test are enterovirus and influenza A virus. When the cells are infected by the virus, the cells will die (as shown in the figure below), and then use the MTT crystal staining method to test the effect of the cell enzyme activity on the substrate. Analyze the cell survival rate to judge the effect of anti-virus. When the test material has a higher anti-virus effect, the higher the cell survival rate is. Since viruses and bacteria are different and cannot be directly measured, virus plaques are formed by cell infection ( Plaque) method, planCalculate virus plaque forming unit (PFU; Plaque-forming unit) as the judgment of virus concentration, and its unit is PFU/Ml.

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The above anti-virus tests are all in accordance with ASTM E 1052-96, the US FDA-certified laboratory MICROBIOTEST for anti-virus certification. According to this method, the test is carried out in cooperation with the Stem Cell Clinical Medical Research Center and the virus suppression ability is calculated = the amount of virus suppression ÷Volume of inoculated virus.

1. Tested at a virus concentration of 2×106 PFU/mL, the material has the effect of inhibiting enterovirus infection of LLC-MK2 cells, and the inhibition ability reaches 99.99%.

2. Tested at a virus concentration of 4×103 PFU/mL, the material has the effect of inhibiting type A virus from infecting MDCK cells, and the inhibition ability reaches 99.99%.

 

3. Boken-Osaka Microbiology Laboratory conducts influenza virus (H1N1) testing, and establishes a test method in accordance with the international standard specification ISO 21702 combined with the light conditions of JIS R 1702, and conducts the antiviral activity value after the H1N1 virus interacts with materials.

 

Virus type:

Influenza virus : Influenza A Virus (H1N1) (ATCC VR-1469).