4-Hydroxyethylpiperazine ethanesulfonic acid, English abbreviation HEPES, CAS number: 7365-45-9, color is white crystalline powder, PH value range is 6.8-8.2, its most common application is as a biological buffer adjustment PH value and its application in skin care products are also loved by major manufacturers. Its application in rabies virus detection is often unknown. Today, the editor of Desheng will popularize relevant knowledge for everyone.
Rabies virus generally does not produce cytopathic (CPE) when it reproduces in infected cells, and it is not easy to form plaques under conventional culture conditions. Although many scholars at home and abroad have established rabies virus erosion on chicken embryo cells and BHK-21 cells. Spot methods, but these methods require strict experimental conditions, or the operations are complicated and difficult to master, which affects their popularization and use. After the researchers found that 4-hydroxyethylpiperazine ethanesulfonic acid HEPES can enhance the pathogenic effect of rabies virus on infected cells, they used this characteristic of HEPES to establish a simple and rapid HEPES plaque method for rabies virus.
The effect of HEPES on the formation of rabies virus plaque
After the infected cells were cultured at 37°C for 3 to 5 days, the infected cells treated with HEPES developed obvious cytopathic changes at the bottom of the covering. After staining with crystal violet, clear plaque formation was seen, while the infected cell group not treated with HEPES showed no signs of plaque formation. There is no cytopathic effect, and no plaque formation. It can be seen that HEPES can obviously promote the formation of rabies virus plaque on BHK cells.
Observation on the sensitivity of HEPES plaque method
The HEPES plaque method can be used for the titer of virus infection. Experiments show that there is an obvious titer relationship between the number of plaques formed after virus infection and virus dilution. The infective titers of the same rabies virus CTN-BHK strain were measured by the HEPES plaque method and the mouse method. The results showed that the number of plaques obtained by the HEPES plaque method for 4 consecutive determinations ranged from 1.0×10° to 4.0× 10*PFU/m1. The infection titer obtained by the mouse method for 4 consecutive determinations is 6.0～6.8log or 1.0×10°～6.3×10/ml. This shows that the rapid HEPES plaque method has the same sensitivity as the mouse method.
Advantages of HEPES plaque method
Using rabies virus CTN-181 strain and CTN-BNK strain to study the infection titer titration method, the results show that HEPES can induce and enhance the pathogenic effect of the canine virus on the infected cells. When the virus infects the cells, they are cultured at 37°C for 3～5 Just add 50～00mmol/L HEPES on the cover of methylcellulose semi-solid medium on the first day, stain with crystal violet solution after 24 hours, and calculate the number of plaques by naked eyes after drying at room temperature. Compared with the insect spot method reported in the previous literature, this plaque method has the advantages of fast, simple, economical, sensitive and easy to master.
The HEPES produced by Desheng is reagent grade with a purity greater than 99%. Our products perform well. There are many cooperative manufacturers and can provide samples for free trial. Desheng will continue to provide customers with high-quality products as always. If you have any questions or needs, you can go to the official website to consult customer service staff
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