Herpes simplex virus and recombinant virus as well as host cell and medicinal combination thereof
A technology of herpes simplex virus and recombinant virus, applied in the direction of drug combination, virus/bacteriophage, microorganism, etc., can solve the problems of slow replication and proliferation, low virus infection rate, etc., achieve fast replication, high specific infection rate, The effect of strong solvency
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Embodiment 1
[0195] This example illustrates the isolation of herpes simplex virus according to the invention.
[0196] (1) Collection of virus
[0197] An adult Chinese male, 37 years old, Han nationality, with a history of repeated oral herpes, without other diseases, had a typical clinical manifestation of herpes simplex virus type I recurrent infection—the sample was taken two days after herpes-like blisters appeared around the corner of the mouth.
[0198] Take a 15 ml sterile centrifuge tube, which contains 3 ml of sterile phosphate buffered saline (PBS) containing 100 μg / ml kanamycin, which contains 9 g / L of NaCl, 0.21 g / ml of liters of KH 2 PO 4 and 0.97 g / L of Na 2 HPO 4 .12H 2 O: Squeeze the herpetic vesicle with a sterile cotton swab until the light yellow secretion flows out, then dip the secretion with a cotton swab, and rinse the end of the obtained cotton swab with the secretion in the above-mentioned PBS buffer for 10 seconds. Take out the cotton swab, and use a steri...
Embodiment 2
[0206] This example illustrates the lytic ability of the herpes simplex virus of the present invention to human nasopharyngeal carcinoma cells (CNE-1) with multiple occurrences in yellow race.
[0207] (1) Determination of herpes simplex virus titer
[0208] Using DMEM medium containing 10% fetal bovine serum, in six-well cell culture plate at 5 × 10 5 Inoculate African green monkey kidney cells (Vero cells) (ATCC, USA) at a density of 1 cell / well, at 37°C in 5% CO 2 Cultivate in the incubator for 24 hours until each well of Vero cells is 80% full and ready for use.
[0209] Under sterile conditions, collect the supernatant culture solution of the Vero cell culture cultured with herpes simplex virus of the present invention in Example 1, and use serum-free DMEM medium to gradiently dilute to 1 × 10 respectively. 2 Double solution, 1×10 3 Double solution, 1×10 4 Double solution, 1×10 5 Double solution, 1×10 6 Double solution and 1×10 7 100 microliters of each dilution. ...
Embodiment 3
[0224] This example demonstrates the lytic ability of the herpes simplex virus of the present invention to normal tumor cells.
[0225] According to the method of Example 2, human breast cancer cells MCF-7, human colon cancer cells HT-29 and Human lung adenocarcinoma cell A549 (purchased from the Second Research Institute of the Academy of Military Medical Sciences), and the survival number of tumor cells at different time points after transfection were tested, and the results are shown in Table 3.
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