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Large-scale cultivation method of human parainfluenza virus I

A technology of large-scale culture and virus culture, applied in the field of large-scale culture of human parainfluenza type I virus, can solve the problems of affecting the research of human parainfluenza type I virus vaccine, long culture period, inconspicuous lesions, etc., and achieves reduction of cell preparation. Quantity, scale expansion, and good cell viability

Inactive Publication Date: 2018-11-27
ZHENGZHOU IMMUNO BIOTECH
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  • Claims
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Problems solved by technology

Due to the inconspicuous lesions of human parainfluenza type I virus, slow toxin production, long culture period and high cost, the further research on human parainfluenza type I virus vaccine has been seriously affected

Method used

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  • Large-scale cultivation method of human parainfluenza virus I

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Embodiment Construction

[0013] The method of the present invention will be described in more detail below through specific examples, so that those skilled in the art can understand the present application.

[0014] The method for large-scale cultivation of human parainfluenza type I virus of the present invention comprises the steps::

[0015] In the first step, the human parainfluenza type I virus was rapidly thawed at 37°C, and the virus inoculation volume was calculated according to 0.01MOI, and the required volume of virus seed was inoculated on Vero cells covered with a single layer, and supplemented with 2% Calf serum virus maintenance solution, at 37 ° C, 5% CO 2 to cultivate;

[0016] In the second step, on the first day after inoculation, aseptically extract the culture supernatant, and use the ELISA sandwich method to detect the virus titer in the supernatant, and then operate synchronously every day to draw the virus proliferation curve. The titer test was carried out on the 7th day, and...

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Abstract

The invention discloses a large-scale cultivation method of human parainfluenza virus I, comprising: inoculating human parainfluenza virus I to Vero cells grown in a single layer according to 0.001-0.01 MOI; cultivating under 37 DEG C and 5% CO2; detecting viral titer in supernate each day by means of ELISA (enzyme-linked immuno sorbent assay), and drawing a viral proliferation curve; on seventh day, collecting viral culture supernate, and using a pancreatin digestive solution to digest cells, carrying out passage amplification of virus-carrying cells in the ratio of 1:(4-5), and culturing under 37 DEG C and 5%CO2; repeatedly collecting the viral solution, and adding the viral culture solution, or repeatedly carrying out passage amplification of virus-carrying cells until a required scale.Passage amplification of virus-carrying cells is carried out, the viral titer is high, cell survival rate is high, cell preparation quantity is decreased, cultivation time is shortened, and working efficiency is improved; in addition, passage amplification of the virus-carrying cells is carried out in the ratio of 1:(4-5) with no need for secondary virus inoculation; the scale can be widened as required, and the requirement for deep research and manufacture of human parainfluenza virus I vaccines is met.

Description

technical field [0001] The invention relates to a method for cultivating viruses, in particular to a method for large-scale culturing of human parainfluenza type I virus. Background technique [0002] Human parainfluenza viruses (HPIVs) are viruses that cause lower respiratory tract infections in children, and their pathogenicity is second only to respiratory syncytial virus (RSV). Like RSV, human parainfluenza viruses can cause recurrent upper respiratory infections (such as colds and sore throats), but they can also cause severe recurrent lower respiratory illnesses (such as pneumonia, bronchitis, and bronchiolitis), especially It is more prevalent in the elderly and in immunocompromised populations. [0003] Human parainfluenza type I viruses are single-stranded RNA viruses. The surface of the virus contains glycoprotein spines of lysozyme and hemagglutinin-neuraminidase. Serologically, human parainfluenza viruses can be divided into type I, type II, type III, and type...

Claims

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Application Information

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IPC IPC(8): C12N7/00
CPCC12N7/00C12N2760/18651
Inventor 周伟伟周雷鸣孙静静赵巧辉李桂林付光宇吴学炜
Owner ZHENGZHOU IMMUNO BIOTECH
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