Removal method of residual disinfectant in virus inactivation test

A virus inactivation and disinfectant technology, applied in the field of disinfection, can solve the problems that the neutralizer components cannot be harmful to microorganisms or cells, the chemical neutralizer is difficult to find, and the cells are affected, so as to eliminate adverse effects, reduce test time, The effect of improving detection efficiency

Inactive Publication Date: 2020-01-03
GUANGDONG PROVINCIAL INST OF BIOLOGICAL PROD & MATERIA MEDICA +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, the removal methods of residual disinfectants mainly include chemical neutralization and physical methods. The chemical neutralization method uses the antagonism between chemical substances to eliminate residual disinfectants, but different chemical neutralizers and neutralized products in this method can affect the cells
It is necessary to remove the residual disinfectant in each test. The method needs to be qualified before it can be applied, but the neutralizer method has considerable shortcomings: 1. The screening of compound disinfectant neutralizer components is very cumbersome and the success rate is very low; 2. Different The concentration of disinfectant effective substance is inconsistent, and the concentration of commonly used neutralizer components is not necessarily reasonable; 3. The neutralizer components themselves cannot be harmful to microorganisms or cells
Therefore, the chemical neutralization method needs to carry out a sufficient amount of preliminary tests, which consumes a lot of time and manpower, and as the composition of the disinfectant becomes more and more complex, it is more and more difficult to find a suitable chemical neutralizer, which has become a key factor in the virus inactivation test. Difficulties: Physical methods include dilution method, adsorption column method, molecular sieve column method and carrier washing method. These methods have special requirements for disinfectants or viruses and cannot be universally promoted

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  • Removal method of residual disinfectant in virus inactivation test
  • Removal method of residual disinfectant in virus inactivation test
  • Removal method of residual disinfectant in virus inactivation test

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specific Embodiment approach

[0023] A method for removing residual disinfectant in a virus inactivation test, characterized in that said method for removing comprises the following steps:

[0024] 1) Preparation steps: prepare virus suspension; dilute the disinfectant to be tested with sterilized hard water to 1.25 times the concentration used;

[0025] 2) Removal steps: first absorb the antiseptic into the test tube, perform antiseptic treatment after water bath, then perform 30-50 times dilution and then ultracentrifuge, then discard the supernatant, and finally add cell maintenance solution to dilute and mix to remove residual Disinfectant; in this embodiment, add cell maintenance solution and dilute and mix the volume to 1mL~2mL, the volume ratio of virus suspension to disinfectant is 1:4, and the virus suspension is 0.2mL~0.4mL; where the water bath temperature The temperature is 20°C±1°C, the time is 5min, the centrifugation conditions are: speed 10000rpm-25000rpm, centrifugation time 1h-2.5h, tempe...

specific Embodiment

[0034] 1. Scope of application

[0035] This embodiment is mainly applicable to disinfection product identification or daily monitoring, with a certain representative, live virus and its cell infection technology, to evaluate the killing effect of various disinfection factors on the test virus, to the inactivation of disinfection factors Important aspects of viral competence were validated.

[0036] 2. Test equipment

[0037] (1) Virus strains used in the test: poliovirus type Ⅰ (poliovirus-Ⅰ, PV-Ⅰ) vaccine strain, enterovirus type 71 (EV71) provided by the Guangdong Provincial Center for Disease Control and Prevention;

[0038] (2) Host cells: Hep-2 cells, as test cells for PV-1; VERO cell lines, as test cells for EV71; both cells were provided by Guangdong Provincial Center for Disease Control and Prevention;

[0039] (3) Cell culture flasks and 96-well culture plates;

[0040] (4) Test equipment: constant temperature water bath; carbon dioxide incubator; laminar flow ult...

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Abstract

The invention belongs to the technical field of disinfection, relates to a virus inactivation test, and particularly relates to a removal method of residual disinfectant in the virus inactivation test. The removal method comprises the steps of that (1) a preparation step is performed, specifically, a virus suspension is prepared; (2) a removal step is performed, specifically, first, disinfectant is sucked into a test tube, disinfectant removal treatment is performed after water bath, then ultracentrifugation is performed after 30-50 times dilution, then supernatant is removed, finally a cell maintenance solution is added for dilution and mixing again,, and the residual disinfectant is removed; and (3) a detection step is performed, specifically, virus suspension is sucked and added and mixed well, and a final sample is sucked for virus titer determination. According to the removal method of the residual disinfectant, the test time for disinfectant removal in the whole process can be shortened, the test efficiency of the virus inactivation test can be greatly improved, and the effects of disinfectants and neutralizers on cells and viruses in chemical neutralization methods are avoided.

Description

technical field [0001] The invention belongs to the technical field of disinfection and relates to a virus inactivation test, in particular to a method for removing residual disinfectant in the virus inactivation test. Background technique [0002] With the development of society and economy, changes in the environment, and changes in people's lifestyles, some new and unknown infectious diseases (AIDS, SARS, bird flu, Zika, hand-foot-mouth, Ebola, Echo virus, etc.), Hospital infection outbreaks and public health emergencies occur frequently, seriously affecting people's health and social stability. Infectious diseases caused by viruses are very easy to cause serious consequences due to their rapid outbreak and strong infectivity. Once it occurs, its transmission route and infection mechanism are difficult to be clarified in a short period of time, and targeted prevention and control measures are difficult to carry out in a timely manner , and the research and production of ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70
CPCC12Q1/70
Inventor 张磊钟昱文纪洵敏彭拓华张贤昌张吉凯郑小凌
Owner GUANGDONG PROVINCIAL INST OF BIOLOGICAL PROD & MATERIA MEDICA
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