A method for detecting formaldehyde inactivation effect of toxin-producing Pasteurella multocida toxin by using Vero cells

Abstract

The invention belongs to the field of biotechnology, and in particular relates to a method for using Vero cells to detect the formaldehyde inactivation effect of toxin-producing Pasteurella multocida toxin, comprising: Step 1: preparing a toxin-producing Pasteurella multocida bacterial liquid; Step 2: Inactivated bacteria liquid for standby; Step 3: Preparation of toxin-producing Pasteurella multocida toxin liquid; Step 4: After digesting and counting the cultured monolayer Vero cells, use MEM medium containing 2% fetal bovine serum To dilute the cell suspension, add 100 μl of MEM medium to each well of the multi-well experimental well plate; add the bacterial toxin solution to the well according to the method of equal dilution; add 100 μl of Vero cell suspension to each well, mix well and place it at 36~ Observe the results after seven days of cultivation in a CO2 incubator at 38°C; cells with stringy lesions are judged as positive, bacterial aggregations are judged as suspicious, and no lesions are judged as negative. The method can more accurately detect whether the toxin-producing Pasteurella multocida toxin is completely or maximally inactivated, thereby well ensuring the safety of related vaccines.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a method for detecting the formaldehyde inactivation effect of toxin-producing Pasteurella multocida toxin by using Vero cells. Background technique [0002] Toxigenic pasteurella multocida (T + Pm) is the main pathogen of porcine infectious atrophic rhinitis (Atrophicrhinitis, AR). Porcine atrophic rhinitis can cause turbinate atrophy in pigs, destroy the immune barrier of the nasal mucosa, and then cause other respiratory diseases, resulting in significant economic losses for the pig industry. Studies have shown that toxigenic Pasteurella multocida can cause progressive atrophic rhinitis, and the toxigenic Pasteurella multocida toxin is the main pathogenic factor. Pasteurella multocida is divided into 6 serotypes such as A, B, D, E, and F according to the capsule. It has been reported that T + Pm mainly produces D-type toxins, and also has A-type toxins, while T + Pm was mainly...

AI Technical Summary

Problems solved by technology

Due to the different culture time of strains, individual differences in guinea pigs and personnel operating errors, false negatives may be caused. At the same time, the skin necrosis test of guinea pigs uses inactivated bacteria liquid, and the proportion of bacteria is small, which cannot reasonably reflect the production of toxins in the bacteria. Whether Pasteurella toxin is inactivated

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