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Method for rapidly determining titer of bacteriophage of Enterobacter sakazakii

A technology of Enterobacter sakazakii and bacteriophage is applied in the field of rapid determination of phage titer of Enterobacter sakazakii, which can solve the problems of inability to obtain plaques, inability to obtain titers, and many operation steps.

Inactive Publication Date: 2010-07-14
CHINESE ACAD OF INSPECTION & QUARANTINE
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  • Abstract
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Problems solved by technology

This method has a lot of steps, and when the Enterobacter sakazakii phage titer is determined, it is impossible to obtain obvious phage plaques in the area, so that an accurate titer cannot be obtained

Method used

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  • Method for rapidly determining titer of bacteriophage of Enterobacter sakazakii

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Embodiment

[0046] This example provides a typical example of the method for rapidly determining the phage titer of Enterobacter sakazakii of the present invention.

[0047] The method includes the following steps:

[0048] 1. Preparation of phage:

[0049] Preparation of the bacterial suspension of Enterobacter sakazakii: Inoculate Enterobacter sakazakii (from the Food Safety Microorganism Culture Preservation Management Center of the Chinese Academy of Inspection and Quarantine) into 10 mL of nutrient broth culture solution (the nutrient broth was purchased from Beijing Land Bridge Technology Co., Ltd. product, China, prepared in nutrient broth (culture solution) according to the product instructions), and cultured at 36°C±1°C for 8 to 12 hours. Determination of OD by spectrophotometer 600 , OD 600 If it is greater than 0.5, it indicates that Enterobacter sakazakii is in the logarithmic growth phase, and the culture is stopped. At this time, the sensitivity of Enterobacter sakazaki...

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Abstract

The invention relates to a method for rapidly determining the titer of bacteriophage of Enterobacter sakazakii, comprising the following steps of: A. preparing a bacteriophage concentration gradient suspension; B. preparing an Enterobacter sakazakii suspension; C. sequentially adding an Enterobacter sakazakii culture solution, the Enterobacter sakazakii suspension in the step B and the acteriophage concentration gradient suspension in the step A in a micropore plate; and D. scanning and determining in real time, scanning and determining a growth curve in a dynamics determining mode in real time under a certain culture temperature and wave length by utilizing a microplate reader. By using the determining method, the titer of the bacteriophage of the Enterobacter sakazakii can be simply, rapidly and efficiently determined.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a rapid determination method for enterobacter sakazakii phage titer. Background technique [0002] Enterobacter sakazakii is a gram-negative bacillus with perinatal flagella, motility, no spores, facultative anaerobic, which can cause meningitis, enterocolitis, bacteremia, etc., especially for infants and young children and immunity The low population has a strong pathogenicity, and the mortality rate is as high as 50%-75%. Enterobacter sakazakii exists widely in the natural environment, and because of its advantages such as wide growth temperature range, high temperature resistance, drying resistance, and bacterial film protection layer, once the bacteria pollutes the food production environment, it is easy to cause damage to the entire production process. pollution and is not easy to eradicate. At present, physical and chemical methods such as disinfectant, high temperature or ultr...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12R1/92
Inventor 袁飞
Owner CHINESE ACAD OF INSPECTION & QUARANTINE
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