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High-efficiency method for screening salmonella and shigella in enterobacteriaceae

A technology for Salmonella and Shigella, which is applied in the direction of microorganism-based methods, biochemical equipment and methods, and microorganism measurement/inspection. It can solve problems such as high cost, complicated experimental methods, and time-consuming biochemical tests, and achieve low cost. , The experimental method is simple, the effect of less substrate

Inactive Publication Date: 2013-09-25
董根荣
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The current experimental methods for screening Salmonella and Shigella are relatively complex and require many substrates, and the screening of suspicious colonies is not thorough enough
However, biochemical tests for the purpose of identification in clinical laboratories are usually time-consuming and costly. Therefore, there is an urgent need for a method to quickly screen out Salmonella and Shigella from non-pathogenic bacteria growing on selective intestinal culture media. experiment method

Method used

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  • High-efficiency method for screening salmonella and shigella in enterobacteriaceae

Examples

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Embodiment 1

[0016] Example 1 of the present invention: Streak the specimen on a MacConkey agar plate, incubate at 37°C for 18 hours, then check the colorless colony growing on the plate; pick out a colorless colony, and use a part of it to detect oxidase activity; another part was inoculated on filter paper strips soaked with tryptophan, pyroglutamyl-4-methoxynaphthylamine and 4-methylumbelliferyl ioxynil caprylate and wetted with a drop of water or buffer . The appropriate amount of soaking liquid for filter paper is: 0.2ml of each of tryptophan, pyroglutamyl-4 methoxynaphthylamine and 4-methylumbelliferone ioxynil octanoate is soaked on 1 square inch of filter paper. 100 micrograms of 20 mM Tris buffer (pH 7.5), the filter paper used was Whatman No. 3 (Whatman Ltd. Maidstone England). The wetted inoculated strips were placed in a moisturizing vessel and incubated at 37°C for 2 hours, and then the filter paper strips were placed under a Wood lamp to detect fluorescence. Positive fluores...

Embodiment 2

[0017] Example 2 of the present invention: Streak the specimen on a MacConkey agar plate, incubate at 36°C for 20 hours, then check the colorless colony growing on the plate; pick out a colorless colony, and use a part of it to detect oxidase activity; another part was inoculated on filter paper strips soaked with tryptophan, pyroglutamyl-4-methoxynaphthylamine and 4-methylumbelliferyl ioxynil caprylate and wetted with a drop of water or buffer . The appropriate amount of soaking liquid for filter paper is: 0.3ml of each of tryptophan, pyroglutamyl-4 methoxynaphthylamine and 4-methylumbelliferone ioxynil octanoate is soaked on 1 square inch of filter paper. 160 μg of 20 mM Tris buffer (pH 7.5), the filter paper used is Whatman No. 3. The wet inoculated strips were placed in a moisturizing container and incubated at 36°C for 3 hours, and then the filter paper strips were placed under a Wood lamp to detect fluorescence. Positive fluorescence indicates the presence of C-8 estera...

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Abstract

The invention discloses a high-efficiency method for screening salmonella and shigella in enterobacteriaceae, and the method comprises steps of: spreading a specimen on a Mac Conkey agar plate, carrying out incubation at a temperature of 37 DEG C, and selecting a colorless bacterial colony, wherein a portion of the bacterial colony is used for detecting an oxidase activity; inoculating the other portion of the bacterial colony on a filter paper strip soaked with tryptophan, pyroglutamyl-4-methoxy naphthylamines and 4-methylumbelliferone ioxynil octoate, and moistening the inoculated filter paper strip with a drop of water or a drop of buffer; putting the moistened and inoculated filter paper strip in a moisture-maintaining vessel to subject to incubation, and putting the filter paper strip under a Wood lamp to detect fluorescence, wherein fluorescence-positive indicates that a C-8 esterase activity exists; and dropping a drop of a solution containing a diazo dye and iron chloride on the filter paper strip, so that if the filter paper strip shows fluorescence and invariant color, the bacterial colony on the filter paper strip can be identified as salmonella; and if the filter paper strip shows no fluorescence but invariant color, the bacterial colony on the filter paper strip can be identified as shigella. The method of the invention can detect activities of three kinds of enzymes at the same time by employing a method of mixing substrates, the experiment method is simple, a dosage of substrates is lesser, cost is low, and salmonella and shigella can be screened out from nonpathogenic bacteria in enterobacteriaceae efficiently.

Description

technical field [0001] The invention relates to a rapid screening method for Salmonella and Shigella in Enterobacteriaceae. Background technique [0002] Enterobacteriaceae is composed of a group of Gram-negative bacilli with similar biological characteristics, facultative anaerobic, oxidase-negative, some of which have a strong pathogenic effect on humans, including intestinal infection, sepsis and urinary tract infection many infectious diseases. Salmonella and Shigella are very important pathogenic bacteria, which can cause diarrhea and gastroenteritis. The current experimental methods for screening Salmonella and Shigella are relatively complex and require many substrates, and the screening of suspicious colonies is not thorough enough. However, biochemical tests for the purpose of identification in clinical laboratories are usually time-consuming and costly. Therefore, there is an urgent need for a method to quickly screen out Salmonella and Shigella from non-pathogen...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/04C12Q1/34C12Q1/37C12Q1/44C12R1/42C12R1/01
Inventor 董根荣
Owner 董根荣
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