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Quantitative detection method of salmonella living body in water

A quantitative detection method and technology for Salmonella, applied in the determination/inspection of microorganisms, biochemical equipment and methods, resistance to vector-borne diseases, etc., can solve the problems of difficult quantitative detection, inconsistent types, complicated operations, etc. Feasible, improved specificity

Inactive Publication Date: 2011-08-17
XI'AN UNIVERSITY OF ARCHITECTURE AND TECHNOLOGY
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Problems solved by technology

Most of these methods have the problems of complex operation, susceptibility to interference, and difficulty in quantitative detection
There are many kinds of Salmonella, and their characters often change in the water environment, and the actual biochemical reaction results often do not match the types in the Salmonella identification table, resulting in detection failures
Nucleic acid detection method is to use PCR technology to detect the conserved gene fragments of Salmonella to achieve the purpose of accurate detection. However, its biggest drawback is that it cannot distinguish between living and dead bacteria, and it is impossible to in-depth study of Salmonella in water environment by simply using PCR technology. human health impact
Therefore, there is currently no quantitative method for the detection of Salmonella living organisms in water.

Method used

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  • Quantitative detection method of salmonella living body in water
  • Quantitative detection method of salmonella living body in water
  • Quantitative detection method of salmonella living body in water

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Experimental program
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Effect test

Embodiment Construction

[0029] 1. Equipment and reagents

[0030] (1) PCR instrument;

[0031] (2) Nucleic acid horizontal electrophoresis instrument;

[0032] (3) Gel imager;

[0033] (4) Constant temperature incubator

[0034] (5) High pressure steam sterilizer

[0035] (6) Stainless steel vacuum filter

[0036] (7) microwave oven

[0037] (8) 0.22μm mixed cellulose ester microporous membrane

[0038] (9) Salmonella typhimurium;

[0039] (10) Xylose lysine deoxycholate agar medium (XLD)

[0040] (11) Bismuth sulfite agar medium (BS)

[0041] (12) Magnesium Chloride Malachite Green Enrichment Solution

[0042] (13) Peptone

[0043] (14) Agarose

[0044] (15) Tris dry powder

[0045] (16) Na 2 EDTA·2H 2 o

[0046] (17) Nucleic acid dyes

[0047] (18) DNA markers

[0048] (19) 6×Loading buffer

[0049] (20) Taq enzyme

[0050] (21) dNTPs

[0051] (22) PCR buffer

[0052] (23) General primers for Salmonella, including upstream primer 139 and downstream primer 141; the upstream and ...

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Abstract

The invention discloses a quantitative detection method of salmonella living body in water, which adopts a multiple-tube fermentation method to preliminarily screen living bacteria taking salmonella as the principal component from water sample according to the growth characteristics of the salmonella and the conservative property of invA gene. By utilizing universal primers 139 and 141 of the salmonella, the quantitative detection method can be used for distinguishing the positive colony of the salmonella by polymerase chain reaction (PCR) amplification. The quantitative detection method can be used for detecting the salmonella living body in water in a quantitative way by a most probable number (MPN) counting method. The method can accurately detect the content of the salmonella living body in water, is high in specificity as well as simple and convenient to operate, and is suitable for detecting multiple water samples such as surface water, sewage and the like.

Description

technical field [0001] The invention relates to a detection method for surface water and sewage, in particular to a quantitative detection method for Salmonella living organisms in water. The method combines multi-tube fermentation technology, PCR technology and the most probable number (MPN) counting method to enable accurate quantitative detection. Live Salmonella levels in water. It is suitable for the quantitative detection of live Salmonella in surface water, sewage and other water samples. Background technique [0002] The current detection of Salmonella can be divided into three categories: culture method, immunological detection method and nucleic acid detection method. The culture method and immunological detection method are usually based on the growth characteristics of Salmonella, using a selective medium to make Salmonella a dominant bacterial group, and then based on the biochemical characteristics of Salmonella and the surface antigen characteristics of the b...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/10C12Q1/06
CPCY02A50/30
Inventor 张崇淼王晓昌船水尚行
Owner XI'AN UNIVERSITY OF ARCHITECTURE AND TECHNOLOGY
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