Multiplex PCR rapid identification method for salmonella and listeria monocytogenes

A technology for Listeria and mononuclear cells, applied in biochemical equipment and methods, microbiological determination/inspection, etc., can solve the problems of cumbersome inspection methods, low specificity, time-consuming and labor-intensive, etc., and achieve identification and serological typing tests method fast effect

Inactive Publication Date: 2005-04-06
江苏瑞邦生物科技有限公司
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Problems solved by technology

Traditional methods of isolation, culture and serology are commonly used in the detection of Salmonella and Listeria monocytogenes. Different testing procedures and methods are used, and they are reported separately. It takes at least 5 to 7 days for

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Embodiment Construction

[0011] 1. Primers:

[0012] Two pairs of specific primers (primer I, primer II; hly-P 1 , hly-P 2 ), the above two pairs of specific primers were synthesized by Shanghai Boya Biological Co., Ltd. The size of the target fragment expected to be amplified is divided into Salmonella: 495bp; Listeria monocytogenes: 850bp; indicator band (Escherichia coli): 300bp.

[0013] salmonella:

[0014] Primer I: 5'-[ACT GGC GTT ATC CCT TTC TCT GCT G]-3'

[0015] Primer II: 5'-[ATG TTG TCC TGC CCC TGG TAA GAG A]-3'

[0016] Listeria monocytogenes:

[0017] hly-P 1 : 5'-[CCT AAG ACG CCA ATC GAA AAG AAA]-3'

[0018] hly-P 2 : 5’-[TAG TTC TAC ATC AAC TGA GAC AGA]-3’

[0019] Escherichia coli (indicator bacteria):

[0020] Primer I: 5'-[ACT GGC GTT ATC CCT TTC TCT GCT G]-3'

[0021] Primer II: 5'-[ATG TTG TCC TGC CCC TGG TAA GAG A]-3'

[0022] 2. Multiplex PCR:

[0023] 1. DNA template preparation:

[0024] 1) Salmonella DNA template preparation:

[0025] Inoculate Salmonella in nu...

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Abstract

The invention discloses a kind of appraisal bacterium, specially appraises the monk fungus (Salmonella), produces the uninuclear cell Liszt fungus (Listeria monocytogenes) the examination method. First (YAU_DSL1) separately uses the monk fungus and the backwoods coli the thermal crack solution to withdraw respective DNA template, produces the uninuclear cell Liszt fungus to use the enzyme solution to withdraw the DNA template, then carries on disposable PCR to expand increases the specific many genes, finally passes through the electricity to swim the appraisal. The invention through sample DNA, disposable at the same time to the monk fungus histidine transportation operon gene which possibly has, produces the uninuclear cell hemolysin gene to expand increases (YZU_DSL1) the material particle carries on with the certain existence backwoods coli expands increases, forms the massive genes duplication fragment, quite is convenient, can appraise whether for the monk fungus, produces the uninuclear cell Liszt fungus, and makes the judgement to the negative result reacting system accuracy. The invention more classical method bacterium separation, the appraisal and the blood serum school grades testing method fast, is accurate.

Description

technical field [0001] The invention discloses a detection method for identifying bacteria, in particular for identifying Salmonella and Listeria monocytogenes. Background technique [0002] Salmonella and Listeria monocytogenes are important zoonotic pathogens, and are the two most important foodborne pathogenic bacteria listed by the World Health Organization (WHO). Salmonella can not only cause animal diseases, but also cause typhoid fever, paratyphoid fever, sepsis, gastroenteritis and food poisoning in humans. In food poisoning around the world, the poisoning cases caused by Salmonella account for the first or second place; Listeria is A food-borne pathogen that can cause diseases such as abortion, sepsis and meningitis in animals and humans. Newborns, the elderly, pregnant women and immunocompromised patients are high-risk groups for Listeria infection. In countries all over the world, including the United States and other developed countries, food contamination by Sa...

Claims

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Application Information

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IPC IPC(8): C12Q1/04C12Q1/68
Inventor 焦新安黄金林潘志明文其乙刘秀梵周晓辉殷月兰孙林
Owner 江苏瑞邦生物科技有限公司
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