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Multiplex PCR based primer pair and kit for detecting multiple intestinal pathogens

A technology for detecting primers and pathogenic bacteria, which is applied in the field of PCR applications, and can solve the problems that the detection and identification of various pathogenic bacteria cannot be achieved.

Active Publication Date: 2015-04-22
AGCU SCIENTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the methods of biochemical identification, quantitative PCR and ELISA are used to identify a small number of bacteria, which cannot achieve parallel detection and identification of multiple pathogenic bacteria in a short period of time.

Method used

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  • Multiplex PCR based primer pair and kit for detecting multiple intestinal pathogens
  • Multiplex PCR based primer pair and kit for detecting multiple intestinal pathogens
  • Multiplex PCR based primer pair and kit for detecting multiple intestinal pathogens

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0139] Example 1 Bacteria detection test

[0140] The DNA sample of Vibrio cholerae O1, O139 group, non-O1 / O139 group used in the present embodiment, the DNA sample of Listeria monocytogenes, the DNA sample of pathogenic Escherichia coli (EAEC, EPEC, ETEC, EIEC, EHEC) DNA samples, DNA samples of Shigella dysenteriae, DNA samples of EV71, DNA samples of enterohemorrhagic E. As a gift from Zhejiang CDC, 5 samples of each kind were obtained. These samples were cultured and identified in advance, and the relevant toxins were identified (see Table 6 for the sample number and the results of the culture identification).

[0141] Table 6 Sample number and culture identification results

[0142]

[0143]

[0144]

[0145] The primer set described in Table 2 was used for multiplex amplification, the reaction system was prepared according to Table 3, the above-mentioned extracted DNA template was added respectively, and the reaction was carried out according to the reaction pro...

Embodiment 3

[0152] Embodiment 3 specificity test

[0153] The following samples were used in this specificity test: Clostridium difficile (non-toxigenic) DNA sample, Campylobacter jejuni DNA sample, Yersinia enterocolitica DNA sample, Staphylococcus aureus DNA sample, meat The DNA samples of Clostridium toxin and Bacillus cereus were donated by Zhejiang CDC, with 5 samples of each type, which were cultured and identified in advance. The sample list is as follows:

[0154] Table 7 Specific test samples

[0155] sample number bacteria J1 Clostridium difficile (non-toxigenic) J2 Campylobacter jejuni J3 Yersinia enterocolitica J4 Staphylococcus aureus J5 Clostridium botulinum J6 Bacillus cereus

[0156] The primer set described in Table 2 was used for multiplex amplification, the reaction system was prepared according to Table 3, the above-mentioned extracted DNA template was added respectively, and the reaction was carried out according t...

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Abstract

The invention relates to a multiplex PCR based primer pair and kit for detecting intestinal pathogens, particularly relates to a multiplex PCR based primer pair and kit for detecting 14 intestinal pathogens, and belongs to the technical field of PCR application. The 14 intestinal pathogens comprise vibrio cholerae (group O1 phage, group O139 phage and group non-O1 / O139 phage), listeria monocytogenes, enteropathogenic escherichia coli (EPEC), enterohemorrhagic escherichia coli (EHEC), enterotoxigenic escherichia coli (ETEC), enteroinvasive escherichia coli (EIEC), enteroaggregative escherichia coli (EAEC), shigella, intestinal virus EV71, enterohemorrhagic escherichia coli O157:H7, clostridium difficile, vibrio parahaemolyticus, salmonella enteritidis and salmonella typhimurium. The multiplex PCR based primer pair and kit can change the situation that only a few intestinal pathogens can be detected in one time and can be used for detecting 14 intestinal pathogens and 26 genes simultaneously.

Description

technical field [0001] The invention relates to a multiple PCR detection primer set and kit for intestinal pathogenic bacteria, and belongs to the technical field of PCR application. More specifically, it involves the detection of 14 kinds of intestinal pathogenic bacteria, including: Vibrio cholerae (O1 group, O139 group, non-O1 / O139 group), Listeria monocytogenes, enteropathogenic Escherichia coli (EPEC), intestinal Hemorrhagic Escherichia coli (EHEC), Enterotoxigenic Escherichia coli (ETEC), Enteroinvasive Escherichia coli (EIEC), Enteroaggregative Escherichia coli (EAEC), Shigella, Enterovirus EV71, Enterohemorrhagic Escherichia coli O157 : H7, Clostridium difficile, Vibrio parahaemolyticus, Salmonella enteritidis, Salmonella typhimurium. Background technique [0002] Infectious diseases caused by intestinal pathogenic bacteria are the main factors that endanger public health security. Most countries have taken corresponding measures to detect enteric pathogenic bacter...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68C12Q1/04C12N15/11C12R1/93C12R1/01C12R1/19C12R1/63
CPCC12Q1/689C12Q1/701C12Q2600/16Y02A50/30
Inventor 郑卫国周丽萍葛斌文葛海鹏郭育林
Owner AGCU SCIENTECH
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