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Multi-PCR detecting method capable of detecting mononuclear hyperplasia, staphylococcus aureus and enterococcus faecium simultaneously

A technology for Staphylococcus and Enterococcus faecium, applied in the field of multiple PCR detection, can solve the problems of multiple PCR detection methods that have not been reported yet, complex pathogen types, cumbersome steps, etc., and achieve the effect of simple method, high sensitivity and low cost

Active Publication Date: 2017-05-10
INNER MONGOLIA AUTONOMOUS REGION ACAD OF AGRI & ANIMAL HUSBANDRY SCI
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AI Technical Summary

Problems solved by technology

So far, the veterinary clinical diagnosis of the disease mainly relies on the isolation and identification of the pathogen. However, due to the complexity of the pathogenic species, especially the large number of bacteria of the genus Listeria, the identification has the disadvantages of cumbersome steps and long time. The isolation and test of Listeria karyocytogenes have high risks. Therefore, the establishment of a rapid and specific PCR diagnostic method has important clinical value for the early diagnosis and treatment of the disease
[0006] At present, the multiple PCR detection methods established for these three bacteria have not been reported at home and abroad.

Method used

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  • Multi-PCR detecting method capable of detecting mononuclear hyperplasia, staphylococcus aureus and enterococcus faecium simultaneously
  • Multi-PCR detecting method capable of detecting mononuclear hyperplasia, staphylococcus aureus and enterococcus faecium simultaneously
  • Multi-PCR detecting method capable of detecting mononuclear hyperplasia, staphylococcus aureus and enterococcus faecium simultaneously

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Embodiment 1

[0029] Example 1. Multiplex PCR method

[0030] 1. Test strain

[0031] Clinical isolates: Listeria monocytogenes, Listeria ovis, Staphylococcus aureus, Enterococcus faecium, Coccus faecalis, Enterococcus mantesii, Streptococcus, Pasteurella, Escherichia coli, the strains used in the test were all Preserved by the Institute of Veterinary Medicine, Inner Mongolia Academy of Agriculture and Animal Husbandry Sciences.

[0032] 2. Main instruments and reagents

[0033] The Biophotometer trace nucleic acid detector was purchased from Effendorf. DL 600 DNA Marker and bacterial genomic DNA extraction kit were purchased from Tiangen Biochemical Technology (Beijing) Co., Ltd. PremixTaq was purchased from Beijing Quanshijin Biotechnology Co., Ltd. Primers were synthesized by Sangon Bioengineering (Shanghai) Co., Ltd.

[0034] 3. Primer design

[0035] According to the heat-resistant hyaluronidase NUC of Staphylococcus aureus, the genus-specific primers NUC-1 and NUC-2 were designe...

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Abstract

The invention discloses a multi-PCR detecting method capable of detecting mononuclear hyperplasia, staphylococcus aureus and enterococcus faecium simultaneously. The multi-PCR detecting method capable of simultaneously detecting three pathogenic bacteria, namely, the mononuclear hyperplasia, the staphylococcus aureus and the enterococcus faecium, is established by designing premiers respectively according to the hlyA gene of the mononuclear hyperplasia, the NUC gene of the staphylococcus aureus and the ddl gene of the enterococcus faecium. According to the method, the DNA fragments of the mononuclear hyperplasia, the staphylococcus aureus and the enterococcus faecium can be amplified specifically; the lowest detecting amounts are 1.07 pg, 11.6 pg, and 1.21 pg respectively. The method does not have a cross reaction to other common pathogenic bacteria. The multi-PCR reaction established by the method has the characteristics of high specificity and high sensitivity; a convenient, quick and accurate method is provided for quick detection, authentication and epidemiological investigation of the mononuclear hyperplasia, the staphylococcus aureus and the enterococcus faecium in clinical work.

Description

technical field [0001] The invention relates to a multiple PCR detection method, in particular to a multiple PCR detection method with short time consumption, high efficiency and high sensitivity, which can be directly applied to simultaneous detection of Listeria monocytogenes, Staphylococcus aureus and Enterococcus faecium method. Background technique [0002] Listeria monocytogenes is a zoonotic disease that can cause skin infections, gastrointestinal disturbances, and sepsis. Listeria monocytogenes is mainly infected through the intestinal tract, and the first target organ invaded from the intestinal tract is the liver. Listeria monocytogenes can reproduce in large numbers in the liver, and it will not stop until the body's cellular immune response is enhanced. Listeria monocytogenes, similar to Mycobacterium tuberculosis and Toxoplasma gondii, can reduce the function of cellular immunity of animal mothers, thereby causing immunodeficiency in pregnant animal mothers, l...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/14C12Q1/04C12N15/11
CPCC12Q1/686C12Q1/689C12Q2600/16C12Q2537/143
Inventor 刘昆宋越刘威张月梅裴乐陈伟赵世华
Owner INNER MONGOLIA AUTONOMOUS REGION ACAD OF AGRI & ANIMAL HUSBANDRY SCI
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