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Primers special for detecting coxiella burnetii

A technology of Coxiella berghei and primer sets, applied in the biological field, can solve the problems of taking a long time and difficult to meet the rapid diagnosis of outbreaks, and achieve the effects of high sensitivity, high specificity, and convenient and simple identification.

Inactive Publication Date: 2013-02-13
MICROBE EPIDEMIC DISEASE INST OF PLA MILITARY MEDICAL ACAD OF SCI
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  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

Traditional ordinary PCR and real-time fluorescent quantitative PCR methods often take a long time or require specific instruments, which are difficult to meet the needs of rapid diagnosis of outbreaks

Method used

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  • Primers special for detecting coxiella burnetii
  • Primers special for detecting coxiella burnetii
  • Primers special for detecting coxiella burnetii

Examples

Experimental program
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Effect test

Embodiment 1

[0036] Embodiment 1, the design and preparation of special primers for detection of Coxiella basilica

[0037] The sequences of specific primers designed according to the whole genome sequence of Coxiella basilica Xinqiao strain are as follows:

[0038] F3 (sequence 1): 5'-TTCAATAAGCGTTTTATCGCT-3';

[0039] B3 (sequence 2): 5'-ACAACCATTTAAACCGACAAG-3';

[0040] FIP (sequence 3): 5'-AAGGCTTCCATCAGTGTGAATTATTTGAAGTTATCGAAAGATTGCGT-3';

[0041] BIP (sequence 4): 5'-CGAATGGTGTCGCGTGTTTCCCCCTTTATCTCATTTCCGG-3';

Embodiment 2

[0043] Example 2, the application of special primers for detection of Coxiella bainiella

[0044] 1. Preparation of samples of Coxiella basilica

[0045] The yolk sac membrane of chicken embryos infected with Coxiella beinii Xinqiao strain was added to broth for grinding to prepare the yolk sac membrane grinding liquid of chicken embryos infected with Coxiella beinii. Ditrizoate density gradient centrifugation was used for purification, and the purified Coxiella basidioides was extracted with DNeasy Blood&Tissue Kit. The specific extraction method was found in the kit manual. Finally, Nanodrop1000 was used to measure the DNA concentration of the extracted cells. Concentration (ng / μL). The genome size of Coxiella basilica Xinqiao strain is 2Mbp, according to the concentration of bacterial DNA (ng / μL) and the size of the genome (2Mbp), calculate the mole of purified Coxiella basilica Xinqiao strain Count, and finally calculate the concentration of the purified Coxiella basilar...

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Abstract

The invention discloses primers special for detecting coxiella burnetii and provides a loop mediated isothermal amplification primer group for detecting the coxiella burnetii. The loop mediated isothermal amplification primer group comprises a primer 1, a primer 2, a primer 3 and a primer 4. Nucleotide sequences of the primer 1, the primer 2, the primer 3 and the primer 4 are respectively a sequence 1, a sequence 2, a sequence 3 and a sequence 4 in a sequence table in sequence. The primers special for detecting the coxiella burnetii have the advantages of being capable of performing amplification reaction only at constant temperature, free of special devices, high in specificity, rapid, efficient, capable of finishing the amplification reaction within 60 minutes, high in sensitivity, convenient and simple to identify and particularly suitable for rapid detection of clinical specimens.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a special primer for detecting Coxiella bezieri. Background technique [0002] Coxiella beinii is a rickettsia-like pathogen isolated from an Australian patient for the first time in 1935. It is an obligate intracellular parasitic, Gram-negative acidophilic bacterium. The disease caused by Coxiella basilica is called Q fever (Q fever). Q fever is divided into two clinical types, acute and chronic. Acute Q fever manifests as acute flu-like symptoms, mainly characterized by headache, fever, and pneumonia, while chronic Q fever mainly manifests as long-term fever, often accompanied by endocarditis, hepatitis, and osteomyelitis. The infectious dose of Coxiella beinii is extremely low, and a single pathogen can cause disease. Therefore, this pathogen has been used as a biological weapon by some countries, and may also be used as a bioterrorist agent by terrorist organizations. The repo...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11C12R1/01
Inventor 王锡乐温博海熊小路齐永段长松李佳明焦俊贾引军龚文平
Owner MICROBE EPIDEMIC DISEASE INST OF PLA MILITARY MEDICAL ACAD OF SCI
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