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Protein combination for diagnosing Q fever

A composition and protein technology, applied in the field of protein composition for diagnosing Q fever, can solve the problems of difficult promotion and use, high cost, complicated process and the like

Inactive Publication Date: 2013-07-31
MICROBE EPIDEMIC DISEASE INST OF PLA MILITARY MEDICAL ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these detection methods require a large number of samples, the steps are cumbersome and time-consuming, and all of them require purified whole bacteria of Coxiella beinii as antigens, while the protection requirements for extracting and purifying Coxiella beinii are high, the process is complicated, and the cost is expensive , so these methods are difficult to be widely promoted and used

Method used

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  • Protein combination for diagnosing Q fever
  • Protein combination for diagnosing Q fever
  • Protein combination for diagnosing Q fever

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0082] The preparation of embodiment 1, 7 kinds of antigenic proteins

[0083] The vector pET-32a(+) was purchased from Novagen, the product catalog number is 69015. pQE30 was purchased from Qiagen Company, the product catalog number is 33203.

[0084] Escherichia coli BL21 was purchased from Novagen, the product catalog number is 69450. Escherichia coli M15 was purchased from Novagen, the product catalog number is 34210.

[0085] 1. Protein sequences and coding gene sequences of seven antigenic proteins

[0086] The protein sequences and coding gene sequences of the seven antigenic proteins are shown in Table 1.

[0087] Table 1, protein and gene sequence

[0088] protein name

protein sequence

coding gene sequence

OMP

SEQ ID NO: 1

SEQ ID NO: 8

Com1

SEQ ID NO: 2

SEQ ID NO: 9

wxya

SEQ ID NO: 3

SEQ ID NO: 10

Mip

SEQ ID NO: 4

SEQ ID NO: 11

DnaK

SEQ ID NO: 5

SEQ ID NO: 12

GroEL ...

Embodiment 2

[0115] Embodiment 2, protein chip and its preparation

[0116] Mouse IgG was purchased from Beijing Hongyue Innovation Biotechnology Co., Ltd., the product catalog number is M401-M422. Human IgG was purchased from Beijing Hongyue Innovation Biotechnology Co., Ltd., the product catalog number is M401-M422.

[0117] 1. Protein chip for detecting human serum and its preparation

[0118] The structure of the protein chip: it is composed of an aldylated slide substrate, a detection point, a positive quality control point and a negative quality control point, and the detection point, the positive quality control point and the negative quality control point are all spotted on the substrate. There were 5 replicates for each detection point, positive quality control point and negative quality control point. Detection points, positive quality control points and negative quality control points are arranged in a matrix on the substrate.

[0119] The detection points are divided into se...

Embodiment 3

[0128] Embodiment 3, diagnosis Q fever

[0129] Cy5-labeled goat anti-mouse IgG was purchased from KPL, USA, and the product catalog number was 072-02-18-06.

[0130] Cy5-labeled goat anti-human IgG was purchased from KPL, USA, and the product catalog number was 072-02-10-06.

[0131] Each 1 liter of phosphate buffered saline (PBS) is prepared as follows: 8gNaCl, 0.2gKCl, 3.53gNa 2 HPO 4 .12H 2 O, 0.24g KH 2 PO 4 Mix with water, and dilute to 1 liter with water, and the pH value of the obtained solution is 7.4, that is, the PBS buffer solution is obtained.

[0132] Every 1 liter of PBST buffer was prepared as follows: phosphate buffer (PBS) was mixed with Tween-20, and the ratio of phosphate buffer (PBS) and Tween-20 was 1 liter: 1 ml to obtain PBST buffer.

[0133] The mice were 6-week-old male BALB / c mice purchased from the Experimental Animal Center of the Academy of Military Medical Sciences.

[0134] 1. Diagnosis of Q fever in mice

[0135] (1) The composition of ...

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Abstract

The invention discloses a protein combination for diagnosing a Q fever. The protein combination consists of the following proteins from (1) to (7): (1) a protein of which the amino acid sequence is represented by SEQ ID NO: 1, (2) a protein of which the amino acid sequence is represented by SEQ ID NO: 2, (3) a protein of which the amino acid sequence is represented by SEQ ID NO: 3, (4) a protein of which the amino acid sequence is represented by SEQ ID NO: 4, (5) a protein of which the amino acid sequence is represented by SEQ ID NO: 5, (6) a protein of which the amino acid sequence is represented by SEQ ID NO: 6, and (7) a protein of which the amino acid sequence is represented by SEQ ID NO: 7. The protein combination and a protein chip can perform quick and accurate serology analysis and diagnosis on blood serum of a human or an animal having the Q fever; the result is accurate and reliable; moreover, the operation is simple, the number of used samples is low, the time and the labor are saved, and the protein combination is desired to be substituted for the conventional complicated, tiring and time-consumed serology diagnosis method for Q fever serology diagnosis.

Description

technical field [0001] The invention relates to a protein composition for diagnosing Q fever. Background technique [0002] Coxiella burnetii is a small Gram-negative bacillus parasitic in obligate phagocytic cells. The bacterium has strong resistance to the external environment and has strong infectivity to humans and animals. It can enter the body through aerosol diffusion through the respiratory tract, causing infection in humans and animals and Q fever. [0003] Q fever is a zoonotic disease with worldwide distribution and is widely distributed in my country. Human Q fever is divided into two types: acute and chronic. Acute Q fever is characterized by fever, headache, and muscle aches as the main symptoms, often accompanied by pneumonia. Some patients may also develop hepatitis, myocarditis, and even meningitis. If acute Q fever is not treated in time or completely, it can become chronic, causing chronic hepatitis, endocarditis, osteomyelitis, etc. [0004] At present...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/29
Inventor 熊小路温博海王锡乐
Owner MICROBE EPIDEMIC DISEASE INST OF PLA MILITARY MEDICAL ACAD OF SCI
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