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Legionella pneumophilia antigen near-infrared fluorescent detection kit and purpose thereof

A technology for Legionella pneumophila and fluorescence detection is applied in the field of near-infrared fluorescence detection kits for Legionella pneumophila antigens, which can solve the problem that the detection technology level of Legionella pneumophila is lagging behind, the antibody content cannot reach the detection level, and it is not suitable for clinical detection. application and other issues, to achieve high sensitivity, good specificity, and good specificity

Inactive Publication Date: 2016-08-03
85 HOSPITAL OF PEOPLES LIBERATION ARMY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the RIA method is radioactive, EIA takes a long time, and the antibody content in the serum at the initial stage of infection cannot reach the detection level, so it is not suitable for clinical detection applications
At this stage in my country, the level of detection technology for Legionella pneumophila is still relatively lagging (Zhu Jiaxin, Chen Wensi, Huang Jing, et al. Preliminary evaluation of a domestic Legionella pneumophila antigen detection kit[J]. Journal of Practical Medicine, 2010,26 (9): 1637-1638)

Method used

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  • Legionella pneumophilia antigen near-infrared fluorescent detection kit and purpose thereof
  • Legionella pneumophilia antigen near-infrared fluorescent detection kit and purpose thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] 1. Sputum collection and storage

[0055] Under the guidance of medical staff, the patient coughs up the sputum in the deep part of the respiratory tract forcefully, spits it into a sterile jar, and immediately closes the lid tightly and saves it for later use. The samples to be processed should be stored in a refrigerator at 4°C.

[0056] 2. Urine collection and storage

[0057] Under the guidance of medical staff, the patient washes and collects the first urine in the morning, and the collected urine is stored in a clean and sterile container for future use. Urine samples can be stored at 2-8°C for 72 hours. If they need to be stored for a longer period of time, the samples must be stored at -20°C. Repeated freezing and thawing of samples should be avoided, otherwise erroneous experimental results will be produced. Samples should not be stored in an automatic defrost refrigerator.

[0058] 3. Antigen preparation:

[0059] Purchase Legionella pneumophila strains fr...

Embodiment 2

[0074] Comparison experiment of Legionella pneumophila antigen near-infrared fluorescence detection method and sputum bacterial culture:

[0075] Sputum bacterial culture:

[0076] The sputum was pretreated (the method of combining 1ug / ml trypsin solution with glass fragment shaking was used to pretreat the sputum before inoculation and culture), and the pretreated samples were immediately inoculated on the GVPC selective medium. Gram-stained colonies of suspected Legionella pneumophila were inoculated on BCYE-a and BCYE-Cys agar plates with negative Gram-stained colonies, and cultured in a 37°C incubator for 2 days. Any colony that grows on BCYE-a medium but does not grow on BCYE-Cys medium can be considered as Legionella pneumophila.

[0077] Kit detection:

[0078] The sample to be tested is a sputum dilution, which is diluted with 0.01M pH7.2 PBS buffer, and the dilution ratio is 1:1.

[0079] The sample to be tested and the kit are equilibrated to room temperature to s...

Embodiment 3

[0088] Cross experiment:

[0089] Add the prepared Enterococcus faecalis, Enterococcus faecium, Escherichia coli, Acinetobacter baumannii, Klebsiella, Lactobacillus, Neisseria gonorrhoeae, green Pseudomonas pyogenes, Escherichia coli, Mycoplasma hominis, Ureaplasma urealyticum, Staphylococcus aureus, Streptococcus and Helicobacter pylori (1×10 6 CFU / ml), to carry out the cross experiment detection to detect whether the above bacteria have an impact on the test results of the kit. The results of the cross experiment between the Legionella pneumophila antigen near-infrared fluorescence detection reagent and the bacteria are shown in Table 2. There was no cross experiment in each bacteria cross experiment. Cross-reactivity occurs:

[0090] Table 2

[0091] cross

[0092] As can be seen from Table 1 in Example 2 and Table 2 in Example 3, in the detection of a total of 30 clinical samples, 4 cases were detected as LP positive by near-infrared fluorescence detection meth...

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Abstract

The invention relates to the technical field of biology, and especially relates to a legionella pneumophilia antigen near-infrared fluorescent detection kit and a purpose thereof. The invention provides a legionella pneumophilia antigen near-infrared fluorescent detection kit, which comprises reagent strips on a backboard, ad the reagent strips are a sample pad, a glass cellulose membrane labeled with an immunofluorescence probe, a cellulose nitrate membrane coated with a LP antibody and absorbent paper. The provided detection kit takes patient urine or sputum as a detection sample, by combining with a near-infrared fluorescent detection technology, the specific and soluble LP antigen in the patient sample can be rapidly and accurately detected.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a near-infrared fluorescence detection kit for Legionella pneumophila antigen and its application. Background technique [0002] The model bacterium of Legionella is Legionella pneumophila (Legionella Pneumophila), accounts for 90% of Legionella (Lu Feng, Cheng Yibin, Wang Zhong, et al. Epidemiological investigation of Legionella pneumophila infection in patients with respiratory diseases[J] .Environmental and Health Impurities, 2012,27(3):203-205), its sporadic cases account for 1%-15% of community-acquired pneumonia, 3.8% of hospital-acquired pneumonia, and 4% of difficult-to-diagnose atypical pneumonia -11% (Lu Feng, Jin Yinlong, Cheng Yibin, et al. Epidemiological situation of Legionnaires' disease [J]. Foreign Medicine: Hygiene, 2008, 35(2): 78). Legionnaires disease is a bacterial respiratory infectious disease mainly caused by Legionella pneumophila. The detection methods fo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N33/543G01N21/64C07K16/12
Inventor 姜小华
Owner 85 HOSPITAL OF PEOPLES LIBERATION ARMY
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