Reagent for auxiliary diagnosis of tubercular meningitis

A technology for tuberculous meningitis and pulmonary tuberculosis, applied in the field of medical diagnosis, can solve the problems of difficult diagnosis, diagnosis lag, and long time, and achieve the effects of shortening the diagnosis time, improving accuracy, and reducing the incidence rate

Inactive Publication Date: 2021-08-03
BEIJING CHEST HOSPITAL CAPITAL MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The clinical symptoms of tuberculous meningitis are diverse, and the diagnosis requires cerebrospinal fluid smear, culture or positive Xpert (Suzaan Marais, Guy Thwaites, Johan FSchoeman. et al. Tuberculous meningitis: a uniform case definition for use clinical research. Lancet Infect Dis 2010; 10 :803–12), and most tuberculous meningitis are secondary to pulmonary tuberculosis, and doctors pay attention only after neurological symptoms appear. Therefore, the diagnosis still has a certain lag, and it is difficult to diagnose clinically. After a long time, there are many sequelae, which seriously affect the quality of life of patients

Method used

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  • Reagent for auxiliary diagnosis of tubercular meningitis
  • Reagent for auxiliary diagnosis of tubercular meningitis
  • Reagent for auxiliary diagnosis of tubercular meningitis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1: Different groups of people in other groups of feces

[0024] 1, tuberculosis determine the diagnostic criteria:

[0025] Swipet-positive tuberculosis: one of the following items:

[0026] (1) Two specimens sputum smear Antibacterial positive.

[0027] (2) 1 sputum specimens antibiotics and positive tuberculosis

[0028] (3) 1 part of the sputum smear anti-acid rod positive and mycobacterial culture positive and biocarbium was identified as a complex group of tuberculosis.

[0029] Dramatic positive tuberculosis diagnosis only: a tuberculosis diagnosis of pulmonary tuberculosis, and at least 2 parts of sputum coating film and mycobacterium culture positive, and the strain was identified as a complex group for tuberculosis.

[0030] Molecular biological examination positive tuberculosis diagnosis: Pulmonary tuberculosis imaging performance and tuberculosis nucleic acid detection positive.

[0031] 2, Tuberculous meningitis diagnostic criteria: HIV negative, aged gr...

Embodiment 2

[0043] Example 2: DNA extraction (using the kit method DNA using the kit method, the steps are as follows)

[0044] 1. Add 0.25 g of a manure to a dry columnar tube. For fecal samples having a particularly high lipid, polysaccharide and protein content, a small amount of starting substance can increase the absorption rate and purity of DNA.

[0045] 2. Add 750 ul of PowerBead solution to the dry columnar tube.

[0046] 3. Add 60 ul of solution C1, simply flip a few or vortex.

[0047] 4, the test tube was heated at 65 ° C for 10 minutes.

[0048] 5. Use the scroll suitable for the holder to vaporize the columnar tube to the maximum speed vortex.

[0049] 6, 13000g centrifuge for 1 minute.

[0050] 7, move the centrifugal supernatant into a new 2ml collection tube. It is expected between 400-500 uL.

[0051] 8, add 250 uL solution C2 and mix even the maximum speed vortex. It was incubated for 5 minutes at 2-8 ° C.

[0052] 9,13000g centrifugation for 1 minute.

[0053] 10. Avoid co...

Embodiment 3

[0064] Example 3: Sequential sequencing analysis of intestinal flora composition (b) by 16S sequencing

[0065] 1, experimental method:

[0066] (1) After extracting genomic DNA from the sample, the conserved area of ​​RDNA is amplified with a specific primer with Barcode (Table 1 of the primer sequence information)

[0067] Table 1: Primer sequence

[0068]

[0069] (2) Then PCR amplification product cutting recovery and quantified with QuantifluorTM fluorometer. The purified amplification product is equipped with an equal amount of mixing, a sequencing joint, and the sequencing library, and the Illumina PE250 is sequenced.

[0070] 2, experimental results

[0071] (1) Analysis of the general situation of the three groups:

[0072] It was analyzed in the general case (Table 2): Health control group, tuberculous meningitis group, tuberculous meningitis group, tuberculosis group in age, gender, body mass index overall,

[0073] Table 2. Human population characteristics of HC grou...

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Abstract

The invention provides a method for screening susceptible population of tubercular meningitis or a method for predicting risk evaluation of development of pulmonary tuberculosis into tubercular meningitis. The susceptible population of tubercular meningitis in the population is judged by evaluating the abundance condition of intestinal flora of the population. Further, by detecting a fecal sample of a subject and detecting the relative abundance level of large intestine-shigella, screening of tuberculosis meningitis infection susceptibility diagnosis is carried out.

Description

Technical field [0001] The present invention relates to the field of medical diagnosis, and specific to the use of intestinal flora in the diagnosis of disease diagnosis, especially tuberculosis infectious diseases. Background technique [0002] Tuberculosis, TB) is an ancient infectious disease. It is the main cause of death of ten deaths in the world. It is also the main reason for the death of single infectious diseases. This disease usually affects the pulmonary (tuberculosis), but it can also affect other parts (extracted nuclei). In 2019, approximately 10 million people were estimated to infect tuberculosis, and about 1.2 million patients with HIV were died of tuberculosis, about 208,000 patients with HIV died of tuberculosis (Global TuberculosReport 2020.Geneva: World Health Organization ; 2020.Licence: CC BY-NC-SA3.0IGO). Pulmonary tuberculosis is also an important part of tuberculosis, and tuberculousmeningitis, TBM) is a type of mortality and disability rate in tubercul...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/06C12Q1/10
CPCC12Q1/06C12Q1/10G01N2800/26
Inventor 逄宇郭继东高孟秋刘荣梅李姗姗张福真
Owner BEIJING CHEST HOSPITAL CAPITAL MEDICAL UNIV
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