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Carbohydrate chip for quickly diagnosing tuberculosis, preparation method thereof and tuberculosis diagnostic kit adopting carbohydrate chip

A technology for rapid diagnosis of tuberculosis, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of high price and limit the application and promotion of diagnostic technology, and achieve the effect of low price, simple and fast use method, and saving financial and material resources

Inactive Publication Date: 2012-10-03
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Relying on imported reagents, the price is relatively expensive, which limits the application and promotion of this diagnostic technology

Method used

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  • Carbohydrate chip for quickly diagnosing tuberculosis, preparation method thereof and tuberculosis diagnostic kit adopting carbohydrate chip
  • Carbohydrate chip for quickly diagnosing tuberculosis, preparation method thereof and tuberculosis diagnostic kit adopting carbohydrate chip
  • Carbohydrate chip for quickly diagnosing tuberculosis, preparation method thereof and tuberculosis diagnostic kit adopting carbohydrate chip

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] 1. Crude identification of mycobacterial lipopolysaccharide by silver staining

[0043] Cultivate and collect the standard strain of Mycobacterium tuberculosis H 37 Rv strain (bacterial standard strain H 37 The Rv strain was purchased from the National Institute of Pharmaceutical and Biological Products), and the bacteria were inactivated in a water bath at 100°C for 2 hours. Shake in a solvent of chloroform:methanol:water=10:10:3 for 2 hours, repeatedly degrease the cell lipids 3 times, wrap them in aluminum foil, make holes and dry them in a vacuum oven (<50°C). The dried cells were mashed with a clean glass rod, and the cells were lysed with a minimum amount of breaking buffer, and ultrasonically broken (500w, 60S×90S×12 times), and the solution after ultrasonic breaking was mixed with Triton X-114 and PBS The mixed solution was repeatedly extracted three times, the organic phases were combined, 10 times the volume of 95% ethanol was added overnight, and the precip...

Embodiment 2

[0090] Example 2 The kit was used to test the diagnostic effect of clinical tuberculosis, and it was compared with two tuberculosis antigens ESAT6 and CFP10 commonly used in the world at the same time.

[0091] 1. Specimen:

[0092] 90 cases of serum or plasma samples from clinical patients with mycobacteria (Wuhan Medical Treatment Center), divided into two groups:

[0093] Experimental group: 57 cases of active tuberculosis, all patients were diagnosed by bacteriology, imaging and dynamic observation of clinical symptoms.

[0094] Control group: non-tuberculosis patients and normal healthy people, a total of 33 cases, including pneumonia, chronic bronchopneumonia, asthma, lung cancer and normal healthy people. Non-tuberculosis patients are diagnosed according to the effects of various examination methods and clinical treatment.

[0095] 2. Method:

[0096] Use the patent kit of the present invention to detect the sera of the above-mentioned experimental group and control ...

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Abstract

The invention relates to a carbohydrate chip for quickly diagnosing tuberculosis, which comprises a solid-phase carrier. The carbohydrate chip is characterized by further comprising a mycobacterium tuberculosis lipopolysaccharides antigen probe which is fixed on the solid-phase carrier through oximation reaction. The invention also relates to a preparation method of the carbohydrate chip and a tuberculosis diagnostic kit adopting the carbohydrate chip. The preparation method comprises the following steps: separating, extracting and purifying lipopolysaccharides in a mycobacterium tuberculosis cell wall; fixing LAM (lipoarabinomannan) and LM (lipomannan) probes in the solid-phase carrier, such as a nitrocellulose membrane or porous plate, and the like, by utilizing a method of oximation reaction, thereby obtaining the carbohydrate chip; and judging if the mycobacterium tuberculosis is infected, by bonding the carbohydrate chip with a corresponding antibody in blood serum and performing enzyme linked color-developing reaction. A cell wall lipopolysaccharides antigen with species specificity adopted by the carbohydrate chip only can be combined with a specific tuberculosis antibody, so that the detecting specificity and sensitivity of the tuberculosis are greatly increased.

Description

technical field [0001] The invention relates to the field of biomedical in vitro diagnostic reagents, and more specifically, relates to a sugar chip used in the rapid diagnosis of tuberculosis, its preparation method and a tuberculosis diagnostic kit using the same. Background technique [0002] my country is a country with high tuberculosis infection and high incidence rate, and the number of tuberculosis patients ranks second in the world. The current clinical detection methods for Mycobacterium tuberculosis include bacteriological detection, serological detection and genetic diagnosis. Bacteriological detection of tuberculosis patients is a possible means of TB diagnosis, treatment and evaluation of treatment effect, which includes BACTEC detection system, mycobacterial growth indicator system, and phage reporter technology. Even after shortening the detection time, the detection method of this method still has some problems that cannot be popularized, such as the need f...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N33/543
Inventor 张舒林余晓丽孙战强
Owner SHANGHAI JIAO TONG UNIV
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