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Detection method for identifying mycobacterium tuberculosis and non-mycobacterium tuberculosis infections

A technology of mycobacterium tuberculosis and mycobacteria, applied in the field of biomedical testing, can solve the problems of price restrictions, wide use, no simultaneous differential detection of MTB and NTM infection, low sensitivity, etc.

Inactive Publication Date: 2016-08-17
GUANGZHOU RHFAY BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, in foreign countries, based on the principle of ELISPOT, commercialized T-SPOT.TB (Oxford Immunotec, Abingdon, UK), based on the principle of ELISA QuantiFERONTB1Gold (QFT-G) and QuantiFERON-TB1Gold In-tube (QFT-IT) (Cellestis , Carnegie, Victoria, Australia) tuberculosis interferon gamma release test has been approved for use by many countries, and has been included in the LTBI and tuberculosis contact testing guidelines in many European and American countries; used, and some data show that the detection of the Chinese population has a relatively low sensitivity, and some studies suggest that this may be related to certain racial differences in the response of the population to different polypeptide fragments; at the same time, among several ELISPOT reagents studied in my country Most of the tuberculosis peptides are not self-developed
In addition, all the above-mentioned reagents are for the detection of MTB infection, and there is no ELISPOT report on simultaneous and differential detection of MTB and NTM infection

Method used

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  • Detection method for identifying mycobacterium tuberculosis and non-mycobacterium tuberculosis infections
  • Detection method for identifying mycobacterium tuberculosis and non-mycobacterium tuberculosis infections
  • Detection method for identifying mycobacterium tuberculosis and non-mycobacterium tuberculosis infections

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] [Example 1] This example includes a kit for differentiating Mycobacterium tuberculosis infection from non-tuberculous mycobacteria

[0022] Please refer to figure 1 , the kit includes detection wells, control wells and parallel control wells, wherein the detection reagents are set as:

[0023] Test 1: MTB-specific E6+E7+C14 mixed polypeptide

[0024] Detection 2: Mycobacterium-specific mixed polypeptides;

[0025] Control 1: positive control: PMA+Inomycin);

[0026] Control 2: Negative control: no peptide;

[0027] T1 and T2 are the T-SPOT.TB detection of the parallel control, respectively.

[0028] The detection steps are as follows:

[0029] 1 Isolation of mononuclear cells (PBMCs) from tuberculosis patients Ficoll lymphocyte separation medium was used to separate PBMCs, and PBMCs were resuspended in R10 culture medium (10% calf serum in RPMI1640 culture medium) for later use.

[0030] 2ELISPOT analysis Select a 96-well plate with PVDF membrane as the reaction p...

Embodiment 2

[0035] [Example 2] Test result

[0036] A small number of pulmonary tuberculosis patients (273 cases in total, including 92 cases with strain identification results and 85 cases with only sputum smear positive, totaling 187 cases of bacterium-positive patients, and 86 cases of bacterium-negative patients with negative sputum smear and sputum culture) in the laboratory The results of blood tests are shown in Table 1 and Table 2.

[0037] Table 1 Peripheral blood ELISPOT detection results of 85 cases of MTB and 7 cases of NTM positive tuberculosis

[0038]

[0039] Table 2 Peripheral blood ELISPOT detection results of 95 cases of smear-positive (bacteria-positive) pulmonary tuberculosis and 86 cases of smear-negative pulmonary tuberculosis

[0040]

[0041]The above results suggest that there is a certain proportion of NTM infection caused by both bacteria-positive and bacteria-negative pulmonary tuberculosis. Therefore, whether it is tuberculosis (bacterium-positive and...

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Abstract

The invention discloses a detection method for identifying mycobacterium tuberculosis and non-mycobacterium tuberculosis infections. The detection method comprises the following steps: respectively contacting the specific antigen of mycobacterium tuberculosis and the specific antigen of mycobacterium with a to-be-detected in-vitro T cell, and detecting a cell factor released by the T cell; comparing the detection results of the specific antigen of mycobacterium tuberculosis and the specific antigen of mycobacterium; judging that the to-be-detected in-vitro T cell is suffered from mycobacterium tuberculosis infection or non-mycobacterium tuberculosis infection according to a comparison result. According to the detection method provided by the invention, the specific antigen of mycobacterium and the ELISPOT detection method for detecting the specificity of MTB can be combined and utilized to simultaneously identify MTB and NTM infections. The detection method provided by the invention has important guidance significance on clinical diagnosis and drug usage of a doctor.

Description

technical field [0001] The invention belongs to the field of biomedical inspection, and relates to a cell immunology inspection method, in particular to a detection method for distinguishing tuberculosis mycobacterium and non-tuberculosis mycobacterium infection. Background technique [0002] Tuberculosis (TB) is one of the most common chronic infectious diseases worldwide, mainly caused by Mycobacterium tuberculosis (MTB) infection, and other mycobacteria, especially nontuberculosis mycobacteria , NTM) infection accounts for about 5% to 15% of clinical cases, and the latter has a higher proportion when AIDS is combined with tuberculosis. According to statistics, about 1 / 3 of the world's population is infected with Mtb, and 5% to 10% of those infected are tuberculosis patients. There are about 8 million to 9 million new cases of active pulmonary tuberculosis every year, and 2 million to 3 million deaths due to tuberculosis. die. [0003] Nontuberculous mycobacteria (NTM) a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569
CPCG01N33/5695G01N2333/35
Inventor 赖小敏李有生
Owner GUANGZHOU RHFAY BIOTECH CO LTD
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