Kit and detection method for detecting mycobacterium tuberculosis infection

A technology for the detection of Mycobacterium tuberculosis and infection, which is applied in the field of bioengineering, can solve the problems affecting the diagnosis and effective treatment of tuberculosis patients, the difficulty of Mycobacterium tuberculosis, and the untimely diagnosis, and achieves high specificity, easy judgment, and sensitivity. strong effect

Inactive Publication Date: 2017-08-18
THE FIRST AFFILIATED HOSPITAL OF XINXIANG MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, most of the current detection samples for Mycobacterium tuberculosis are aimed at sputum samples, and most of the sputum samples are collected by patients themselves. Human errors are likely to cause specimen infection and affect the accuracy of detection.
And because of the early stage of tuberculosis infection, it is difficult to accurately detect Mycobacterium tuberculosis in the blood, which may easily lead to untimely diagnosis and affect the diagnosis and effective treatment of tuberculosis patients.

Method used

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  • Kit and detection method for detecting mycobacterium tuberculosis infection
  • Kit and detection method for detecting mycobacterium tuberculosis infection
  • Kit and detection method for detecting mycobacterium tuberculosis infection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] The invention provides a test kit for detecting Mycobacterium tuberculosis infection, comprising the following parts:

[0038] It consists of a detection primer, a 10-fold loop-mediated isothermal amplification reaction solution, a DNA polymerase, a positive control, a negative control, a chromogenic agent, and a cell expansion solution; the detection primer is composed of a pair of outer primers F3 / B3 and A pair of internal primers FIP / BIP consists of:

[0039] The sequence of the outer primer F3 is: 5'-CTGGGCCTCAAGCAAGG-3' (SEQ ID NO.1)

[0040] The sequence of the outer primer B3 is: 5'-CGACGTCGCTGACCACTT-3' (SEQ ID NO.2)

[0041] The sequence of the inner primer FIP is (SEQ ID NO.3):

[0042] 5'-CGGGCGACGGTTTCGAAGTCGGAAGACGGGGTCAACGGT-3'

[0043] The sequence of the inner primer BIP is (SEQ ID NO.4):

[0044]5'-CGGCTGCAAGAGATGGCGTCTTGGGTCACCCTCTCGT-3'

[0045] Wherein, the molar ratio of the pair of outer primers F3 / B3 to the pair of inner primers FIP / BIP is 1:...

Embodiment 2

[0051] The invention provides a test kit for detecting Mycobacterium tuberculosis infection, comprising the following parts:

[0052] It consists of a detection primer, a 10-fold loop-mediated isothermal amplification reaction solution, a DNA polymerase, a positive control, a negative control, a chromogenic agent, and a cell expansion solution; the detection primer is composed of a pair of outer primers F3 / B3 and Composed of a pair of internal primers FIP / BIP,

[0053] The sequence of the outer primer F3 is: 5'-CTGGGCCTCAAGCAAGG-3' (SEQ ID NO.1)

[0054] The sequence of the outer primer B3 is: 5'-CGACGTCGCTGACCACTT-3' (SEQ ID NO.2)

[0055] The sequence of the inner primer FIP is (SEQ ID NO.3):

[0056] 5'-CGGGCGACGGTTTCGAAGTCGGAAGACGGGGTCAACGGT-3'

[0057] The sequence of the inner primer BIP is (SEQ ID NO.4):

[0058] 5'-CGGCTGCAAGAGATGGCGTCTTGGGTCACCCTCTCGT-3'

[0059] Wherein, the molar ratio of the pair of outer primers F3 / B3 and a pair of inner primers FIP / BIP is 1:...

Embodiment 3

[0065] The invention provides a test kit for detecting Mycobacterium tuberculosis infection, comprising the following parts:

[0066] It consists of a detection primer, a 10-fold loop-mediated isothermal amplification reaction solution, a DNA polymerase, a positive control, a negative control, a chromogenic agent, and a cell expansion solution; the detection primer is composed of a pair of outer primers F3 / B3 and Composed of a pair of internal primers FIP / BIP,

[0067] The sequence of the outer primer F3 is: 5'-CTGGGCCTCAAGCAAGG-3' (SEQ ID NO.1)

[0068] The sequence of the outer primer B3 is: 5'-CGACGTCGCTGACCACTT-3' (SEQ ID NO.2)

[0069] The sequence of the inner primer FIP is (SEQ ID NO.3):

[0070] 5'-CGGGCGACGGTTTCGAAGTCGGAAGACGGGGTCAACGGT-3'

[0071] The sequence of the inner primer BIP is (SEQ ID NO.4):

[0072] 5'-CGGCTGCAAGAGATGGCGTCTTGGGTCACCCTCTCGT-3'

[0073] Wherein, the molar ratio of the pair of outer primers F3 / B3 and a pair of inner primers FIP / BIP is 1:...

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Abstract

The invention discloses a kit for detecting mycobacterium tuberculosis infection. The kit comprises the following components: a detection primer, a 10-fold loop-mediated isothermal amplification reaction liquid, a DNA polymerase, a positive control, a negative control, a color developing agent and a bacteria spreading cultivation liquid, wherein the detection primer comprises a pair of outer primers F3 / B3 and a pair of inner primers FIP / BIP. The kit is capable of efficiently, quickly, accurately and sensitively obtaining the detection result of mycobacterium tuberculosis infection in blood, the detection result is high in specificity and high in sensitivity, result judgment can be easily completed with eyes instead of electrophoresis, and the kit is very suitable for fast detection of mycobacterium tuberculosis.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and in particular relates to a kit and a detection method for detecting mycobacterium tuberculosis infection. Background technique [0002] Tuberculosis caused by Mycobacterium tuberculosis (Mtb) is an infectious disease that seriously endangers human health. In recent decades, due to population movement, the spread of drug-resistant tuberculosis has become an important cause of all infectious diseases. If it is not effectively controlled, more people may develop tuberculosis in the future. [0003] The interferon-gamma (IFN-γ) release assay (IGRA) in the new immunological method is a method with high accuracy for detecting Mycobacterium tuberculosis infection at this stage. The existing commercial kits include T-SPOT .TB and QuantiFERON-TBGoldIn-Tube (QFT-IT), the sensitivity and specificity of these two kits are between 80-90% respectively, which needs to be improved. [0004] Loop-me...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/04C12R1/32
CPCC12Q1/6844C12Q1/689C12Q2531/119
Inventor 张边防崔俊伟高渐联周爱杰李振云
Owner THE FIRST AFFILIATED HOSPITAL OF XINXIANG MEDICAL UNIV
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