Mycobacteria strain detecting dot blot membrane and kit

A detection kit and technology for mycobacteria, which is applied in the determination/inspection of microorganisms, microorganisms, biochemical equipment and methods, etc., and can solve problems such as easy missed detection, low pathogenicity, and small detection range

Active Publication Date: 2010-10-13
亚能生物技术(深圳)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] 3. Need special nutrients
[0019] (2) Low pathogenicity
[0020] (3) There are large differences in clinical symptoms
[0022] (5) High drug resistance rate
[

Method used

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  • Mycobacteria strain detecting dot blot membrane and kit
  • Mycobacteria strain detecting dot blot membrane and kit
  • Mycobacteria strain detecting dot blot membrane and kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0117] Embodiment 1 A kind of mycobacterium strain detection membrane strip

[0118] Comprising a substrate, and specific oligonucleotide probes immobilized on the substrate, the specific oligonucleotide probes include:

[0119] 1) A specific oligonucleotide probe for detecting Mycobacterium smegmatis, the base sequence of which is shown in SEQ ID NO: 1;

[0120] 2) A specific oligonucleotide probe for detecting Mycobacterium intracellulare, the base sequence of which is shown in SEQ ID NO: 2;

[0121] 3) a specific oligonucleotide probe for detecting Mycobacterium kansasii, the base sequence of which is shown in SEQ ID NO: 3;

[0122] 4) A specific oligonucleotide probe for detecting Mycobacterium chelonis, the base sequence of which is shown in SEQ ID NO: 4;

[0123] 5) A specific oligonucleotide probe for detecting Mycobacterium marinum, the base sequence of which is shown in SEQ ID NO: 5;

[0124] 6) A specific oligonucleotide probe for detecting Mycobacterium fortuitum...

Embodiment 2

[0143] Embodiment 2 A kind of mycobacterium strain detection kit

[0144] The kit includes: a nylon membrane immobilized with a specific oligonucleotide probe and

[0145] A primer for specifically amplifying the insertion sequence between mycobacterial 16S rRNA and 23S rRNA, the base sequence of which is shown in SEQ ID NO: 24-25;

[0146] Specific oligonucleotide probes immobilized on nylon membrane strips include:

[0147] 1) A specific oligonucleotide probe for detecting Mycobacterium smegmatis, the base sequence of which is shown in SEQ ID NO: 1;

[0148] 2) A specific oligonucleotide probe for detecting Mycobacterium intracellulare, the base sequence of which is shown in SEQ ID NO: 2;

[0149] 3) a specific oligonucleotide probe for detecting Mycobacterium kansasii, the base sequence of which is shown in SEQ ID NO: 3;

[0150] 4) A specific oligonucleotide probe for detecting Mycobacterium chelonis, the base sequence of which is shown in SEQ ID NO: 4;

[0151] 5) A s...

Embodiment 3

[0171] Clinical samples were detected with the detection kit described in Example 2,

[0172] Reagents required:

[0173] 20×SSC pH 7.0

[0174] NaCl 175.3g

[0175] Sodium citrate 88.2g

[0176] Add H 2 O 750mL was dissolved, and the pH was adjusted to 7.0 with a pH meter, and finally the volume was adjusted to 1000mL, and stored under high pressure sterilization.

[0177] 10% SDS pH 7.0

[0178] SDS 20g

[0179] Add H 2 O 180mL was dissolved, and the pH was adjusted to 7.0 with a pH meter,

[0180] Finally, the volume was adjusted to 200mL.

[0181] Liquid A (1×SSC, 0.1% SDS, pH 7.4)

[0182] 20×SSC 100mL

[0183] 10% SDS 10mL

[0184] Add H 2 O was adjusted to 1000mL.

[0185] Solution B (0.5×SSC, 0.1% SDS, pH 7.4)

[0186] 20×SSC 25mL

[0187] 10% SDS 10mL

[0188] Add H 2 O was adjusted to 1000mL.

[0189] Solution C (0.1M Sodium Citrate, pH 5.4)

[0190] 1M sodium citrate 100mL

[0191] Add H 2 O was adjusted to 1000mL.

[0192] Chromogenic solution...

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PUM

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Abstract

The invention discloses a mycobacteria strain detecting kit, comprising a dot blot membrane. A specific oligonucleotide probe for specifically detecting 23 mycobacteria and a primer for specifically amplifying an insertion sequence between mycobacteria 16S rRNA and 23S rRNA are fixed. The mycobacteria strain gene detecting kit adopts PCR combining with reverse DNA hybrid chip detection technology to simultaneously identify 23 mycobacteria within one day. The novel technology can simultaneously identify the most mycobacteria within shortest time and has the most reliable detection result. The kit distinguishes mycobacteria through the difference of gene sequences of different mycobacteria and carries out distinction and identification in the level of gene sequence of bacteria and molecular structure, so that the classification is more accurate and reliable.

Description

technical field [0001] The invention relates to the field of mycobacterium strain detection, in particular to a mycobacterium strain detection membrane strip and a kit. Background technique [0002] According to the study of the International Mycobacterial Taxonomy Working Group, mycobacteria are divided into three categories (Runyon taxonomy): [0003] 1. Slow-growing bacteria [0004] 1. Mycobacterium tuberculosis: including Mycobacterium tuberculosis human, Mycobacterium bovis, Mycobacterium africanum, Mycobacterium microti. [0005] 2. Group I photochromogenic bacteria. The characteristic is that the color of the colony is not obvious when cultured in the dark, and the colony is lemon-colored after being exposed to light for 1 hour during the proliferation period. Such as: Kansas, Mycobacterium marinum, etc. [0006] 3. Group II dark chromogenic bacteria. The characteristic is that the colonies are orange-yellow and S-shaped when cultured in the dark. Long-term exp...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12R1/34
Inventor 邹耀东任维
Owner 亚能生物技术(深圳)有限公司
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