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Ready-to-use multi-probe hybridization slide, and preparation method and application thereof

A multi-probe and probe technology, applied in the field of molecular biology, can solve problems such as time-consuming and laborious, increasing the economic burden of patients, and overnight hybridization

Pending Publication Date: 2018-04-13
WUHAN HEALTHCHART BIOLOGICAL TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

If the existing hybridization technology needs to repeat multiple detections for diseases that require detection of multiple probes, it is time-consuming and laborious
In addition, traditional FISH experiments use traditional FISH probes that need to be hybridized overnight and detect multiple items at one time. Not only is it time-consuming, but the high cost also increases the financial burden on patients.

Method used

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  • Ready-to-use multi-probe hybridization slide, and preparation method and application thereof
  • Ready-to-use multi-probe hybridization slide, and preparation method and application thereof
  • Ready-to-use multi-probe hybridization slide, and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1F

[0065] The preparation of embodiment 1FISH probe

[0066] This example demonstrates the preparation process of the rapid detection probe for CBFB gene breakage, which specifically includes the following steps:

[0067] (1) Download the genome non-repeated sequence of the region covered by the CBFB gene probe from UCSC Genome Browser, where the CBFB upstream probe coverage area is: chr16:66885331-67101057, and the CBFB downstream probe coverage area is: chr16:67122220-67337946; The obtained sequence is saved in fasta format, and the repeated sequence region in the genome is replaced by N;

[0068] (2) Use the perl plug-in program chunks.pl for the genome non-repeated sequence of the region covered by the CBFB gene upstream probe obtained in step (1) (the chunks.pl program uses the code diagram see Figure 4 ) into blocks of 1kb size; import the divided blocks into OligoArray software in batches for probe design and probe screening. The conditions for probe screening are: probe...

Embodiment 2

[0076] The preparation of embodiment 2 ready-to-use multi-probe hybridization slides

[0077] The present embodiment has invented a kind of preparation method of ready-to-use multi-probe hybridization glass slide

[0078] The probe combinations used in this example were: PML / RARA, BCR / ABL, P53, D13S319, 1Q21, IGH / BCL2, CBFB, AML / ETO; in the corresponding probe attachment area. ( Figure 7 )

[0079] The hybrid slides used in this embodiment are made by entrusting the manufacturer according to the design drawing of the present invention.

[0080] The following is the preparation process of the ready-to-use multi-probe hybridization slide (M-probe Slide):

[0081] (1) According to Figure 1 ~ Figure 3 Probe-attached slides and sample-attached slides prepared by commissioned manufacturers as indicated;

[0082] (2) Carrying out probe attachment in the probe attachment region of the probe attachment glass slide;

[0083] (2.1) According to the formula: levoside 0.5% acacia ...

Embodiment 3

[0088] The method for using the ready-to-use multi-probe hybridization slide comprises the following steps:

[0089] This example demonstrates the usage method and hybridization effect of the ready-to-use multi-probe hybridization slide.

[0090] The probes included in the ready-to-use multi-probe hybridization slide used in this example are: PML / RARA, BCR / ABL, P53, D13S319, 1Q21, IGH / BCL2, CBFB, AML / ETO;

[0091] The sample detected in this embodiment is bone marrow blood lymphocytes fixed by Carnoy's fixative:

[0092] (1) Sample treatment: Add the fixed peripheral blood cells dropwise to the corresponding areas of the sample-attached slides, immerse the sample-attached slides in a container filled with 2×SSC at 37°C for 30 minutes, and pass 70% after the aging is completed. 85%, 100% gradient alcohol dehydration, dry, set aside.

[0093] (2) Probe sample covariance: Add 2ul of hybridization buffer dropwise to the probe attachment area of ​​the hybridization slide, and tur...

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Abstract

The invention belongs to the technical field of fluorescence in-situ hybridization, and particularly relates to a ready-to-use multi-probe hybridization slide, and a preparation method and applicationthereof. The multi-probe hybridization slide consists of a probe-attached slide, probes, a hybridization buffer, and a sample-attached slide. The probes are attached to mutually independent probe attachment regions respectively in a lyophilized state. A sample is dropped on a sample attachment region. When the sample attachment region is attached to a probe attachment region, the sample in the sample attachment region is hybridized with the probe of the corresponding probe attachment region. Compared with the traditional FISH experiment, multiple sets of probes can be preset on one slide at atime, and detection of multiple probes can be achieved on the same slide, in this way, the operation steps of the conventional FISH are greatly simplified, the consumption of the probes is reduced, and the using cost of the probes is reduced. The ready-to-use multi-probe hybridization slide has a very high signal-to-noise ratio, specificity, and sample detection rate.

Description

technical field [0001] The invention belongs to the field of molecular biology, and in particular relates to a ready-to-use multi-probe hybridization glass slide and a preparation method and application thereof. Background technique [0002] Fluorescence in situ hybridization is a non-radioactive molecular genetic technique developed on the basis of radioactive in situ hybridization in the 1980s. It is a new in situ hybridization method formed by replacing radioactive isotope labels with fluorescein labels. FISH has the advantages of safety, rapidity, high sensitivity and simultaneous display of multiple colors. [0003] At present, in the detection of hematology FISH, conventional slides are usually used, and the probes and slides are used separately. Usually, it is necessary to use blood samples to make slices first, and then perform a series of pretreatments on the slides. , then add the probe dropwise and seal the slide with rubber glue, and then carry out probe sample ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6841
CPCC12Q1/6841C12Q2563/107C12Q2543/10
Inventor 晏星李雪梅叶伦程弘夏陈刚
Owner WUHAN HEALTHCHART BIOLOGICAL TECH
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