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
View PDF0 Cites 5 Cited by
  • 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 experime

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • 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...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention belongs to the technical field of fluorescence in situ hybridization, and in particular relates to a ready-to-use multi-probe hybridization glass slide and a preparation method and application thereof. The multi-probe hybridization slide is composed of a probe attachment slide, each probe, a hybridization buffer and a sample attachment slide; each probe is respectively attached to each independent probe attachment area in a freeze-dried form; The sample is dropped on the sample attachment area, and when the sample attachment area and the probe attachment area are attached, the sample in the sample attachment area is hybridized with the probe in the corresponding probe attachment area. Compared with the traditional FISH experiment, the present invention can simultaneously preset multiple groups of probes on one glass slide, and can complete the detection of various probes on the same slide, greatly simplifying the operation steps of traditional FISH; The usage amount of the probe reduces the usage cost of the probe; the ready-to-use multi-probe hybridization glass slide of the present invention has extremely 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

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
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
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap