CN107904283A - A kind of instant multiprobe hybridized slides and its preparation method and application - Google Patents
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Abstract
The invention belongs to fluorescence in situ hybridization technique field, and in particular to a kind of instant multiprobe hybridized slides and its preparation method and application.The multiprobe hybridized slides are made of probe attachment slide, each probe, hybridization buffer and sample attachment slide；Each probe is respectively attached on each mutually independent probe attachment region in the form of lyophilized；Sample drop is added on sample attachment region, and when sample attachment region is bonded with probe attachment region, the probe of the sample of sample attachment region and corresponding probe attachment region realizes that hybridization combines.Tested compared to traditional FISH, the present invention can further open preset multigroup probe, and can complete the detection of a variety of probes on same slide again at the same time on slide, enormously simplify the operating procedure of traditional FISH；Reduce the use cost that the usage amount of probe reduces probe；Instant multiprobe hybridized slides of the present invention have high signal-to-noise ratio, specificity and sample recall rate.
Description
Technical field
The invention belongs to biology field, and in particular to a kind of instant multiprobe hybridized slides and preparation method thereof
And application.
Background technology
Fluorescence in situ hybridization be it is a kind of grow up in the 1980s radioactive in situ hybridization technical foundation one
Kind of on-radiation molecular genetic techniques, what is marked substitution labelled with radioisotope with fluorescein and formed is a kind of new in situ miscellaneous
Friendship method.FISH has the advantages that safety, quick, high sensitivity and shows multiple color at the same time.
At present in the detection of hematology FISH, usually used is all conventional slide, and probe and glass slide are
Separately it is used alone, it usually needs first using blood sample film-making, then slide is subjected to a series of pre-treatments, then probe is added dropwise
And rubber glue mounting is used, the co-variation of probe sample and hybridization are carried out again after mounting；Blood disease such as leukaemia common at present
FISH detection projects up to tens kinds of number, such as CLL (chronic myelocytic leukemia) needs to detect cMYC gene breaks, P16
Missing, E2A fractures, hyperdiploid detection (CEP10/CEP17/CEP4), TEL/AML1 fusions, MLL fractures, BCR/ABL fusions
And disease type and process are assessed in IGH fractures etc..If with existing hybridization technique for needing to detect a variety of probes
Need to repeat repeated detection during disease, it is time-consuming and laborious.In addition traditional FISH experiments use traditional FISH probe to need
Want overnight hybridization and disposably detect multiple projects not only to take, but also high expense also increases the economy of patient and bears
Load.
The content of the invention
The present invention is in view of the deficiencies of the prior art, and it is an object of the present invention to provide a kind of instant multiprobe hybridized slides and its preparation
Methods and applications.
For achieving the above object, the technical solution adopted by the present invention is：
A kind of instant multiprobe hybridized slides kit, by probe attachment slide, each probe, hybridization buffer and sample
Adhere to slide composition；Include several mutually independent probe attachment regions on probe attachment slide, each probe is to freeze
Form is respectively attached on each mutually independent probe attachment region；It is mutually independent comprising several on the sample attachment slide
Sample attachment region, sample attachment region are corresponded with probe attachment region, and sample drop is added on sample attachment region, when sample attachment region
When being bonded with probe attachment region, the probe of the sample of sample attachment region and corresponding probe attachment region realizes that hybridization combines.
In such scheme, the probe attachment slide is connect by detecting probe information tag slot, probe adhering zone and hybridized slides
Close district's groups into, corresponding probe species on each independent probe attachment region are have recorded on the detecting probe information tag slot, it is described
Several mutually independent probe attachment regions are included in probe adhering zone, probe is attached to probe attachment region in the form of lyophilized
On, the surface of the hybridized slides bonding land has acrylate glue, for probe attachment slide packaging sealer and and
Sample adheres to the bonding of slide.
In such scheme, the probe adhering zone is a groove, and groove includes several mutually independent probe attachments
Block of cells, each independent probe attachment block of cells are made of probe attachment region and peripheral isolated area；Each independent probe is attached
Between block of cells and there is isolation ditch.
In such scheme, the sample attachment slide isolates district's groups by sample message recording areas, sample attachment region and sample
Into have recorded sample information on the sample message recording areas, sample drop is added on sample attachment region, and the sample isolated area is used
In preventing sample cross contamination.
In such scheme, the sample attachment region uses hydrophilic material；The sample isolated area has hydrophobic coating.
In such scheme, the component of the hybridization buffer is：10wt% ethylene carbonates, 30wt% dextran sulfates,
1ugRNaseA, 0.6M sodium chloride, 10mM citrate buffer solutions.
In such scheme, the probe is FISH gene locis probe or FISH genes centromere/telomere probe.
In such scheme, each probe is selected from following combination A or combination B：
Combine A：BCR/ABL, IGH/BCL2, AML/ETO；
Combine B：PML/RARA, BCR/ABL, P53, D13S319,1Q21, IGH/BCL2, CBFB, AML/ETO.
The preparation method of above-mentioned instant multiprobe hybridized slides, includes the following steps：
(1) the probe attachment slide for being attached with each probe is prepared：
A. the probe attachment slide with said structure is prepared；
B. excipient is prepared using deionized water, the formula of the excipient is：Left-handed glucosides 0.5wt%, Arabic gum
Set 3wt%, mannitol 1wt%, polyacrylate 0.3wt%；
C. each probe solution is prepared respectively, each probe solution is uniformly mixed with excipient respectively, is added dropwise respectively and uniform
It is applied on probe attachment slide on each independent probe attachment region；Then probe attachment slide is freezed, sealer, be prepared into
Adhere to slide to the probe for being attached with each probe；
(2) the sample attachment slide with said structure is prepared；
(3) preparing hybrid buffer solution；
(4) the probe attachment slide, hybridization buffer and sample attachment slide that probe is attached with by preparing gained form i.e.
With type multiprobe hybridized slides.
The application of above-mentioned instant multiprobe hybridized slides, specifically comprises the following steps：
(1) sample process：The peripheral blood cells fixed are added drop-wise to the sample attachment region of sample attachment slide respectively, will
Sample attachment slide immerses aging 30min in the container for filling 37 DEG C of 2XSSC, and the high fire of micro-wave oven heats 3min until liquid
Boiling, then again in low fire continue heat 10min；Slide is placed in -20 refrigerator degree Celsius precoolings immediately after the completion of processing
Graded ethanol in be dehydrated and dry, it is spare；
(2) probe sample co-variation：Hybridization buffer is added dropwise to probe attachment region on probe attachment slide respectively, and by sample
This attachment slide is buckled to, and sample attachment region is fit together with probe attachment region, by whole hybridized slides and sample after fitting
Attachment slide inverts together, is placed in 73 DEG C of denaturation 5min of metal bath, hybrid device is placed in 37 DEG C of wet box hybridization after the completion of deformation
1h；
(3) post-hybridization washing：Wait to remove probe attachment slide after the completion of hybridizing, sample attachment slide is placed in 60 DEG C of preheatings
Washing lotion (sodium chloride containing 0.3M, 0.03M sodium citrates and 0.1%Tween-20) in washing remove uncombined probe；Wash
Slide is placed in room temperature distilled water after and is washed again, and room temperature is dried；
(4) redye and microscopy：Anti-cancellation mountant is added dropwise to the sample attachment region of the sample attachment slide dried, uses lid
Fragmentation mounting and in fluorescence microscopy Microscopic observation results of hybridization.
The preparation method of heretofore described FISH gene locis probe is as follows：
(1) the genome non repetitive sequence of probe institute overlay area is downloaded from UCSC Genome Browser, by acquisition
The genome non repetitive sequence of probe institute overlay area is divided into the block of 1kb sizes using perl plug-in card programs chunks.pl；
Block batch after segmentation is imported into OligoArray softwares and carries out probe design, probe screening, the probe that then will be filtered out
Export in Microsoft Excel, then add the sequence label of 18bp with 3 ' ends at 5 ' ends of every probe respectively, obtain a series of
Probe sequence with sequence label；
(2) chemical synthesis is carried out to the probe sequence with sequence label obtained by step (1) using DNA synthesizer, will changed
The probe for learning synthesis is mixed, and probe library is prepared；It is respectively synthesized 5 ' M13 universal primers of the end with green fluorescence group
With 5 ' M13 universal primers of the end with fluorophor, probe library is carried out using 5 ' M13 universal primers of the end with fluorophor
Amplification mark reaction；
(3) the amplification marked product of probe library obtained by step (2) is purified, dilute after obtain the probe library of fluorescent marker.
In such scheme, the condition of the screening of probe described in step (1) is：85~99 DEG C of probe length 50bp, TM value,
GC is than 40~80%, without TTTT/GGGG/AAAA/CCCC, minimum interval 5bp between probe；Step (1) described sequence label
Base sequence is as follows：
5 ' end sequence labels be：TGTAAAACGACGGCCAG (M13 upstream sequences)
3 ' end sequence labels be：GGTCATAGCTGTTTCCTG (M13 downstream sequences).
In such scheme, universal primer sequence is described in step (2)：M13-F TGTAAAACGACGGCCAGT； M13-
R CAGGAAACAGCTATGACC。
In such scheme, the PCR reaction systems of step (2) the amplification mark reaction are as follows：
In such scheme, the PCR reaction conditions of step (2) the amplification mark reaction are as follows：95℃5min；94 DEG C of 30s,
58 DEG C of 30s, 72 DEG C of 30s, carry out 20 circulations altogether；72℃10min.
The preparation method of above-mentioned FISH gene locis probe is suitable for the preparation of FISH probe set forth below, specifically includes：
ALK、 BCL2、BCL3、BCL6、BCL10、BCL12、BCR、CCND1、E2A、EGFR、ETV6、FIP1L1、 HER2、IGH、IGK、
IGL、MALT1、MLL(ALL-1、HTRX1、HRX)、MYC(c-Myc)、PAX5、 PDGFRA、PDGFRB、SIL、TCF3(E2A、
ITF1), TCL1A, TCRAD, TCRB, TCRG, TLX1, TLX3 (HOX11L2, RNX) or TOP2A, BASE, BRCA1, CCND1,
CCNE1、DCD、E2F3、 n-MYC/MYCN、COX-2/PTGS2、LRIG1、ERa、hTERT、MLN64/STARD3、PGR、
SNAI1、 SRC、TOP1、TUBB1、AIB1、DLC-1、EDD、Pip4k2b/5k、Sil、TBX2、c-Kit、VEGF、VCAM-1、
Tie-1、Ts/TYMS、PSMA、PSA、PAP、P15、P16、BCL1、BCL2、MTOR、TIMP1、ESR1、 PTEN、MDM2/CDK4、
MET、C-MET、ERB1、FGFR1、IGF1R、NET、FGFR3、ABCB1、 TMPRSS2、BRCA2、TOP2B、ERCC1、AKT1、
AKT2、AKT3、HRAS、NRAS、RAF1、HER3、 HER4、ENT1、RRM1、RRM2、RRM2B、PIK3CA、AURK4、AURKB、
AURKC、MAPT/tau、 TTBK1、TUBB、VEGFR、CCND3、CDK6、CDK2、CDC2、HDAC、ESR2、SCUBE2、BIRC5、
FASN、DHFR、TP/ECGF1、TYMP、DPYD、TK1、HMGIC、ABCA2、ABCB11、ABCC1、 ABCC2、ABCC3、ABCC4、
ABCC5、ABCG2、MVP、ATP7A、ATP7B、SLC29A1、SLC28A1、 SLC19A1、TUBB4、TUBA、MAP4、MAP7、
STMN1、KIF5B、HSPA5、PSMD14、FPGS、 GSTP1、GPX、GCLC、：ABL t(9；22)(q34；q11)、PRDM16del
(1p36.32)del(21q22.12)、 RUNX1/AML1del(1p36.32)del(21q22.12)、CEP8、PDGFRB、
NUP98、FGFR1、ASS、ETO t(8； 21)(q22；q22)、AML1t(8；21)(q22；q22)、CBFbeta inv(16)
(p13q22)t(16；16)(p13；q22)、 MYH11inv(16)(p13q22)t(16；16)(p13；q22)、AF9t(9；11)、
PML t(15；17)(q22；q21)、 PLZF t(11；17)(q23；q21)、NuMAt(11；17)(q13；q21)、NPMt(5；
17)(q23；q12)、RAR αt(15；17)(q22；q21)t(11；17)(q23；q21)t(11；17)(q13；q21)t(5；17)
(q23；q21)、 EVI1t(3；v)(q26；v).
The preparation method of heretofore described FISH genes centromere and telomere probe is as follows：
(1) specific fragment of PCR amplification centromeric probe or telomere probe；
(2) nick-translation method marks above-mentioned specific fragment (mark system is conventional nick translation body)；
(3) using the prepared centromere/telomere probe of acetate ethanol sodium method purifying, using deionized water dissolving and will visit
Pin concentration dilution is spare for 100ng/ul.
The preparation method of the FISH genes centromere and telomere probe is suitable for centromere/telomere probe listed below
Prepare, specifically：CEP1、CEP2、CEP3、CEP4、CEP5、CEP6、CEP7、CEP8、CEP9、CEP10、CEP11、 CEP12、
CEP13、CEP14、CEP15、CEP16、CEP17、CEP18、CEP19、CEP20、CEP21、CEP22、 CEP23、CEP X、CEP
Y and telomere.
In the present invention, probe used using pipettor is applied to corresponding independent spy after being dissolved using vehicle composition
Pin attachment region.Specifically excipient composition composition formula is：Left-handed glucosides 0.5%, acacia 3%, mannitol 1%, poly- third
Olefin(e) acid ester 0.3%, using deionized water dissolving, 4 DEG C save backup.
Multiprobe hybridized slides of the present invention need to adhere to each probe attachment region of slide when specifically used in probe
2 μ l hybridization buffers are added dropwise, then tip upside down on sample attachment slide on probe attachment slide, sample and probe combine, and will tie
Hybridized slides after conjunction are placed in 73 DEG C of denaturation 2min on metal bath, are placed in 37 DEG C of wet box hybridization 2h.
Beneficial effects of the present invention：
(1) tested compared to traditional FISH, the present invention can further open preset multigroup probe, and can at the same time on slide
To complete the detection of a variety of probes on same slide again, the operating procedure of traditional FISH enormously simplify；
(2) compared with existing all technologies the present invention by unique design, and coordinate specific hybridization buffer with
And unique detection method, reduce the use cost that the usage amount of probe reduces probe；
(3) instant multiprobe hybridized slides of the present invention have high signal-to-noise ratio, specificity and sample inspection
Extracting rate；
(4) compared to the preparation method for the rapid fluorescence in situ hybridization probe without repetitive sequence established in recent years, sheet
The probe screening that invention uses relies primarily on program completion, and uses more stable PCR methods amplification and label probe,
And probe mark rate higher is more homogeneous, fluorescent marker is only carried out at 5 end of probe so not influencing the hybridization match reaction of probe；
In addition the present invention does not use the nick translation easily influenced by environment or operation technique to react and random primer reacts to mark spy
Pin, the probe length marked is consistent, and mark yield is high and adds the stability between batch.
Brief description of the drawings
Fig. 1 is that the probe of multiprobe hybridized slides of the present invention adheres to slide plan.
Fig. 2 is that the probe of multiprobe hybridized slides of the present invention adheres to slide stereochemical structure and dimension data.
Fig. 3 is that the sample of multiprobe hybridized slides of the present invention adheres to slide plan.
Wherein, 1 is detecting probe information tag slot, and 2 be probe adhering zone, and 3 be hybridized slides bonding land, and 4 adhere to for probe
Area, 5 be peripheral isolated area, and 6 be isolation ditch, and 7 be sample message recording areas, and 8 be sample attachment region, and 9 be sample isolated area.
Fig. 4 is that chunks.pl programs use code map in embodiment 1.
Fig. 5 is that CBFB gene break detection kits part probe sequence figure after repetitive sequence is removed in embodiment 1.
Fig. 6 prepares schematic diagram for probe.
Fig. 7 is instant multiprobe hybridized slides schematic diagram described in embodiment 3.
Fig. 8 is testing result figure of the present invention.
Embodiment
For a better understanding of the present invention, with reference to the embodiment content that the present invention is furture elucidated, but the present invention
Content is not limited solely to the following examples.
Such as attached drawing 1~3, a kind of multiprobe hybridized slides, by probe attachment slide (A) and sample attachment slide (B) two
Divide and form.The probe attachment slide (A) is by detecting probe information tag slot (1), probe adhering zone (2) and hybridized slides bonding land
(3) form, the probe adhering zone (2) is a groove, and groove includes several mutually independent probe attachment block of cells, often
A independent probe attachment block of cells is made of probe attachment region (4) and peripheral isolated area (5), and each independent probe attachment is small
There is isolation ditch (6) between block, probe is attached on probe attachment region in the form of lyophilized；The detecting probe information tag slot
(1) corresponding probe species on each independent probe attachment region, the surface of the hybridized slides bonding land (3) be have recorded on
With acrylate glue, the bonding for the packaging sealer of probe attachment slide and with sample attachment slide.
The sample attachment slide (B) is by sample message recording areas (7), several mutually independent sample attachment regions (8)
Formed with sample isolated area (9), sample drop is added on sample attachment region (8), and the sample attachment region uses hydrophilic material, described
Sample isolated area has hydrophobic coating, and sample information, sample attachment region (8) and probe are have recorded on the sample message recording areas
Attachment region (4) is buckled to when on probe attachment slide, sample and the phase of sample attachment region in corresponding when sample adheres to slide
The probe of corresponding probe attachment region combines, and then realizes hybridization.
The preparation of embodiment 1FISH probes
Embodiment shows the preparation process of the quick detection probe of CBFB gene breaks, specifically comprise the following steps：
(1) the genome non repetitive sequence of CBFB gene probes institute overlay area is downloaded from UCSC Genome Browser,
Wherein CBFB upstream probes overlay area is：chr16:66885331-67101057, CBFB downstream probe overlay area are：
chr16:67122220-67337946；Acquired sequence saves as fasta forms, and repetitive sequence region is replaced in genome
For N；
(2) the genome non repetitive sequence for the CBFB upstream region of gene probe institute overlay area for obtaining step (1) uses
Perl plug-in card programs chunks.pl (chunks.pl programs are shown in Fig. 4 using code map) is divided into the block of 1kb sizes；It will divide
Block batch after cutting imports OligoArray softwares and carries out probe design, probe screening, and the condition of probe screening is：Probe is grown
50bp is spent, 85~99 DEG C of TM values, GC is than 40~80%, without TTTT/GGGG/AAAA/CCCC, minimum interval 5bp between probe；So
The probe filtered out is exported in Microsoft Excel afterwards, then the label at 5 ' ends of every probe with 3 ' ends plus 18bp respectively
Sequence, obtaining a series of CBFB upstream region of gene probe sequences with label, (Fig. 5 show CBFB gene probes pond, due to sequence
Row are excessive not to be shown one by one)；The base sequence of sequence label is as follows：5 ' end sequence labels be： TGTAAAACGACGGCCAG
(M13 upstream sequences), 3 ' end sequence labels are：GGTCATAGCTGTTTCCTG (M13 downstream sequences)；
(3) the genome non repetitive sequence for the CBFB downstream of gene probe institute overlay area for obtaining step (1) uses
Perl plug-in card programs chunks.pl (chunks.pl programs are shown in Fig. 4 using code map) is divided into the block of 1kb sizes；It will divide
Block batch after cutting imports OligoArray softwares and carries out probe design, probe screening, and the condition of probe screening is：Probe is grown
50bp is spent, 85~99 DEG C of TM values, GC is than 40~80%, without TTTTT/GGGG/AAAAA/CCCC, minimum interval between probe
5bp；Then the probe filtered out is exported in Microsoft Excel, then adds 18bp with 3 ' ends at 5 ' ends of every probe respectively
Sequence label (base sequence is same as above), obtain a series of CBFB downstream of gene probe sequences with label；
(4) using DNA synthesizer to the CBFB upstream region of gene probe sequences with sequence label obtained by step (2)
Synthesis is learned, the probe of chemical synthesis is mixed, CBFB upstream region of gene probe libraries are prepared, the CBFB upstream region of gene is visited
Pin storehouse includes 1109 CBFB upstream region of gene probes for carrying sequence label；Similarly, using DNA synthesizer to obtained by step (3)
CBFB downstream of gene probe sequence with sequence label carries out chemical synthesis, and the probe of chemical synthesis is mixed, and prepares
CBFB downstream of gene probe libraries are obtained, the CBFB downstream of gene probe library includes 1238 CBFB genes for carrying sequence label
Downstream probe；
(5) M13 universal primer of the 5 ' ends with green fluorescence group and Chinese red fluorophor, universal primer are respectively synthesized
Sequence is：M13-F TGTAAAACGACGGCCAGT, M13-R CAGGAAACAGCTATGACC；Green fluorescence is carried using 5 ' ends
The M13 universal primers of group carry out CBFB upstream region of gene probe library amplification mark reaction；Use 5 ' fluorescence of the end with Exocarpium Citri Rubrum
The M13 universal primers of group carry out CBFB downstream of gene probe library amplification mark reaction；The PCR of the amplification mark reaction is anti-
Answer system as follows：
The PCR reaction conditions of the amplification mark reaction are as follows：95℃5min；94 DEG C of 30s, 58 DEG C of 30s, 72 DEG C of 30s, altogether
Carry out 20 circulations；72℃10min.
(6) using acetate ethanol sodium method respectively to the amplification marked product of CBFB upstream region of gene probe libraries obtained by step (5)
Purified with the amplification marked product of CBFB downstream of gene probe libraries, obtained again through dilution after purification：Concentration is 100ng/ul
Fluorescent marker CBFB upstream region of gene probe library and concentration be 100ng/ul fluorescent marker CBFB downstream of gene probe libraries.
The preparation of 2 instant multiprobe hybridized slides of embodiment
The present embodiment has invented a kind of preparation method of instant multiprobe hybridized slides
Probe combinations used in the present embodiment are respectively：PML/RARA、BCR/ABL、P53、D13S319、1Q21、
IGH/BCL2、CBFB、AML/ETO；Every group of probe matches somebody with somebody manufacturing probe excipient respectively, is applied to corresponding probe attachment region.(figure
7)
Hybridized slides used are manufactured according to design drawing of the present invention commission manufacturer in the present embodiment.
It is the preparation process of instant multiprobe hybridized slides (M-probe Slide) below：
(1) according to being entrusted shown in Fig. 1~Fig. 3, manufacturer prepares probe attachment slide and sample adheres to slide；
(2) probe attachment is carried out in the probe attachment region of probe attachment slide；
(2.1) according to formula：3% mannitol of left-handed 0.5% acacia of glucosides, 1% polyacrylate 0.3%, uses
The above-mentioned vehicle composition of deionized water dissolving, 4 DEG C save backup；
(2.2) each probe compositions are prepared：Probe aqueous solution final concentration is 100ng/ul；
(2.3) 10ul probe compositions and 90ul vehicle compositions are drawn, is uniformly mixed, 5ul is drawn using pipettor
Mixed probe vehicle composition is added dropwise and is uniformly applied to each probe attachment region；
(2.4) probe for being attached with probe vehicle composition attachment slide is put into freeze dryer and freezed, use is special
It is spare after sealer sealer.Pay attention to:Lucifuge is answered in operating process
Embodiment 3
The application method of instant multiprobe hybridized slides, comprises the steps of：
Embodiment shows the application method and crossbreeding effect of instant multiprobe hybridized slides.
The probe that instant multiprobe hybridized slides used in the present embodiment include is：PML/RARA、BCR/ABL、P53、
D13S319、1Q21、IGH/BCL2、CBFB、AML/ETO；
The sample that the present embodiment is detected is the marrow blood lymphocyte after the fixation of Ka Nuoshi fixers：
(1) sample process：The peripheral blood cells fixed are added drop-wise to the corresponding region of sample attachment slide respectively, by sample
This attachment slide immerses aging 30min in the container for filling 37 DEG C of 2 × SSC, 70%, 85% is crossed after the completion of aging, 100% gradient
Dehydration of alcohol dries, spare.
(2) probe sample co-variation：2ul hybridization buffers are added dropwise to the probe attachment region of hybridized slides respectively, and by sample
This attachment slide is buckled to, and sample attachment region is fit together with probe attachment region, by whole hybridized slides and sample after fitting
Attachment slide inverts together, is placed in 73 DEG C of denaturation 5min of metal bath, hybrid device is placed in 37 DEG C of wet box hybridization after the completion of deformation
1h。
(3) post-hybridization washing：Wait to remove hybridized slides after the completion of hybridizing, the washing lotion that slide is placed in 60 DEG C of preheatings (contains
0.3M sodium chloride, 0.03M sodium citrates and 0.1%Tween-20) in washing remove uncombined probe.By glass after the completion of washing
Piece is placed in room temperature distilled water and washs again, and room temperature is dried.
(4) redye and microscopy：5ul anti-cancellation mountants are added dropwise per region to the sample attachment slide corresponding region dried,
Using suitable coverslip mounting and in fluorescence microscopy Microscopic observation results of hybridization；The results are shown in Figure 8.
As can be seen from Figure 8：The present invention is used for having high noise with outstanding crossbreeding effect when multiprobe hybridizes
Than, specificity and sample recall rate.
Obviously, above-described embodiment is only intended to clearly illustrate made example, and is not the limitation to embodiment.It is right
For those of ordinary skill in the art, can also make on the basis of the above description it is other it is various forms of change or
Change.There is no necessity and possibility to exhaust all the enbodiments.And the obvious change or change therefore amplified
Move within still in the protection domain of the invention.
Claims (10)
1. a kind of instant multiprobe hybridized slides kit, it is characterised in that slow by probe attachment slide, each probe, hybridization
Fliud flushing and sample attachment slide composition；Several mutually independent probe attachment regions are included on the probe attachment slide, it is each to visit
Pin is respectively attached on each mutually independent probe attachment region in the form of lyophilized；Several are included on the sample attachment slide
Mutually independent sample attachment region, sample attachment region are corresponded with probe attachment region, and sample drop is added on sample attachment region, when
When sample attachment region is bonded with probe attachment region, the probe of the sample of sample attachment region and corresponding probe attachment region is realized miscellaneous
Knot is closed.
2. instant multiprobe hybridized slides kit according to claim 1, it is characterised in that the probe adheres to slide
It is made of detecting probe information tag slot, probe adhering zone and hybridized slides bonding land, have recorded on the detecting probe information tag slot
Corresponding probe species on each independent probe attachment region, the probe adhering zone is interior to include several mutually independent spies
Pin attachment region, probe are attached on probe attachment region in the form of lyophilized, and the surface of the hybridized slides bonding land has propylene
Sour fat glue, the bonding for the packaging sealer of probe attachment slide and with sample attachment slide.
3. instant multiprobe hybridized slides kit according to claim 2, it is characterised in that the probe adhering zone
For a groove, groove includes several mutually independent probe attachment block of cells, and each independent probe adheres to block of cells by visiting
Pin attachment region and peripheral isolated area form；There is isolation ditch between each independent probe attachment block of cells.
4. instant multiprobe hybridized slides kit according to claim 1, it is characterised in that the sample adheres to slide
It is made of sample message recording areas, sample attachment region and sample isolated area, sample letter is have recorded on the sample message recording areas
Breath, sample drop are added on sample attachment region, and the sample isolated area is used to prevent sample cross contamination.
5. instant multiprobe hybridized slides kit according to claim 4, it is characterised in that the sample attachment region is adopted
With hydrophilic material；The sample isolated area has hydrophobic coating.
6. instant multiprobe hybridized slides kit according to claim 1, it is characterised in that the hybridization buffer
Component is：10wt% ethylene carbonates, 30wt% dextran sulfates, 1ugRNaseA, 0.6M sodium chloride, 10mM citrate buffer solutions.
7. instant multiprobe hybridized slides kit according to claim 1, it is characterised in that each probe is FISH
Gene loci probe or FISH genes centromere/telomere probe.
8. instant multiprobe hybridized slides kit according to claim 7, it is characterised in that each probe is selected from such as the following group
Close A or combination B：
Combine A：BCR/ABL, IGH/BCL2, AML/ETO；
Combine B：PML/RARA, BCR/ABL, P53, D13S319,1Q21, IGH/BCL2, CBFB, AML/ETO.
9. the preparation method of any instant multiprobe hybridized slides of claim 1 ~ 8, it is characterised in that including following step
Suddenly：
（1）Prepare the probe attachment slide for being attached with each probe：
A. the probe attachment slide with any structure of claim 1 ~ 8 is prepared；
B. excipient is prepared using deionized water, the formula of the excipient is：Left-handed glucosides 0.5wt%, acacia
3wt%, mannitol 1wt%, polyacrylate 0.3wt%；
C. each probe solution is prepared respectively, each probe solution is uniformly mixed with excipient respectively, is added dropwise and is uniformly smeared respectively
On probe attachment slide on each independent probe attachment region；Then probe attachment slide is freezed, sealer, be prepared attached
The probe attachment slide of each probe；
（2）Prepare the sample attachment slide with any structure of claim 1 ~ 8；
（3）Preparing hybrid buffer solution；Probe attachment slide, hybridization buffer and the sample that each probe is attached with obtained by preparing are attached
Slide composition instant multiprobe hybridized slides.
10. the application of any instant multiprobe hybridized slides of claim 1 ~ 8, specifically comprises the following steps：
（1）Sample process：Sample is added drop-wise to each sample attachment region of sample attachment slide respectively, it is after pretreatment, spare；
（2）Probe sample co-variation：Hybridization Buffer is added dropwise to each mutually independent probe attachment region on probe attachment slide respectively
Liquid, and sample attachment slide is tipped upside down on probe attachment slide, sample attachment region is fit together with probe attachment region, paste
Whole probe attachment slide is inverted together with sample attachment slide after conjunction, is placed in 73 DEG C of denaturation 5min of metal bath, deformation is completed
It is placed in 37 DEG C of wet box hybridization 1h again afterwards；
（3）Post-hybridization washing：Wait to remove probe attachment slide after the completion of hybridizing, room temperature after sample attachment slide washing is dried；
（4）Redye and microscopy：Anti-cancellation mountant is added dropwise to the sample attachment region of the sample attachment slide dried, uses lid fragmentation
Mounting and in fluorescence microscopy Microscopic observation results of hybridization.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108709991A (en) * | 2018-06-20 | 2018-10-26 | 浙江天杭生物科技股份有限公司 | A kind of slide and its application method of bovine viral Immunofluorescence test |
| CN110079450A (en) * | 2019-05-10 | 2019-08-02 | 广州安必平医药科技股份有限公司 | It is a kind of for the solid phase carrier chip system of in situ hybridization and its application |
| CN110218790A (en) * | 2019-05-10 | 2019-09-10 | 广州安必平医药科技股份有限公司 | For detecting probe chip, kit and its application of the rearrangement of Ph-like ALL related gene |
| CN113913500A (en) * | 2021-09-15 | 2022-01-11 | 武汉友芝友医疗科技股份有限公司 | Probe set for detecting ALK gene rearrangement and application thereof |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6022689A (en) * | 1995-04-07 | 2000-02-08 | University Of New Mexico | Situ hybridization slide processes |
| CN1417348A (en) * | 2001-11-09 | 2003-05-14 | 联合基因科技有限公司 | Gene chip for multiple-sample detection |
| CN1926246A (en) * | 2004-03-01 | 2007-03-07 | 仓敷纺绩株式会社 | Hybridization method as well as hybridization microarray and hybridization kit |
| CN102369295A (en) * | 2009-02-26 | 2012-03-07 | 丹麦达科有限公司 | Compositions and methods for performing hybridizations with separate denaturation of the sample and probe |
| US20140178875A1 (en) * | 2012-10-09 | 2014-06-26 | Advandx, Inc. | Devices, Compositions and Methods Pertaining To Microscopic Analysis Of Microorganisms and Other Analytes of Interest |
| CN207537461U (en) * | 2017-11-09 | 2018-06-26 | 武汉康录生物技术股份有限公司 | A kind of multiprobe hybridized slides |
-
2017
- 2017-11-09 CN CN201711095572.2A patent/CN107904283A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6022689A (en) * | 1995-04-07 | 2000-02-08 | University Of New Mexico | Situ hybridization slide processes |
| CN1417348A (en) * | 2001-11-09 | 2003-05-14 | 联合基因科技有限公司 | Gene chip for multiple-sample detection |
| CN1926246A (en) * | 2004-03-01 | 2007-03-07 | 仓敷纺绩株式会社 | Hybridization method as well as hybridization microarray and hybridization kit |
| CN102369295A (en) * | 2009-02-26 | 2012-03-07 | 丹麦达科有限公司 | Compositions and methods for performing hybridizations with separate denaturation of the sample and probe |
| US20140178875A1 (en) * | 2012-10-09 | 2014-06-26 | Advandx, Inc. | Devices, Compositions and Methods Pertaining To Microscopic Analysis Of Microorganisms and Other Analytes of Interest |
| CN207537461U (en) * | 2017-11-09 | 2018-06-26 | 武汉康录生物技术股份有限公司 | A kind of multiprobe hybridized slides |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108709991A (en) * | 2018-06-20 | 2018-10-26 | 浙江天杭生物科技股份有限公司 | A kind of slide and its application method of bovine viral Immunofluorescence test |
| CN110079450A (en) * | 2019-05-10 | 2019-08-02 | 广州安必平医药科技股份有限公司 | It is a kind of for the solid phase carrier chip system of in situ hybridization and its application |
| CN110218790A (en) * | 2019-05-10 | 2019-09-10 | 广州安必平医药科技股份有限公司 | For detecting probe chip, kit and its application of the rearrangement of Ph-like ALL related gene |
| CN113913500A (en) * | 2021-09-15 | 2022-01-11 | 武汉友芝友医疗科技股份有限公司 | Probe set for detecting ALK gene rearrangement and application thereof |
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