CN105483256A - BCR gene and ABL gene detection probe, preparation method thereof and reagent kit - Google Patents
BCR gene and ABL gene detection probe, preparation method thereof and reagent kit Download PDFInfo
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Abstract
The invention relates to a BCR gene and ABL gene detection probe and a preparation method thereof. The method includes the following steps that a selected BAC clone for a BCR gene is at least one of RP11-1026A5, RP11-165G5 and CTD-2079I4, a selected BAC clone is at least one of CTD-2302P22, CTD-2037J11 and CTD-2509L4, a selected BAC clone for an ABL is at least one of CTD-2037L19, RP11-21G10 and CTD-2526G20, the DNA of a plasmid is obtained, and marking is conducted. The invention further discloses a reagent kit provided with the BCR gene and ABL gene detection probe. By obtaining the optimal BCR gene and ABL detection probe through screening, signal line counting is accurate and quick, and result repeatability is good.
Description
Technical field
The invention belongs to biotechnology, particularly relate to BCR gene and abl gene detection probes and preparation method thereof and test kit.
Background technology
Chronic myelocytic leukemia (chronicmyelogenousleukemia, CML) is a kind of Clonal bone marrow proliferative tumour originating from hemopoietic stem cell, mainly involves granulocyte series, shows as lasting, Progressive symmetric erythrokeratodermia peripheral white blood cell amount increase.CML onset is slow, and initial symptoms is not obvious.Its natural history is that to be in progress by chronic phase be acceleration period, and finally develop into acute transformation phase, after the sudden turn of events, patient is everlasting dead in 3 ~ 5 months.
Constant, distinctive Ph karyomit(e) and molecular marker BCR/ABL fusion gene thereof is had in more than 90% Patient leukemic cell.The Ph karyomit(e) overwhelming majority is t (9; 22) (q34; Q11) typical transposition, small number of patients has variant traslocation, comprises No. 22 karyomit(e)s and the non-No. 9 simple variant traslocation of interchromosomal, 3 or more the interchromosomal complex translocations of bar (cryptic translocation).Ph karyomit(e) is present in the whole course of disease of CML, and after treatment is alleviated, still sustainable existence, therefore adopts bone marrow transplantation, eliminates Ph positive colony, just can reach final healing.
Therefore, by carrying out BCR/ABL detection to first visit patient, can CML be diagnosed, carrying out the examination of tyrosine kinase inhibitor beneficiaries.
Fluorescence in situ hybridization (FluorescenceinsituhybridizationFISH) is a kind of nonradioactivein situhybridization technology grown up on the basis of original radioactive in situ hybridization technology phase late 1980s.This technology current has been widely used in the structural research of animal-plant gene group, the analysis of variance of karyomit(e) fine structure, viral infection assays, mankind's antenatal diagnosis, cancer genetics and genome evolution research and has treated many fields.The ultimate principle of FISH is probe with known labeling nucleic acid, according to the principle of base complementrity, carries out anisogamy with single-chain nucleic acid unknown in material to be checked, form the heteroduplex nucleic acid that can be detected.Because DNA molecular is linearly arrange along the karyomit(e) longitudinal axis on chromosome, thus can probe directly and karyomit(e) carry out hybridizing thus specific gene located on chromosome.Compared with traditional radio-labeling in situ hybridization, fluorescence in situ hybridization has fast, detection signal is strong, hybrid specificities is high and can the feature such as multiple staining, is therefore subject to common concern in molecular cytogenetics field.
Hybridize probe used roughly can to classify three classes: 1) the special repetitive probe of karyomit(e), the such as probe of α satellite, satellite III class, its hybridization target position is often greater than 1Mb, not containing interspersed repeat sequence, be combined closely with target position, hybridization signal is strong, is easy to detect; 2) whole chromosome or chromosomal region specific probe, it is made up of extremely different nucleotide fragments on section a certain on item chromosome or karyomit(e), special large fragment can be obtained by the karyomit(e) be cloned in phage and plasmid; 3) specificity position probe, is made up of one or several cloned sequence.
The fluorescein-labelled of probe can adopt directly and the method for indirect labelling.Indirect labelling adopts biotin-labelled DNA probe, fluorescein avidin is associated with lotus root or Streptavidin detects after hybridization, Avidin-Biotin-luciferin complexes can also be utilized simultaneously, fluorescent signal be amplified, thus the fragment of about 500bp can be detected.And direct labelling method is by fluorescein directly and probe nucleotide or phosphopentose skeleton covalent attachment, or when nick-translation label probe, fluorescein ribonucleoside triphosphote is mixed.Direct labelling method step when detecting is simple, and Clinical practice is convenient.
And at present BCR/ABL gene FISH is detected, also lack the detection kit that specificity is high.
Summary of the invention
An object of the present invention is to provide a kind of BCR gene and abl gene detection probes and preparation method thereof, prepared probe can be used for detecting BCR gene and abl gene state, namely BCR gene and abl gene detection is detected, the fused type detected comprises typical transposition, variant traslocation and cryptic translocation, has good specificity.
The technical scheme realizing above-mentioned purpose is as follows.
A preparation method for BCR gene and abl gene detection probes, comprises the following steps:
(1) the BAC clone chosen for BCR gene is at least one in RP11-1026A5, RP11-165G5, CTD-2079I4, and chooses BAC clone at least one in CTD-2302P22, CTD-2037J11, CTD-2509L4; Clone as at least one in CTD-2037L19, RP11-21G10, CTD-2526G20 with choosing for the BAC of abl gene;
(2) respectively plasmid is extracted to clone, obtain plasmid DNA, quantitatively;
(3) fluorescein-labelled plasmid DNA is used, the color of the fluorescein marked with the detection probes for abl gene for BCR gene is not identical, and in BCR gene, to clone the color of the fluorescein of the probe of CTD-2302P22, CTD-2037J11, CTD-2509L4 not identical with deriving from BAC to derive from probe that BAC clones RP11-1026A5, RP11-165G5, CTD-2079I4, to obtain final product.
Wherein in an embodiment, the described BAC clone for abl gene is CTD-2037L19, RP11-21G10 and CTD-2526G20.
Wherein in an embodiment, the BAC clone for BCR gene is RP11-1026A5, RP11-165G5 and CTD-2079I4, and BAC clone is CTD-2302P22, CTD-2037J11 and CTD-2509L4.
Wherein in an embodiment, mark fluorescent element selects fluorescence dye known in the art, and preferably, fluorescein is Alexa
fITC, Alexa
rhodamine, TexasRed, pacific
dEAC.
Wherein in an embodiment, the mark of gene probe can adopt method of the prior art by corresponding fluorescein-labelled on double-strandednucleic acid, described method includes but not limited to: random priming, nick translation etc., mark gene probe can use commercially available nick translation labelling kit and random primer labelling kit, the NickTranslationKit of preferred abbott and Roche company.Step of the present invention (3) preferably adopts random priming, nick-translation method to carry out fluorescein-labelled to plasmid DNA.
Wherein in an embodiment, the temperature of described mark is 15 DEG C-18 DEG C, and the time of mark is 8-12 hour.
Another object of the present invention is to provide a kind of BCR gene and abl gene detection kit.
Realize this object technical scheme as follows.
A kind of BCR gene and abl gene detection kit, include above-mentioned BCR gene and abl gene detection probes.
Wherein in an embodiment, also include the COTHumanDNA for closed tumor-necrosis factor glycoproteins, and DAPI counterstain.
The present invention has following beneficial effect:
1. the present invention is by screening optimum BCR gene and abl gene detection probes and combination thereof, adopts FISH (FluorescenceIn-SituHybridization) method to realize detecting BCR/ABL fusion gene that is known or UNKNOWN TYPE; The qualification to BCR fracture mode can be realized.
2. carry out signal-count and result is reproducible accurately and rapidly by detection probes of the present invention; By FISH method to BCR/ABL fusion detection, supplement the deficiency of clinical middle BCR/ABL fusion detection, be conducive to accurately making a definite diagnosis CML, improve survival and Overall survival.
3. preferably to clone detection specificity good, highly sensitive in the present invention.By the visual inspection of resetting large fragment, not easily omit complicated variation type, also have good distinctive to unknown fused type.
4. by BCR gene of the present invention and ABL test kit, realize the application in the field such as oncobiology, cytogenetics, help each molecular marker of comprehensive evaluation, the medication of adjuvant clinical targeted therapy and treatment plan are selected.
Accompanying drawing explanation
Figure 1A is be the schematic diagram of abl gene detection probes sequence in embodiment 1.
Figure 1B is be the schematic diagram of BCR gene test probe sequence in embodiment 1.
Fig. 2 is human peripheral culturing cell sheet BCR gene and abl gene detection probes FISH detected result figure in embodiment 1.
Fig. 3 is clinical Bone Marrow sample FISH detected result figure in embodiment 4, and result display 2R2GA, has no BCR/ABL and merge.
Fig. 4 is clinical Bone Marrow sample FISH detected result figure in embodiment 4, and result display 1R1GA/1RG1RA, finds that BCR/ABL merges.
Fig. 5 is clinical Bone Marrow sample FISH detected result figure in embodiment 4, result display 1R1GA/1RG1RGA, BCR/ABL gene fusion, and is minor disruption type.
Embodiment
For the ease of understanding the present invention, will be described more fully the present invention below.The present invention can realize in many different forms, is not limited to embodiment described herein.On the contrary, provide the object of these embodiments be make the understanding of disclosure of the present invention more comprehensively thorough.
The experimental technique of unreceipted actual conditions in the following example, usual conveniently condition, the people such as such as Sambrook, molecular cloning: laboratory manual (NewYork:ColdSpringHarborLaboratoryPress, 1989) condition described in, or according to the condition that manufacturer advises.Various conventional chemical reagent used in embodiment, is commercially available prod.
Unless otherwise defined, all technology used in the present invention and scientific terminology are identical with belonging to the implication that those skilled in the art of the present invention understand usually.The object of the term used in specification sheets of the present invention just in order to describe specific embodiment, is not used in restriction the present invention.Term "and/or" used in the present invention comprises arbitrary and all combinations of one or more relevant Listed Items.
The preparation of embodiment 1BCR gene and abl gene detection probes
The preparation method of described ABL detection probes (GSPABL), comprises the following steps:
Colony screening: select the clone comprising goal gene ABL and two terminal sequences, as shown in Figure 1A.
GSPABL1 comprises the first probe, the second probe, the 3rd probe, four point probe, the 5th probe, can be one of them, several, or multiple combination.
The present embodiment selects the combination of 5 kinds of probes, table specific as follows, and it is bought in InvitrogenRP11BAC and CTDBAC clone bank.
ABL1 gene chr9:133,589,268-133,763,062,173,795bp
| Probe groups 1 | BAC | Insert Fragment start-stop position 3--> |
| First probe | CTD-2037L19 | chr9:133263327…133469440(206Kb) |
| Second probe | RP11-21G10 | chr9:133463380…133642562(179Kb) |
| 3rd probe | CTD-2526G20 | chr9:133,641,402-133,714,138(73Kb) |
| Four point probe | RP11-818C20 | chr9:133,733,184-133,922,526(189Kb) |
| 5th probe | RP11-728O24 | chr9:133,899,288-134,059,021(160Kb) |
The preparation method of described BCR detection probes (GSPBCR), comprises the following steps:
Colony screening: the clone selecting two terminal sequences comprised outside goal gene BCRM-BCR site prepares GSPBCR probe, as shown in Figure 1B.GSPBCR comprises two groups of probes, and these two groups of probes lay respectively at 3` end and the 5` end of BCR main faults point.
BCR gene chr22:23,522,552-23,660,224,137,673bp
| Probe groups 2 (left: green) | BAC | Insert Fragment start-stop position |
| 6th probe | RP11-1026A5 | chr22：23384725…23567903(183Kb) |
| 7th probe | RP11-165G5 | chr22：23222620…23362028(139Kb) |
| 8th probe | CTD-2079I4 | chr22：23249861…23407965(158Kb) |
(2) GSPBCR and GSPABL gene test probe preparation: the PlasmidMaxiKit using Qiagen company, the working method required to specifications carries out ultralow copy extraction of plasmid DNA respectively to different B AC clone, quantitative to plasmid DNA by the absorbancy measuring 260nm and 280nm place; Adopt autoclaved ultrapure water to dilute for 200ng/ul, adopt the centrifuge tube packing of 1.5ml, finally by the plasmid DNA mixing of 3 kinds or 5 kinds respective combination of the corresponding BCR gene that obtains or abl gene ,-20 DEG C of sealings are preserved.
(3) by nick translation method, fluorescent mark is carried out to plasmid DNA cocktail, fluorescein for group 2 probe mark of GSPBCR gene is green-dUTP, organize the fluorescein Aqua-dUTP of 3 probe marks, the fluorescein of often planting probe mark for GSPABL gene is red-dUTP.Adopt the NickTranslationKit of abbott, by following scheme, under strict lucifuge condition, prepare PCR reaction system on ice.
Join rear concussion mixing, mark 10 hours at 16 DEG C, then 80 DEG C hatch 10 minutes inactivators.Getting 5ul uses 2% sepharose to do electrophoresis, there is the band of disperse respectively at about 50-500bp.
Alcohol settling carried out to marked product and concentrate, in 1.5ml centrifuge tube, adding sodium-acetate and dehydrated alcohol by following scheme successively, lucifuge, on ice preparation:
Marked product 45ul
Sodium-acetate (3mol/L) 5ul
Dehydrated alcohol 125ul
Mixing to be placed in-70 DEG C of refrigerators at least 2 hours, and centrifugal 30 minutes of 4 DEG C of 13000rpm, carefully remove supernatant, do not stir precipitation, add 70% ethanol of 1ml, 4 DEG C 13000 revs/min centrifugal 15 minutes, carefully removes supernatant, do not stir precipitation, and lucifuge is dry.Use 1ul purified water dissolution precipitation, obtain GSPBCR and GSPABL gene probe, lucifuge ,-20 DEG C of storages.
4.GSPBCR and GSPABL gene probe is verified: the hybridization solution using ABL probe groups 1+BCR probe groups 2+BCR probe groups 3 to prepare respectively, uses mankind's proper splitting lymphocyte in mid-term to drip sheet and carries out probe checking (detection method reference example 3).Culturing cell comprises mid-term or interphase chromosome DNA, and during fluorescence in situ hybridization, chromosomal DNA shows as discernible karyomit(e) or nucleus in form.As shown in Fig. 2 (being applied to the experimental result of probe groups 1+ probe groups 2+ probe groups 3): the FISH results of hybridization figure of Metaphase Chromosome.In figure, visible BCR/ABL fusion gene detection probes signal becomes clear, and can be observed sensitivity, specificity 100% in human peripheral culturing cell sheet on Metaphase Chromosome; Use sample of bone marrow sheet to carry out hybridization check, clearly can record BCR and AB and close gene signal situation.Danger signal represents GSPABL1, and green and cyan signal represent GSPBCR main faults point two ends probe respectively.
Embodiment 2:BCR gene and abl gene detection reagent box preparation method
BCR gene and abl gene detection kit include BCR/ABL hybridization solution and DAPI counterstain two components, and wherein BCR and ABL hybridization solution comprises the combination of one group of GSPABL gene probe described in embodiment 1 and two groups of GSPBCR gene probes.BCR gene has two groups of detection probes, abl gene has one group of detection probes, in the present embodiment, described BCR gene and abl gene detection kit, for: BCR (group 2+ group 3)+ABL (group 1) combination, redye for the cell after the buffer composition of hybridizing environment (promoting hybridization), the DAPI counterstain such as COTHumanDNA of closing tumor-necrosis factor glycoproteins are mainly used in hybridization, DAPI wherein can be combined with DNA, make nucleus demonstrate blue-fluorescence, the counterstain containing Ursol D can keep the stable of fluorescence.
Concrete formula is as follows:
(1) hybridization solution preparation
(2) DAPI counterstain preparation
The Ursol D of 10mg is dissolved in the PBS of 1ml, regulates pH to be 9.0, adds 9ml glycerine, repeatedly shake mixing ,-20 DEG C of storages.The DAPI solution (0.1mg/ml) getting 2.5 μ l is dissolved in that 1ml is anti-to fade in liquid, repeatedly shakes mixing ,-20 DEG C of airtight preservations of lucifuge under lucifuge condition.
(3) finished product assembling
| Ingredient names | Specification/10test | Quantity |
| Hybridization solution | 100 μ l/ manage | 1 pipe |
| DAPI counterstain | 100 μ l/ manage | 1 pipe |
| Specification sheets | 1 part |
The detection method of embodiment 3:BCR gene and abl gene detection kit
1, sample preparation
1.1 get peripheral blood to be detected or the centrifugal 5min of marrow 2 ~ 3ml (heparin sodium anti-freezing) 2000rpm, carefully remove supernatant.
1.2 hypotonic mediums (0.075mol/LKCl) adding 10ml, piping and druming mixing, leaves standstill 3min.
1.337 ± 1 DEG C of hypotonic 30min of water bath.
1.4 add Fresh fixative 1ml, and piping and druming mixing, room temperature pre-fixes 10min.
1.5 piping and druming mixings, the centrifugal 5min of 2000rpm.
1.6 remove supernatant, and precipitation adds Fresh fixative 5 ~ 10ml, and piping and druming mixing, room temperature leaves standstill 10min.
1.72000rpm centrifugal 5min, removes supernatant.
1.8 can repeat above washing step, until cell precipitation whitens wash clean (this step does not need room temperature to leave standstill 10min).
2, film-making
2.1 get a clean slide glass;
Getting 3 μ l suspensions after 2.2 re-suspended cells is added drop-wise on slide glass;
2.3 dry under room temperature;
2.4, by 10 × object lens observation of cell density under phase microscope, require cell zero lap, and the wild cell quantity of haplopia is advisable at 100 ~ 200.
3, slide pre-treatment
The slide dripped is placed in room temperature 2 × SSC (PH7.0) solution by 3.1 soaks 2min;
3.2 successively in room temperature 70%, soaks 2min dehydration in the ethanol of 90%, 100%; Then take out slide, room temperature is dried.
4, sample and the same time variation of probe
4.1 add the hybridization solution of 10 μ l to hybridising region, cover rapidly 18 × 18mm cover glass, and light pressure makes hybridization solution be uniformly distributed, and avoid producing bubble;
4.2 use rubber glue along cover glass edge mounting, cover the position that cover glass contacts with slide glass completely;
Slide is put into hybridization instrument by 4.3, moistening in situ hybridization instrument humidity bar, inserts wet bar, covers hybridization instrument upper cover, arranges " Denat & Hyb " program, sex change 78 DEG C 2 minutes, hybridize 37 DEG C 10 ~ 18 hours.
5, post-hybridization washing and redying
5.1 slides put into 72 ± 1 DEG C of washing lotion I (1 × SSC/0.3%NP-40) 2 minutes;
5.2 take out slides, then to put it in room temperature washing lotion II (0.1%NP-40/2 × SSC) 30 seconds;
5.3 take out slides, then put it into room temperature 70%, 90%, in 100% ethanol each 2 minutes;
5.4 take out slide, dark place seasoning slide;
5.5 drip 10 μ lDAPI counterstains, to be seen.
6, interpretation of result
Fluorescence associated and DAPI need observe with suitable filter block.Count 50 ~ 200 cells, occur that the cell that RG merges, RA merges, RGA merges signal is designated as abnormal cells; Occur that the cell of 2R2GA signal type is designated as normal cell.
Embodiment 4:BCR gene and abl gene detection kit Clinical practice are evaluated
Use BCR gene described in embodiment 1 and the combination of abl gene detection probes, the combination of detection kit described in embodiment 2 (ABL (group 1)+BCR (group 2)+BCR (group 3), to 20 parts of clinical samples, (it is made a definite diagnosis through pathology detection, specifically see the following form), detect respectively.In triplicate, detected result is identical in the experiment of two kinds of probe combinations, detects consistence good.With commercial goods reagents ratio comparatively, detected result is completely the same, the specificity of reagent and highly sensitive.Fig. 3 and Fig. 4 is composite reagent box detected result, and shown in Fig. 3, signal type shows as 1R2GA, compares with normal signal 2R2GA, occurs that R signal lacks, so result is judged as that abl gene lacks, BCR/ABL gene fusion does not occur; Shown in Fig. 4, signal type shows as 1R1GA/1RG1RA, compares with normal signal 2R2GA, has occurred that GA signal ruptures, and R signal ruptures, and there occurs GR, AR signal fused simultaneously, so result is judged as BCR/ABL gene fusion, and is main faults type.Shown in Fig. 5, signal type shows as 1R1GA/1RG1RGA, compares with normal signal 2R2GA, has occurred G-signal fracture, R signal ruptures, and there occurs GR, GAR signal fused simultaneously, so result is judged as BCR/ABL gene fusion, and it is minor disruption type.
In the present invention, for abl gene and BCR gene, three groups of probes are used to achieve the detection of breaking point, fused type respectively.But probe combinations uses relatively, the use of combination clone probe, detection signal can be better.Probe length is longer in theory, and the fluorescent signal brightness obtained during actual detection is brighter, but because may relate to more polygene sequence, the signal complicacy possibility obtained increases, and also strengthens detecting the difficulty realized.The BAC of the detection probes of the probe groups 3 for the probe groups 1 of ABL1 gene, the probe groups 2 of BCR gene and BCR gene of the present invention clones its length and is respectively: 796Kb, 318Kb and 495Kb.
Contriver finds in probe checking of the present invention, and longer detection probes obtains stronger fluorescent signal really, and also obtain identical result in the detection validation of clinical sample.Therefore, in the design of fluorescent probe, luminance signals can be increased by proper extension fluorescent probe length, but specifically how to combinationally use, the certain technical difficulty existed, realize good detected result, except the experience in design, also need by clinical sample checking assessing signal type difference.
Know from above-mentioned laboratory test results, after molecular marker detection is carried out to these samples, molecule parting can be carried out according to detected result to sample, according to the meaning of Testing index, for clinical treatment formulation, medication selection and Outcome measure.
Each technical characteristic of described embodiment can combine arbitrarily, for making description succinct, all possible combination of each technical characteristic in above-described embodiment is not all described, but, as long as the combination of these technical characteristics does not exist contradiction, be all considered to be the scope that this specification sheets is recorded.
The above embodiment only have expressed several embodiment of the present invention, and it describes comparatively concrete and detailed, but can not therefore be construed as limiting the scope of the patent.It should be pointed out that for the person of ordinary skill of the art, without departing from the inventive concept of the premise, can also make some distortion and improvement, these all belong to protection scope of the present invention.Therefore, the protection domain of patent of the present invention should be as the criterion with claims.
Claims (8)
1. a preparation method for BCR gene and abl gene detection probes, is characterized in that, comprises the following steps:
(1) the BAC clone chosen for BCR gene is at least one in RP11-1026A5, RP11-165G5, CTD-2079I4, and chooses BAC clone at least one in CTD-2302P22, CTD-2037J11, CTD-2509L4; Clone as at least one in CTD-2037L19, RP11-21G10, CTD-2526G20 with choosing for the BAC of abl gene;
(2) respectively plasmid is extracted to clone, obtain plasmid DNA, quantitatively;
(3) fluorescein-labelled plasmid DNA is used, the color of the fluorescein marked with the plasmid DNA for abl gene for BCR gene is not identical, and in BCR gene, to clone the color of the fluorescein of the plasmid DNA of CTD-2302P22, CTD-2037J11, CTD-2509L4 not identical with deriving from BAC to derive from plasmid DNA that BAC clones RP11-1026A5, RP11-165G5, CTD-2079I4, to obtain final product.
2. the preparation method of BCR gene according to claim 1 and abl gene detection probes, is characterized in that, the described BAC clone for abl gene is CTD-2037L19, RP11-21G10 and CTD-2526G20.
3. the preparation method of BCR gene according to claim 1 and abl gene detection probes, it is characterized in that, BAC clone for BCR gene is RP11-1026A5, RP11-165G5 and CTD-2079I4, and is CTD-2302P22, CTD-2037J11 and CTD-2509L4 for BAC clones.
4. the preparation method of BCR gene according to claim 1 and abl gene detection probes, is characterized in that, described fluorescein is
fITC,
rhodamine, TexasRed,
or DEAC.
5. the preparation method of BCR gene and abl gene detection probes according to any one of claim 1-4, it is characterized in that, step (3) adopts random priming or nick-translation method to carry out fluorescein-labelled to plasmid DNA, the temperature of described mark is 15 DEG C-18 DEG C, and the time of mark is 8-12 hour.
6. the BCR gene that the preparation method according to any one of claim 1-5 obtains and abl gene detection probes.
7. BCR gene and an abl gene detection kit, is characterized in that, includes BCR gene according to claim 6 and abl gene detection probes.
8. BCR gene according to claim 7 and abl gene detection kit, is characterized in that, also includes the COTHumanDNA for closed tumor-necrosis factor glycoproteins, and DAPI counterstain.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201511033447.XA CN105483256A (en) | 2015-12-30 | 2015-12-30 | BCR gene and ABL gene detection probe, preparation method thereof and reagent kit |
| PCT/CN2016/105715 WO2017114008A1 (en) | 2015-12-30 | 2016-11-14 | Bcr gene and abl gene detection probe, preparation method therefor, and reagent kit |
Applications Claiming Priority (1)
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| WO2017114008A1 (en) * | 2015-12-30 | 2017-07-06 | 广州安必平医药科技股份有限公司 | Bcr gene and abl gene detection probe, preparation method therefor, and reagent kit |
| 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 |
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| US20050266459A1 (en) * | 2004-05-04 | 2005-12-01 | Poulsen Tim S | Nucleic acid probes and nucleic acid analog probes |
| CN103409505A (en) * | 2013-06-26 | 2013-11-27 | 武汉康录生物技术有限公司 | FISH (fluorescence in situ hybridization) probe, kit and detection method for detecting BCR/ABL fusion gene free from repetitive sequence |
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| CN101168772A (en) * | 2007-11-09 | 2008-04-30 | 冯文莉 | BCR/ABL fusion gene mRNA fluorescence quantitative PCR detecting kit |
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| WO2011140187A2 (en) * | 2010-05-04 | 2011-11-10 | University Of Rochester | Detecting chromosomal rearrangement |
| CN103667457B (en) * | 2012-04-25 | 2016-07-06 | 武汉艾迪康医学检验所有限公司 | The primer of detection leukemia BCR/ABL b3a2, b2a2 fusion gene relative expression quantity and method |
| CN105483256A (en) * | 2015-12-30 | 2016-04-13 | 广州安必平医药科技股份有限公司 | BCR gene and ABL gene detection probe, preparation method thereof and reagent kit |
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| US20050266459A1 (en) * | 2004-05-04 | 2005-12-01 | Poulsen Tim S | Nucleic acid probes and nucleic acid analog probes |
| CN103409505A (en) * | 2013-06-26 | 2013-11-27 | 武汉康录生物技术有限公司 | FISH (fluorescence in situ hybridization) probe, kit and detection method for detecting BCR/ABL fusion gene free from repetitive sequence |
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| WO2017114008A1 (en) * | 2015-12-30 | 2017-07-06 | 广州安必平医药科技股份有限公司 | Bcr gene and abl gene detection probe, preparation method therefor, and reagent kit |
| 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 |
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