CN102424848B - Probe combination for diagnosing Xp11.2 translocation renal cancer and application thereof - Google Patents
Probe combination for diagnosing Xp11.2 translocation renal cancer and application thereof Download PDFInfo
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Abstract
The invention discloses a probe combination for diagnosing Xp11.2 translocation renal cancer and an application thereof, which belong to the field of fluorescent in situ hybridization probe application. The probe combination is BAC cloning fragments RP11-58H17, RP11-352D11, RP11-416B14 and RP11-107C19. The specificity and the sensibility of the Xp11.2 translocation renal cancer diagnosis by the probe combination respectively reach 100 percent, convenience, high speed and reliability are realized, in addition, the success ratio is high, and the probe combination can be used for preparing Xp11.2 translocation renal cancer diagnosis reagents.
Description
Technical field
The invention belongs to the fluorescence in situ hybridization probe Application Areas, relate to a kind of for the probe combinations of diagnosis Xp11.2 transposition kidney and in the application for preparing Xp11.2 transposition Diagnosis of Renal Cell Carcinoma reagent.
Background technology
WHO revised renal cell carcinoma histopathology classification in 1997 in 2004, had increased Xp11.2 transposition/TFE3 gene fusion dependency kidney (Xp11.2 transposition kidney) newly, this rare tumor types.Although its overall sickness rate is very low, about 1/3 is this type of kidney in the renal cell carcinoma in children patient, and retrospective study shows that this type of tumour is unrare in China.This project application people diagnosed and had reported 11 these tumours of example at home first with the method for immunohistochemical methods in 2007.Up-to-date studies show that, the prognosis of Xp11.2 transposition kidney is than non-Xp11.2 transposition kidney poor prognosis.And the prognosis of different genes type Xp11.2 transposition kidney is distinguished to some extent.In addition, there is positive evidence to show that the patient of Xp11.2 transposition kidney is to vascular endothelial growth factor receptor (vascular endothelial growth factor receptor, VEGFR) or Mammals rapamycin (mammalian target of rapamycin, mTOR) molecular targeted therapy responsive.Other there are some researches show that the MET Tyrosylprotein kinase is the target gene of ASPL-TFE3 fusion gene, and is expected to become the treatment target spot of Xp11.2 transposition kidney.Therefore according to genotype, come this type of tumour of Precise Diagnosis to seem very important.
The molecular pathology feature of this tumour is to exist to comprise Xp11.2 in interior chromosome translocation, finally all causes comprising the TFE3 transcription factor gene in interior gene fusion.Have at least at present 7 kinds of different transposition forms to be in the news, but clear and definite site only have 5 kinds, comprise t (X; 1) (p11.2; Q21), cause PRCC and TFE3 gene fusion; T (X; 17) (p11.2; Q25), cause ASPL and TFE3 gene fusion; T (X; 1) (p11.2; P34), cause PSF and TFE3 gene fusion; Inv (X) (p11; Q12), cause NonO (p54nrb) and TFE3 gene fusion; T (X; 17) (p11.2; Q23), cause the CLTC-TFE3 gene fusion.Think at present because the conversion of promotor the final high expression level TFE3 of these fusion genes fusion rotein.Therefore using immunohistochemical methods detection nucleus TFE3 fusion rotein is one of current diagnosis Xp11.2 transposition kidney important method.But its shortcoming is the method is subject to various factors easily, as organizes set time, tissue repair mode, antibody cloning number and artificial interpretation factor etc.So that false positive or false negative appear in the possibility of result.Especially diagnosis is often more difficult when Histological Study is not true to type.
In addition, find by the research to fusion gene mRNA, the node of all fusions of such tumour or transposition all is positioned at starting point or the terminal point of exon, so we can't know fusion and the transposition node of dna level, also just can not detect fusion gene with the method for PCR at dna level.Because this tumour is relatively rare, therefore difficult flesh tissue sample that obtains adopts the RT-PCR method to detect the TFE3 fusion gene, or judges that by the observation of cell caryogram diagnostic method of fusion gene type is actually rare in the work, and its operation is not convenient yet.More accurate and easily the molecular diagnosis method await further foundation.
Chromosome fluorescence in-situ hybridization (FISH) starts from traditional cytogenetics and the combination of dna technique, and rapid sensitive, specificity are good, can detect concealment or small chromosome aberration and complex karyotype; Can also use multiple fluorescent mark, show relative position and direction between dna fragmentation and the gene, spatial positioning is accurate.In addition, the method for FISH can be carried out retrospective study at paraffin-embedded sample, greatly reduces the requirement to the research sample.At present, utilize fluorescence in situ hybridization (FISH) to diagnose the external report of method of this tumour less, the domestic report that there is not yet.
Similar probe in the foreign literature:
[1]Argani?P，Aulmann?S，Karanjawala?Z，Fraser?RB，Ladanyi?M，Rodriguez?MM.Melanotic?Xp11?translocation?renal?cancers：a?distinctive?neoplasm?with?overlapping?features?of?PEComa，carcinoma，and?melanoma.Am?J?Surg?Pathol.2009?Apr；33(4)：609-619.
S?Aulmann，T?Longerich，P?Schirmacher，G?Mechtersheimer?&?R?Penzel?Detection?of?the?ASPSCR1-TFE3?gene?fusion?in?paraffin-embedded?alveolar?soft?part?sarcomasHistopathology?2007，50，881-886.
Used probe: RPCI11-211H10, RPCI11-58H17, RPCI11-122 N23, RPCI-57A11 (kinetochore end) RPCI11-552E4, RPCI11-344N17, RPCI11-735G22 (telomere end).
Estimate: this probe has used more cloned sequence somewhat expensive, and only verifies in 2 routine Xp11.2 transposition/TFE3 gene fusion dependency kidneys and 5 routine alveolar soft part sarcomas.
[2]Zhong?M，De?Angelo?P，Osborne?L，Keane-Tarchichi?M，Goldfischer?M，Edelmann?L，Yang?Y，Linehan?WM，Merino?MJ，Aisner?S，Hameed?M.Dual-color，break-apart?FISH?assay?on?paraffin-embedded?tissues?as?an?adjunct?to?diagnosis?of?Xp11?translocation?renal?cell?carcinoma?and?alveolar?soft?part?sarcoma.Am?J?Surg?Pathol.2010?Jun；34(6)：757-766.
Used probe: RP11-404P16 (458kb) and RP11-416B14 (182kb) telomere end RP11-528A24 (116kb) and RP11-58H17 (182kb) kinetochore end
Estimate: the cloned sequence of this probe is not of uniform size, and maximum and the kb more than 300 that differs minimum produce the heterogeneity of in situ hybridization condition easily.And only in 5 routine Xp11.2 transposition/TFE3 gene fusion dependency kidneys and 2 routine alveolar soft part sarcomas, verified.
Summary of the invention
The objective of the invention is provides a kind of probe combinations for diagnosis Xp11.2 transposition kidney for above-mentioned technical problem.
Another object of the present invention provides the application of above-mentioned probe combinations in preparation Xp11.2 transposition Diagnosis of Renal Cell Carcinoma reagent.
Further object of the present invention provides the diagnostic kit that contains above-mentioned probe combinations.
The objective of the invention is to realize by following technical proposal:
A kind of probe combinations for diagnosis Xp11.2 transposition kidney, this is combined as BAC clone segment (perhaps being BAC clone probe) RP11-58H17, RP11-352D11, RP11-416B14, RP11-107C19.
Described probe combinations, wherein RP11-58H17 and RP11-352D11 are positioned TFE3 kinetochore one side, and the two is labeled as the fluorescence of any one identical color; RP11-416B14 and RP11-107C19 are positioned TFE3 telomere one side, and the two is labeled as identical but different from the color of kinetochore one side mark fluorescence.
Described probe combinations, wherein RP11-58H17 and RP11-352D11 all are labeled as green fluorescence, and RP11-416B14 and RP11-107C19 all are labeled as red fluorescence; The fluorescence color of mark can exchange.
The application of described probe in preparation Xp11.2 transposition Diagnosis of Renal Cell Carcinoma reagent.
A kind of Xp11.2 transposition Diagnosis of Renal Cell Carcinoma test kit is characterized in that this test kit comprises probe claimed in claim 1.
Below be the detailed explanation of technical solution of the present invention:
The probe combinations that the present invention adopts is based on the analysis of the situations such as distributing position to the different BAC cloned sequence binding site at TFE3 gene two ends on the karyomit(e), size and the preferred version taked unlike the prior art.
Among the present invention, TFE3 kinetochore one side BAC cloned sequence is RP11-58H17 (fragment length 200kb) and RP11-352D11 (fragment length 175kb), TFE3 telomere one side BAC cloned sequence is RP11-416B14 (fragment length 182kb) and RP11-107C19 (fragment length 160kb), these fragments are bacterial artificial chromosome (Bacterial artificial chromosome, BAC) clone, its location on human chromosomal is open, is respectively RP11-58H17 (X chromosome 49444577-49644633) X chromosome, RP11-352D11 (X chromosome 49071535-49246795), RP11-416B14 (X chromosome 48465815-48648142), RP11-107C19 (X chromosome 48172092-48332445).The orientating as of TFE3 gene (X chromosome 48771185-48787934).The BAC cloned sequence sequentially is RP11-58H17, RP11-352D11, TFE3, RP11-416B14, RP11-107C19 (from kinetochore side direction telomere side direction) with linking of TFE3 gene.
Keep certain distance between the BAC cloned sequence of combination on TFE3 gene and the karyomit(e) and between the adjacent BAC cloned sequence and can overlapping (be 283kb among the present invention to the maximum, minimum is not 98kb).Like this, because the BAC cloned sequence size that adopts close (differ between minimum and maximum among the present invention and only be 40kb), maximum distance is controlled in the 1500kb between the BAC cloned sequence of two ends, there is each other suitable interval, so that when carrying out Fluirescence observation, two BAC cloned sequences of telomere side show a kind of fluorescence, such as redness, side two BAC cloned sequences in kinetochore show another kind of fluorescence, such as green, when non-Xp11.2 transposition kidney, the TFE3 gene does not rupture, red green two kinds of fluorescence lean on closerly, occur red green fusion signal (showing as red green linking to each other or the yellow signal point) during observation, and when Xp11.2 transposition kidney, the TFE3 gene break, cause red green two kinds of fluorescence distant from getting, need not during observation to amplify and to see separation red green two kinds of signals far away, be easy to observe.
The BAC cloned sequence that connects the fluorescence of 2 mark colors of the same race at the every end of X chromosome TFE3 gene is in order to strengthen fluorescence intensity and scope, observe by experiment, the fluorescence intensity of 2 BAC cloned sequences and scope are enough observed, both avoided 1 BAC cloned sequence the too small situation of the inadequate scope of fluorescence intensity may occur, and avoided again adopting a plurality of BAC cloned sequences can cause the fluorescence scope to disperse to produce easily disturbing and also higher drawback of cost.
Select these 4 kinds of BAC cloned sequences that the consideration of the following aspects is arranged: 1. these several BAC cloned sequence sizes are close, and the benefit of bringing is: the fluorescence intensity of each end is consistent, and prevent that the end from crossing strong and the other end is crossed weak and impact is observed; The in situ hybridization condition is kept consistency.2. binding site makes between the adjacent BAC cloned sequence and can keep suitable distance, can strengthen each end fluorescence intensity and scope.TFE3 gene two ends BAC cloned sequence maximum distance is controlled within the 1500kb, there is the TFE3 gene between two kinds of fluorescence colors, merge signal (as showing as red green linking to each other or the yellow signal point) so that when the TFE3 gene does not rupture, present, and two kinds of fluorescence color separation are distant when the TFE3 gene break, are easy to observe.Otherwise if TFE3 gene two ends BAC cloned sequence maximum distance is excessive, as surpassing 1500kb even larger, then no matter whether the TFE3 gene ruptures, and all observes two kinds of fluorescence of obvious separation, just is difficult to judge whether to exist Xp11.2 transposition kidney.
BAC cloned sequence mark fluorescent is convenient to adopt fluorescence microscope, and is convenient directly perceived.Certainly, the BAC cloned sequence also is not limited to mark fluorescent, can be marked as the used marker of corresponding detection technique when adopting other detection techniques.
Beneficial effect of the present invention:
The present invention is according to the characteristics of Xp11.2 transposition kidney, design is combined in the fluorescent label DNA probe combinations at TFE3 gene two ends, in situ hybridization is carried out on basis in the paraffin-embedded tissue section, detects and merges and separation signal, can greatly improve the accuracy rate of such tumour of diagnosis.For molecular targeted therapy provides foundation.According to our experimental result, specificity and the susceptibility of the diagnosis of this probe combinations have all reached 100%, and operand only need to carry out in the paraffin-embedded tissue section, and the time only is two working dayss.Adopt probe combinations provided by the invention to detect Xp11.2 transposition kidney, not only easily and fast, reliable but also success ratio is high, can be used for preparing Xp11.2 transposition Diagnosis of Renal Cell Carcinoma test kit, for the fast and accurately diagnosis of Xp11.2 transposition kidney provides new instrument.
Description of drawings
Fig. 1: BAC clone probe positioning mode chart.
Fig. 2: the detection figure of 3 routine Xp11.2 transposition kidney fusion genes.
Wherein: figure A, figure B RT-PCR result show that 3 routine tumor tissues and cancer beside organism all detect an about 190-bp ASPL-TFE3 fusion gene band of size (figure A) and the about 200-bpTEF3-ASPL fusion gene band of another size (figure B); There are the balance transposition in figure C No. 17 karyomit(e)s of sequencing result demonstration and X chromosome, and ASPL-TFE3 fusion gene size is 195-bp (the ASPL exon 7 links to each other with TFE3 exon 4).TEF3-ASPL fusion gene size is 218-bp (the TFE3 exon 3 links to each other with ASPL exon 8).
Fig. 3: the orthographic plan of organization chip.
Fig. 4: visible 2 red green fusion signals in each nucleus of normal female nephridial tissue (the red green lap of arrow indication shows as yellow, observes real in leaning on to get very near red and green signals after amplifying), it is all complete to represent 2 X chromosomes.
Fig. 5: visible 1 red green fusion signal in each nucleus of normal male nephridial tissue (the red green lap of arrow indication shows as yellow, observes real in leaning on to get very near red and green signals after amplifying), it is complete to represent 1 X chromosome.
Fig. 6: interior visible 1 the red green fusion signal of each nucleus in male sex's Papillary Renal Cell Carcinoma (the red green lap of arrow indication shows as yellow, and observing real after amplifying is to lean on to get very near red and green signals), the not green separation signal of show.
Fig. 7: interior visible 1 the red green fusion signal of each nucleus in male sex's clear cell carcinoma of kidney (the red green lap of arrow indication shows as yellow, and observing real after amplifying is to lean on to get very near red and green signals), the not green separation signal of show.
Fig. 8: visible 1 pair of red green separation signal in (male sex) Xp11.2 transposition kidney tumor tissue cell nuclear (need not to amplify can clearly observe red and green signals separate far away) represents an X chromosome TFE3 gene break.
Fig. 9: (red green lap shows as yellow to visible 1 the red green fusion signal of arrow indication in (women) Xp11.2 transposition kidney tumor tissue cell nuclear, observe real in leaning on to get very near red and green signals after amplifying) and 1 pair of red green separation signal (need not amplification can clearly observe red and green signals separate far away), represent complete and another X chromosome TFE3 gene break of X chromosome.
Embodiment
The invention will be further elaborated by the following examples.
Probe described in the embodiment is the BAC cloned sequence, also can be the BAC clone probe.
The preparation of embodiment 1:DNA probe combinations:
2 BAC cloned sequences that selection can connect respectively at X chromosome TFE3 gene two ends, maximum distance keeps certain distance not overlapping in 1500kb between the probe of control two ends between the BAC cloned sequence, and clip size is close.The cloned sequence source is the human BAC clone center (http://www.empiregenomics.com/helixhq/clonecentral/search/human) of EmpireGenomics company.TFE3 kinetochore one side BAC clone segment is RP11-58H17 (200kb) and RP11-352D11 (175kb), and TFE3 telomere one side BAC clone segment is RP11-416B14 (182kb) and RP11-107C19 (160kb).The BAC cloned sequence sequentially is RP11-58H17, RP11-352D11, TFE3, RP11-416B14, RP11-107C19 with linking of TFE3 gene.The locating structure of probe combinations as shown in Figure 1.Utilize the nick translation method two BAC cloned sequences of telomere side to be marked as the fluorescence of any one identical color, preferred red fluorescence, two BAC cloned sequences of kinetochore side are marked as identical but different from the color of kinetochore one side mark fluorescence, preferred green fluorescence; The fluorescence color of two ends mark can exchange.The method is well known to those skilled in the art (the BAC cloned sequence of mark fluorescent is provided by U.S. EmpireGenomics company).TFE3 kinetochore two BAC cloned sequences of one side are a green fluorescence signal under fluorescent microscope, represent TFE3 gene kinetochore side.Two BAC cloned sequences of TFE3 telomere one side are a red fluorescence signal under fluorescent microscope, represent TFE3 gene telomere side.The fluorescence color of two ends mark can exchange.Red and green signals links to each other or is overlapping for merging signal under normal circumstances, when Xp11.2 transposition kidney, because X chromosome TFE3 gene break transposition is so that the red and green signals separation.
Further, the probe combinations of preparation can be adopted fluorescence in-situ hybridization method, whether its location of checking and/or diagnosis effect be reliable in Xp11.2 transposition kidney, non-Xp11.2 transposition kidney and cancer beside organism.
Embodiment 2: the fluorescence in situ hybridization process:
One, the organization chip of sample makes up:
Collect Nanjing General Hospital, Nanjing Military Area Command, PLA's Diagnosis of renal cell carcinoma 51 examples.By two veteran pathologist with reference to WHO 2004 uropoiesis and male reproductive system criteria for classification, immunohistochemical methods TFE3 is positive, and RT-PCR detects fusion gene result (Fig. 2 A, B, C, it is to have obtained diagnosis in the mRNA aspect that 3 examples are arranged among the 51 routine patients, with the mutual corresponding reliability that more can embody this experiment of this experimental result), re-start diagnostic assessment, last diagnostic Xp11.2 transposition kidney 24 examples as a result, clear cell carcinoma 9 examples, corpora mammillaria kidney 17 examples (sporadic corpora mammillaria kidney 16 examples wherein, the sick dependency corpora mammillaria of VHL kidney 1 example), unclassified renal cell carcinoma 1 example.Above sample (comprising cancer and cancer beside organism) is made into organization chip (Fig. 3).
Two, fluorescence in situ hybridization:
The thick section of organization chip 3 μ m after dewaxing, places 100%, 85%, 70% each 2min of ethanol successively, immerses subsequently in the deionized water 100 ℃ of water-bath 15min.Tissue slice is put into 37 ℃ of stomach en-K solution (0.1g stomach en-, 40ml 0.01M HCL), 15min; 2 * SSC (sodium-chlor, Trisodium Citrate) rinsing 2 times, each 5min, section places 0.1mol/L HCl soaking at room temperature 10min, with 2 * SSC rinsing 2 times, each 5min; Through 70%, 85%, the 100% ethanol 2min that respectively dewaters, in air drying; Mixed solution (each the 0.5 μ l of each probe wherein that adds the above-mentioned fluorescently-labeled probe of 10ul at tissue regions, 4 probes are totally 2 μ l, other adds the hybridization buffer of 8 μ l, hybridization buffer is provided by EmpireGenomics company when buying fluorescence labeling probe, include human Cot1 DNA), add cover glass, use the rubber adhesive edge; Put into 37 ℃ spend the night (16h) behind 88 ℃ of sex change 6min of in situ hybridization instrument (GeneAmp In Situ PCR System 1000); Remove cover glass, slide is placed 0.4 * SSC (sodium-chlor, Trisodium Citrate, 0.3%NP-40) solution, 69 ℃ of rinsing 1min; Rinsing 1min in 2 * SSC (sodium-chlor, Trisodium Citrate, 0.1%NP-40) solution, 70% ethanol 3min, the room temperature dark place is dry; Target region drips 10 μ l 4 ', 6-diamidino-2-phenylindone (4 ', 6-diamidino-2-phenylindole DAPI), after the cover glass mounting, use again the fluorescence microscope result.
The result judges:
The normal cell X chromosome, visible 1 the red green fusion signal of male sex's sample, visible 2 the red green fusion signals of women show as red green linking to each other or the yellow signal point.
Tumour cell, the abnormal signal of the visible a pair of red green separation of male sex's sample, visible normal signal and a pair of unusual red green separation signal of merging of women.
For getting rid of false positive and false negative, 100 cells of each sample counting, only have when 4 fluorescent signals (women) or 2 fluorescent signals (male sex) when all existing, just include the counting object in (in the women, we all can see 4 single signals (two green and two red) normal and tumour, be to merge in pairs at the normal female red and green signals only, look that being two merges signals, but actually formed by four single signals.See one and merge signal and a pair of red green single signal that separates in tumour, be 3 signals outwardly, but actual or be comprised of 4 single signals).Distance between the red green fluorescence signal is counted separation signal during greater than a deration of signal.When abnormal signal is designated as the positive greater than 10% the time.The judgment criteria that above as a result determination methods is exercised according to most similarly commercialization probes on the market is such as the probe using method of Vysis company and Dako company.
The result:
We detect 51 routine kidneys, wherein Xp11.2 transposition kidney 24 examples, clear cell carcinoma 9 examples, corpora mammillaria kidney 17 examples (wherein sporadic corpora mammillaria kidney 16 examples, the sick dependency corpora mammillaria of VHL kidney 1 example), unclassified renal cell carcinoma 1 example.24 routine Xp11.2 transposition kidneys all detect unusual separation signal as a result, (fluorescent microscope can't provide the visual field of 100 cells and observe for the people its positive cell number scope at 20%-85%, therefore the data of 20%-85% can only be examined repeatedly to count and get), the other normal kidney tissue of all the other tumours and cancer does not all detect abnormal signal (Fig. 2-9 has provided the representative positive picture of organization chip, the male sex and women Xp11.2 transposition kidney, all the other tumours such as the negative picture of clear cell carcinoma and corpora mammillaria kidney, the negative picture of men and women's normal kidney tissue).Illustrate and use specificity and the susceptibility of this probe in detecting Xp11.2 transposition kidney to be all 100%
Susceptibility=true positives/all Xp11.2 transposition kidney neoplastic disease number * 100%; 24 true positives case/24Xp11.2 transposition kidney=100%;
Specificity=true negative/all non-Xp11.2 transposition kidney neoplastic disease number * 100%; The non-Xp11.2 transposition in 27 true negative case/27 kidney tumour patient.
Estimate:
The cloned sequence that this group probe uses is relatively less, and the cloned sequence size is relatively consistent, differs between minimum and maximum only to be 40kb.Considered namely that in design economy considered again the consistence of in situ hybridization condition.In addition, the specificity of the double-colored break apart fluorescence in situ hybridization probe of TFE3 and susceptibility have all reached 100% in this experiment.Utilize the FISH technology, diagnose Xp11.2 transposition kidney with the double-colored break apart fluorescence in situ hybridization probe of TFE3.Fast, reliable and success ratio is high, be a new technology of diagnosis Xp11.2 transposition kidney, be worthy to be popularized.
Embodiment 3:Xp11.2 transposition Diagnosis of Renal Cell Carcinoma test kit
Contain embodiment 1 described probe combinations in the test kit, the feature of this probe combinations mainly is:
(1) TFE3 kinetochore one side BAC clone probe is RP11-58H17 (fragment length 200kb) and RP11-352D11 (fragment length 175kb), the mark green fluorescence.These two probes are a green fluorescence signal under fluorescent microscope, represent TFE3 gene kinetochore side.
(2) TFE3 telomere one side BAC clone probe is RP11-416B14 (fragment length 182kb) and RP11-107C19 (fragment length 160kb), the mark red fluorescence.These two probes are a red fluorescence signal under fluorescent microscope, represent TFE3 gene telomere side.
(3) red and green signals links to each other or is overlapping for merging signal under normal circumstances, when Xp11.2 transposition kidney, because X chromosome TFE3 gene break transposition is so that the red and green signals separation.
Claims (4)
1. a probe groups that is used for diagnosis Xp11.2 transposition kidney is characterized in that this probe groups is comprised of BAC clone segment RP11-58H17, RP11-352D11, RP11-416B14 and RP11-107C19;
Wherein: RP11-58H17 and RP11-352D11 are positioned TFE3 kinetochore one side, and the two is labeled as the fluorescence of any one identical color; RP11-416B14 and RP11-107C19 are positioned TFE3 telomere one side, and the two is labeled as identical but different from the color of kinetochore one side mark fluorescence.
2. probe groups according to claim 1 is characterized in that RP11-58H17 and RP11-352D11 all are labeled as green fluorescence, and RP11-416B14 and RP11-107C19 all are labeled as red fluorescence.
3. the application of probe groups claimed in claim 1 in preparation Xp11.2 transposition Diagnosis of Renal Cell Carcinoma reagent.
4. an Xp11.2 transposition Diagnosis of Renal Cell Carcinoma test kit is characterized in that this test kit comprises probe groups claimed in claim 1.
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