本發(fā)明涉及重排驅(qū)動(dòng)實(shí)體腫瘤融合基因檢測(cè)試劑、試劑盒及檢測(cè)系統(tǒng)，屬于生物醫(yī)藥。

背景技術(shù)：

1、基因融合(重排、易位)是腫瘤驅(qū)動(dòng)變異中一類(lèi)非常重要的形式。實(shí)體腫瘤中包括一類(lèi)融合基因驅(qū)動(dòng)的腫瘤，類(lèi)型包括軟組織腫瘤、子宮間葉源性腫瘤、涎腺腫瘤以及一些其他類(lèi)型的實(shí)體腫瘤等。融合基因的識(shí)別對(duì)于重排驅(qū)動(dòng)的實(shí)體腫瘤病理診斷、分型具有重要作用。舉例來(lái)說(shuō)，軟組織腫瘤來(lái)源廣泛，組織學(xué)表現(xiàn)各異，其年發(fā)病率2.4-5/10萬(wàn)。惡性軟組織腫瘤約占成人惡性腫瘤的1％，約占兒童惡性腫瘤15％，好發(fā)年齡為30-50歲。最常見(jiàn)的發(fā)病部位包括：四肢(60％)和軀干部(19％)。已知軟組織肉瘤有100多種亞型。軟組織肉瘤患者5年生存率為60％-80％。軟組織腫瘤的病理診斷是病理領(lǐng)域中的難點(diǎn)之一，一是其比較少見(jiàn)，使得很多病理醫(yī)師對(duì)軟組織腫瘤復(fù)雜的組織學(xué)形態(tài)不太熟悉，二是不同類(lèi)型的軟組織腫瘤在組織學(xué)上存在較大的重疊性，同時(shí)存在很多假肉瘤性病變和中間性腫瘤，使不少病理醫(yī)師在軟組織腫瘤的病理診斷上存在困難。某些肉瘤直接以基因重排進(jìn)行命名，如cic-nutm1基因融合肉瘤是一種cic基因融合肉瘤的新分子亞型，好發(fā)于中樞神經(jīng)系統(tǒng)，多見(jiàn)于兒童，該腫瘤的表型可能與nut癌混淆。子宮間葉源性的腫瘤和涎腺腫瘤也屬于較為罕見(jiàn)的腫瘤，在診斷上也較大程度需要依賴(lài)融合基因的檢出結(jié)果。除此之外，實(shí)體瘤中發(fā)生基因融合變異的比例約為16.5％-20％，其中約36％可作為分子治療靶點(diǎn)。針對(duì)alk、ros1、ret、fgfr2和ntrk1/2/3等融合靶點(diǎn)，fda已經(jīng)批準(zhǔn)了多款相應(yīng)的靶向藥物。以非小細(xì)胞肺癌為例，fda、nmpa已經(jīng)批準(zhǔn)了克唑替尼、塞瑞替尼、阿來(lái)替尼、brigatinib、lorlatinib和恩沙替尼等藥物，用于治療攜帶alk融合的nsclc患者，具有較高的反應(yīng)率和持久應(yīng)答，提高了患者的生存期并改善了生活質(zhì)量。因此，識(shí)別融合基因?qū)τ谠\斷、分型、治療都有非常重要的意義。

2、重排驅(qū)動(dòng)的實(shí)體腫瘤涉及的融合基因斷點(diǎn)多、分布廣、伴侶基因多。傳統(tǒng)檢測(cè)方法包括熒光原位雜交(fish)、免疫組化(ihc)、聚合酶鏈反應(yīng)(rt-pcr)等，是腫瘤融合基因檢測(cè)不可或缺的手段，但其分辨率和通量通常較低，只能檢測(cè)臨床意義非常明確的幾個(gè)基因的特定融合，不能識(shí)別新的融合基因伴侶或復(fù)雜的結(jié)構(gòu)重排。fish方法僅能針對(duì)特定基因進(jìn)行融合判定，且不能確定基因伴侶。免疫組化方法利用的是抗原抗體特異性結(jié)合的原理，可在蛋白水平對(duì)融合導(dǎo)致的高蛋白表達(dá)基因進(jìn)行檢測(cè)，該方法可檢測(cè)的蛋白有一定局限性。rt-pcr方法需要對(duì)發(fā)生融合的兩個(gè)基因分別設(shè)計(jì)引物，僅能檢測(cè)已知的融合形式。這些方法均屬于低通量檢測(cè)方法，一次檢測(cè)僅針對(duì)一種或幾種已知融合或融合伴侶形式，僅適用于在腫瘤組織診斷形態(tài)上有一定傾向性的情況時(shí)，對(duì)特定相應(yīng)的融合進(jìn)行確認(rèn)或排除。對(duì)于復(fù)雜的臨床病例，需要同時(shí)做多種標(biāo)志物來(lái)確定診斷，這對(duì)病理醫(yī)生水平有很高的要求，對(duì)樣本量也有很大要求，很多小腫瘤或穿刺病例將失去明確診斷機(jī)會(huì)。ngs(高通量測(cè)序/下一代測(cè)序)檢測(cè)方法用于融合基因的檢測(cè)，從建庫(kù)方法考慮包括擴(kuò)增法和雜交捕獲方法；從檢測(cè)材料考慮包括dna-based和rna-based檢測(cè)方法；從檢測(cè)范圍考慮包括靶向測(cè)序和全基因測(cè)序。ngs擴(kuò)增法同樣需要針對(duì)融合形式的兩個(gè)伴侶基因都已知的情況，根據(jù)兩個(gè)基因斷裂位點(diǎn)附近序列設(shè)計(jì)引物，因此，僅能檢測(cè)已知融合形式。雜交捕獲ngs方法可以在融合基因一端覆蓋探針的情況下即可檢出該變異，可以用于未知融合形式的檢出，適用于融合伴侶多的病例?；赿na水平的ngs常用于已知基因已知斷點(diǎn)的常見(jiàn)基因重排形式的檢測(cè)，因?yàn)槿诤匣驍嗔盐稽c(diǎn)通常位于內(nèi)含子，由于內(nèi)含子區(qū)域成本問(wèn)題，一般無(wú)法完全覆蓋。如果內(nèi)含子序列較長(zhǎng)，內(nèi)含子區(qū)域存在重復(fù)序列、特定內(nèi)含子低質(zhì)量重復(fù)和復(fù)雜重排都很難設(shè)計(jì)引物或探針。且當(dāng)融合變異豐度過(guò)低，剪接異常所致的融合在dna水平檢測(cè)時(shí)很有可能會(huì)漏檢。特別對(duì)于疑難病例中不確定的融合形式、融合伴侶無(wú)法檢出。國(guó)內(nèi)外多個(gè)隊(duì)列研究表明基于rna為模板的ngs(rna-ngs)檢測(cè)融合/重排、易位要顯著優(yōu)于dna為模板的ngs(dna-ngs)，基于rna層面進(jìn)行基因融合檢測(cè)可能更有意義。由于成本限制，靶向ngs方法可以設(shè)置檢測(cè)更高的深度，相比于全基因測(cè)序更適用于臨床使用。

3、本發(fā)明提供的靶向rna融合基因的檢測(cè)試劑在rna水平進(jìn)行融合基因檢測(cè)，采用雜交捕獲的高通量測(cè)序方法，在檢測(cè)能力方面更具有優(yōu)勢(shì)。對(duì)于實(shí)體腫瘤，包括軟組織腫瘤、涎腺腫瘤、子宮相關(guān)腫瘤以及其他融合基因驅(qū)動(dòng)的腫瘤的輔助診斷具有非常重要的意義；本發(fā)明可同時(shí)檢出多種基因融合形式，包括未知的融合伴侶形式，可提供具體融合形式和斷裂位點(diǎn)位置。為患者提供了精準(zhǔn)診斷治療機(jī)會(huì)，同時(shí)也避免了因多種標(biāo)志物疊加帶來(lái)的經(jīng)濟(jì)負(fù)擔(dān)和額外增加的風(fēng)險(xiǎn)。

技術(shù)實(shí)現(xiàn)思路

1、本發(fā)明的目的是為解決如何獲得一種重排驅(qū)動(dòng)實(shí)體腫瘤靶向rna融合基因的檢測(cè)試劑及檢測(cè)系統(tǒng)，以實(shí)現(xiàn)對(duì)實(shí)體腫瘤，包括軟組織腫瘤、涎腺腫瘤、子宮相關(guān)腫瘤以及其他融合基因驅(qū)動(dòng)的腫瘤的基因進(jìn)行檢測(cè)的技術(shù)問(wèn)題。

2、為達(dá)到解決上述問(wèn)題的目的，本發(fā)明提供一種用于檢測(cè)人體實(shí)體瘤融合基因的檢測(cè)探針庫(kù)，所述探針庫(kù)包括與下列一個(gè)或多個(gè)染色體位點(diǎn)雜交的探針；所述染色體位點(diǎn)共計(jì)11472處。

3、具體染色體位點(diǎn)如下：

4、chr1|246580618|246580738,chr1|71870146|71870266,chr1|71866666|71866786,chr1|6845464|6845584,chr1|9787787|9787907,chr1|37957304|37957424,chr1|37957544|37957664,chr1|37970744|37970864,chr1|179073235|179073355,chr1|179079806|179079926,chr1|115259887|115260007,chr1|115260127|115260247,chr1|115260247|115260367,chr1|71867626|71867746,chr1|115260007|115260127,chr1|115260367|115260487,chr1|9786947|9787067,chr1|37958504|37958624,chr1|71870266|71870386,chr1|115259527|115259647,chr1|115259647|115259767,chr1|115260487|115260607,chr1|115260727|115260847,chr1|120512234|120512354,chr1|156849841|156849961,chr1|164769026|164769146,chr1|164815801|164815921,chr1|179073355|179073475,chr1|243716131|243716251,chr1|6931837|6931957,chr1|6931957|6932077,chr1|7811249|7811369,chr1|9782242|9782362,chr1|9782566|9782686,chr1|9784007|9784127,chr1|9784274|9784394,chr1|9784394|9784514,chr1|9784822|9784942,chr1|9787067|9787187,chr1|9787187|9787307,chr1|9787307|9787427,chr1|9787427|9787547,chr1|9787547|9787667,chr1|9787667|9787787,chr1|9788147|9788267,chr1|9788267|9788387,chr1|9788387|9788507,chr1|9788507|9788627,chr1|9788627|9788747,chr1|9788747|9788867,chr1|9788867|9788987,chr1|37956104|37956224,chr1|37957904|37958024,chr1|37958984|37959104,chr1|37959104|37959224,chr1|37959224|37959344,chr1|37959344|37959464,chr1|37959464|37959584,chr1|37959584|37959704,chr1|37967365|37967485,chr1|37967485|37967605,chr1|37975026|37975146,chr1|37978974|37979094,chr1|65344728|65344848,chr1|65533197|65533317,chr1|71866906|71867026,chr1|115259767|115259887,chr1|115260607|115260727,chr1|115261184|115261304,chr1|115261304|115261424,chr1|115262250|115262370,chr1|115263150|115263270,chr1|120459206|120459326,chr1|120461069|120461189,chr1|120462005|120462125,chr1|120471629|120471749,chr1|120508026|120508146,chr1|120510790|120510910,chr1|120512114|120512234,chr1|120539600|120539720,chr1|120539720|120539840,chr1|120539840|120539960,chr1|120547852|120547972,chr1|120547972|120548092,chr1|120548092|120548212,chr1|120572499|120572619,chr1|120611918|120612038,chr1|120612038|120612158,chr1|120612158|120612278,chr1|164529131|164529251,chr1|164761841|164761961,chr1|164776749|164776869,chr1|164789319|164789439,chr1|164818441|164818561,chr1|164818561|164818681,chr1|164818681|164818801,chr1|164818801|164818921,chr1|164818921|164819041,chr1|164819041|164819161,chr1|164819161|164819281,chr1|164819281|164819401,chr1|164819401|164819521,chr1|164819521|164819641,chr1|164819641|164819761,chr1|164819761|164819881,chr1|164819881|164820001,chr1|164820001|164820121,chr1|164820121|164820241,chr1|164820241|164820361,chr1|164820361|164820481,chr1|164820481|164820601,chr1|164820601|164820721,chr1|179079686|179079806,chr1|179086457|179086577,chr1|179089305|179089425,chr1|179090770|179090890,chr1|179095582|179095702,chr1|179095702|179095822,chr1|243736218|243736338,chr1|243800933|243801053,chr1|245912609|245912729,chr1|245912729|245912849,chr1|245912849|245912969,chr1|6845584|6845704,chr1|6867033|6867153,chr1|6880231|6880351,chr1|6885062|6885182,chr1|6885182|6885302,chr1|6931717|6931837,chr1|6946290|6946410,chr1|6946410|6946530,chr1|6946530|6946650,chr1|6946650|6946770,chr1|6946770|6946890,chr1|6946890|6947010,chr1|6947010|6947130,chr1|6947130|6947250,chr1|6947250|6947370,chr1|6947370|6947490,chr1|6947490|6947610,chr1|6947610|6947730,chr1|6947730|6947850,chr1|6947850|6947970,chr1|6947970|6948090,chr1|6948090|6948210,chr1|6948210|6948330,chr1|7151334|7151454,chr1|7309501|7309621,chr1|7309621|7309741,chr1|7527880|7528000,chr1|7700390|7700510,chr1|7700510|7700630,chr1|7721726|7721846,chr1|7721846|7721966,chr1|7723393|7723513,chr1|7723513|7723633,chr1|7723633|7723753,chr1|7723753|7723873,chr1|7723873|7723993,chr1|7723993|7724113,chr1|7724113|7724233,chr1|7724233|7724353,chr1|7724353|7724473,chr1|7724473|7724593,chr1|7724593|7724713,chr1|7724713|7724833,chr1|7724833|7724953,chr1|7724953|7725073,chr1|7725073|7725193,chr1|7725193|7725313,chr1|7729760|7729880,chr1|7729880|7730000,chr1|7730881|7731001,chr1|7731001|7731121,chr1|7737629|7737749,chr1|7737749|7737869,chr1|7740068|7740188,chr1|7740188|7740308,chr1|7764756|7764876,chr1|7764876|7764996,chr1|7792468|7792588,chr1|7792588|7792708,chr1|7796394|7796514,chr1|7796514|7796634,chr1|7796981|7797101,chr1|7797295|7797415,chr1|7797415|7797535,chr1|7797535|7797655,chr1|7797979|7798099,chr1|7798099|7798219,chr1|7798219|7798339,chr1|7798339|7798459,chr1|7798459|7798579,chr1|7804865|7804985,chr1|7804985|7805105,chr1|7805815|7805935,chr1|7805935|7806055,chr1|7806055|7806175,chr1|7807750|7807870,chr1|7809821|7809941,chr1|7811369|7811489,chr1|7812509|7812629,chr1|7815648|7815768,chr1|7826499|7826619,chr1|7826619|7826739,chr1|7826739|7826859,chr1|7826859|7826979,chr1|7826979|7827099,chr1|7827099|7827219,chr1|7827219|7827339,chr1|7827339|7827459,chr1|7827459|7827579,chr1|7827579|7827699,chr1|7827699|7827819,chr1|7827819|7827939,chr1|7827939|7828059,chr1|7828059|7828179,chr1|7828179|7828299,chr1|7828299|7828419,chr1|7828419|7828539,chr1|7828539|7828659,chr1|782865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07|41573727,chr22|41573727|41573847,chr22|41573967|41574087,chr22|41574087|41574207,chr22|41574207|41574327,chr22|41574327|41574447,chr22|41574447|41574567,chr22|41574567|41574687,chr22|41574807|41574927,chr22|41574927|41575047,chr22|41575047|41575167,chr22|41575167|41575287,chr22|41575287|41575407,chr22|41575407|41575527,chr22|41575527|41575647,chr22|41575647|41575767,chr22|41575767|41575887,chr22|41576007|41576127,chrx|66765193|66765313,chrx|48115496|48115616,chrx|70352645|70352765,chrx|48126519|48126639,chrx|48126279|48126399,chrx|48126399|48126519,chrx|48126759|48126879,chrx|48126639|48126759,chrx|48123317|48123437,chrx|48125712|48125832,chrx|48121181|48121301,chrx|48123197|48123317,chrx|48117211|48117331,chrx|48117961|48118081,chrx|48117091|48117211,chrx|48116647|48116767,chrx|70361120|70361240,chrx|66765073|66765193,chrx|47030537|47030657,chrx|66915625|66915745,chrx|70360515|70360635,chrx|70360635|70360755,chrx|66915505|66915625,chrx|76761188|76761308,chrx|48114797|48114917,chrx|66944108|66944228,chrx|76761308|76761428,chrx|39911199|39911319,chrx|39911439|39911559,chrx|39911559|39911679,chrx|39913119|39913239,chrx|39913239|39913359,chrx|39914571|39914691,chrx|39914691|39914811,chrx|39921442|39921562,chrx|39922089|39922209,chrx|39922851|39922971,chrx|39922971|39923091,chrx|39923091|39923211,chrx|39923649|39923769,chrx|39930176|39930296,chrx|39930850|39930970,chrx|39931562|39931682,chrx|39931682|39931802,chrx|39931802|39931922,chrx|39932042|39932162,chrx|39932282|39932402,chrx|39932402|39932522,chrx|39932522|39932642,chrx|39932642|39932762,chrx|39932762|39932882,chrx|39933002|39933122,chrx|39933122|39933242,chrx|39933242|39933362,chrx|39933482|39933602,chrx|39933722|39933842,chrx|39933842|39933962,chrx|39933962|39934082,chrx|39934082|39934202,chrx|39934202|39934322,chrx|39935667|39935787,chrx|48888982|48889102,chrx|50051935|50052055,chrx|50053615|50053735,chrx|50053975|50054095,chrx|50054095|50054215,chrx|50054215|50054335,chrx|50054335|50054455,chrx|50055527|50055647,chrx|50056846|50056966,chrx|50085251|50085371,chrx|50089721|50089841,chrx|50090675|50090795,chrx|50094576|50094696,chrx|50094696|50094816,chrx|50094816|50094936,chrx|66766273|66766393,chrx|70354197|70354317,chrx|70356121|70356241,chrx|70357556|70357676,chrx|70357676|70357796,chrx|70362135|70362255,chrx|76761068|76761188,chrx|76761428|76761548,chrx|76761548|76761668,chrx|76761668|76761788,chrx|76761788|76761908,chrx|76761908|76762028,chrx|76762028|76762148,chrx|76762148|76762268,chrx|76762268|76762388,chrx|76762388|76762508,chrx|76762508|76762628,chrx|76762628|76762748,chrx|76762748|76762868,chrx|76762868|76762988,chrx|76762988|76763108,chrx|76907784|76907904,chrx|39910479|39910599,chrx|39910599|39910719,chrx|39910719|39910839,chrx|39910839|39910959,chrx|39910959|39911079,chrx|39911079|39911199,chrx|39911319|39911439,chrx|39913479|39913599,chrx|39916368|39916488,chrx|39916488|39916608,chrx|39921322|39921442,chrx|39921562|39921682,chrx|39921969|39922089,chrx|39922209|39922329,chrx|39923529|39923649,chrx|39923769|39923889,chrx|39930296|39930416,chrx|39931922|39932042,chrx|39932162|39932282,chrx|39932882|39933002,chrx|39933362|39933482,chrx|39933602|39933722,chrx|39934322|39934442,chrx|39936987|39937107,chrx|39937107|39937227,chrx|39956448|39956568,chrx|39956568|39956688,chrx|39956688|39956808,chrx|39956808|39956928,chrx|39956928|39957048,chrx|39957048|39957168,chrx|39957168|39957288,chrx|40036240|40036360,chrx|40036360|40036480,chrx|40036480|40036600,chrx|47004560|47004680,chrx|47004680|47004800,chrx|47004800|47004920,chrx|47006696|47006816,chrx|47006816|47006936,chrx|47028684|47028804,chrx|47028804|47028924,chrx|47030417|47030537,chrx|47032477|47032597,chrx|47034408|47034528,chrx|47035889|47036009,chrx|47038492|47038612,chrx|47038678|47038798,chrx|47038798|47038918,chrx|47039259|47039379,chrx|47039379|47039499,chrx|47039601|47039721,chrx|47039798|47039918,chrx|47040604|47040724,chrx|47040724|47040844,chrx|47040856|47040976,chrx|47040976|47041096,chrx|47041058|47041178,chrx|47041178|47041298,chrx|47041340|47041460,chrx|47041521|47041641,chrx|47041641|47041761,chrx|47044394|47044514,chrx|47044514|47044634,chrx|47044681|47044801,chrx|47044831|47044951,chrx|47044951|47045071,chrx|47045105|47045225,chrx|47045454|47045574,chrx|47045577|47045697,chrx|47045697|47045817,chrx|47045853|47045973,chrx|47045973|47046093,chrx|47046093|47046213,chrx|48886197|48886317,chrx|48886317|48886437,chrx|48886437|48886557,chrx|48886557|48886677,chrx|48886677|48886797,chrx|48886797|48886917,chrx|48886917|48887037,chrx|48887037|48887157,chrx|48887157|48887277,chrx|48887277|48887397,chrx|48887397|48887517,chrx|48887517|48887637,chrx|48887637|48887757,chrx|48887757|48887877,chrx|48887877|48887997,chrx|48887997|48888117,chrx|48888862|48888982,chrx|48890960|48891080,chrx|48891181|48891301,chrx|48891549|48891669,chrx|48891669|48891789,chrx|48895525|48895645,chrx|48895692|48895812,chrx|48895812|48895932,chrx|48895932|48896052,chrx|48896582|48896702,chrx|48896702|48896822,chrx|48896822|48896942,chrx|48897952|48898072,chrx|48898072|48898192,chrx|48900597|48900717,chrx|48900717|48900837,chrx|48900837|48900957,chrx|49969366|49969486,chrx|49969486|49969606,chrx|50027526|50027646,chrx|50028037|50028157,chrx|50028157|50028277,chrx|50031770|50031890,chrx|50037853|50037973,chrx|50037973|50038093,chrx|50051455|50051575,chrx|50051575|50051695,chrx|50051695|50051815,chrx|50051815|50051935,chrx|50052055|50052175,chrx|50052175|50052295,chrx|50052295|50052415,chrx|50052415|50052535,chrx|50052535|50052655,chrx|50052655|50052775,chrx|50052775|50052895,chrx|50052895|50053015,chrx|50053015|50053135,chrx|50053135|50053255,chrx|50053255|50053375,chrx|50053375|50053495,chrx|50053495|50053615,chrx|50053735|50053855,chrx|50053855|50053975,chrx|50054455|50054575,chrx|50085131|50085251,chrx|50089601|50089721,chrx|50090555|50090675,chrx|50094170|50094290,chrx|50094290|50094410,chrx|66763753|66763873,chrx|66763873|66763993,chrx|66763993|66764113,chrx|66764113|66764233,chrx|66764233|66764353,chrx|66764353|66764473,chrx|66764473|66764593,chrx|66764593|66764713,chrx|66764713|66764833,chrx|66764833|66764953,chrx|66764953|66765073,chrx|66765313|66765433,chrx|66765433|66765553,chrx|66765553|66765673,chrx|66765673|66765793,chrx|66765793|66765913,chrx|66765913|66766033,chrx|66766033|66766153,chrx|66766153|66766273,chrx|66766393|66766513,chrx|66766513|66766633,chrx|66788638|66788758,chrx|66788758|66788878,chrx|66863088|66863208,chrx|66863208|66863328,chrx|66905773|66905893,chrx|66905893|66906013,chrx|66909368|66909488,chrx|66909488|66909608,chrx|66914425|66914545,chrx|66914545|66914665,chrx|66914665|66914785,chrx|66914785|66914905,chrx|66914905|66915025,chrx|66915025|66915145,chrx|66915145|66915265,chrx|66915265|66915385,chrx|66915385|66915505,chrx|66915745|66915865,chrx|66915865|66915985,chrx|66931214|66931334,chrx|66931334|66931454,chrx|66931454|66931574,chrx|66937250|66937370,chrx|66937370|66937490,chrx|66941615|66941735,chrx|66941735|66941855,chrx|66942609|66942729,chrx|66942729|66942849,chrx|66943508|66943628,chrx|66943628|66943748,chrx|66943748|66943868,chrx|66943868|66943988,chrx|66943988|66944108,chrx|66944228|66944348,chrx|66944348|66944468,chrx|66944468|66944588,chrx|66944588|66944708,chrx|66944708|66944828,chrx|66944828|66944948,chrx|66944948|66945068,chrx|66945068|66945188,chrx|66945188|66945308,chrx|66945308|66945428,chrx|66945428|66945548,chrx|66945548|66945668,chrx|66945668|66945788,chrx|66945788|66945908,chrx|66945908|66946028,chrx|66946028|66946148,chrx|66946148|66946268,chrx|66946268|66946388,chrx|66946388|66946508,chrx|66946508|66946628,chrx|66946628|66946748,chrx|66946748|66946868,chrx|66946868|66946988,chrx|66946988|66947108,chrx|66947108|66947228,chrx|66947228|66947348,chrx|66947348|66947468,chrx|66947468|66947588,chrx|66947588|66947708,chrx|66947708|66947828,chrx|66947828|66947948,chrx|66947948|66948068,chrx|66948068|66948188,chrx|66948188|66948308,chrx|66948308|66948428,chrx|66948428|66948548,chrx|66948548|66948668,chrx|66948668|66948788,chrx|66948788|66948908,chrx|66948908|66949028,chrx|66949028|66949148,chrx|66949148|66949268,chrx|66949268|66949388,chrx|66949388|66949508,chrx|66949508|66949628,chrx|66949628|66949748,chrx|66949748|66949868,chrx|66949868|66949988,chrx|66949988|66950108,chrx|66950108|66950228,chrx|66950228|66950348,chrx|66950348|66950468,chrx|70338426|70338546,chrx|70338546|70338666,chrx|70338666|70338786,chrx|70339213|70339333,chrx|70339526|70339646,chrx|70339646|70339766,chrx|70339814|70339934,chrx|70339934|70340054,chrx|70340811|70340931,chrx|70340931|70341051,chrx|70341167|70341287,chrx|70341362|70341482,chrx|70341482|70341602,chrx|70341602|70341722,chrx|70342000|70342120,chrx|70342120|70342240,chrx|70342348|70342468,chrx|70342508|70342628,chrx|70342628|70342748,chrx|70342905|70343025,chrx|70343025|70343145,chrx|70343394|70343514,chrx|70343514|70343634,chrx|70343999|70344119,chrx|70344119|70344239,chrx|70344594|70344714,chrx|70344796|70344916,chrx|70344916|70345036,chrx|70345171|70345291,chrx|70345291|70345411,chrx|70345493|70345613,chrx|70345816|70345936,chrx|70345936|70346056,chrx|70346101|70346221,chrx|70346221|70346341,chrx|70346759|70346879,chrx|70346879|70346999,chrx|70347136|70347256,chrx|70347256|70347376,chrx|70347733|70347853,chrx|70347853|70347973,chrx|70348136|70348256,chrx|70348256|70348376,chrx|70348368|70348488,chrx|70348488|70348608,chrx|70348954|70349074,chrx|70349136|70349256,chrx|70349256|70349376,chrx|70349470|70349590,chrx|70349590|70349710,chrx|70349835|70349955,chrx|70349955|70350075,chrx|70351360|70351480,chrx|70351853|70351973,chrx|70351973|70352093,chrx|70352207|70352327,chrx|70352327|70352447,chrx|70352765|70352885,chrx|70352963|70353083,chrx|70354543|70354663,chrx|70354663|70354783,chrx|70354872|70354992,chrx|70354992|70355112,chrx|70356241|70356361,chrx|70356361|70356481,chrx|70356481|70356601,chrx|70356709|70356829,chrx|70356829|70356949,chrx|70357027|70357147,chrx|70357147|70357267,chrx|70357398|70357518,chrx|70360395|70360515,chrx|70361000|70361120,chrx|70361713|70361833,chrx|70362015|70362135,chrx|70362255|70362375,chrx|76760348|76760468,chrx|76760468|76760588,chrx|76760588|76760708,chrx|76760708|76760828,chrx|76760828|76760948,chrx|76760948|76761068,chrx|76763108|76763228,chrx|76763228|76763348,chrx|76763348|76763468,chrx|76763468|76763588,chrx|76763588|76763708,chrx|76763708|76763828,chrx|76763828|76763948,chrx|76763948|76764068,chrx|76764068|76764188,chrx|76776156|76776276,chrx|76776276|76776396,chrx|76776871|76776991,chrx|76777631|76777751,chrx|76777751|76777871,chrx|76778710|76778830,chrx|76778830|76778950,chrx|76812882|76813002,chrx|76813002|76813122,chrx|76814120|76814240,chrx|76814240|76814360,chrx|76829705|76829825,chrx|76845294|76845414,chrx|76849096|76849216,chrx|76849216|76849336,chrx|76854830|76854950,chrx|76854950|76855070,chrx|76855171|76855291,chrx|76855853|76855973,chrx|76855973|76856093,chrx|76872031|76872151,chrx|76872151|76872271,chrx|76874254|76874374,chrx|76874374|76874494,chrx|76875853|76875973,chrx|76875973|76876093,chrx|76888655|76888775,chrx|76888775|76888895,chrx|76888994|76889114,chrx|76889114|76889234,chrx|76890075|76890195,chrx|76891376|76891496,chrx|76891496|76891616,chrx|76907544|76907664,chrx|76907664|76907784,chrx|76909578|76909698,chrx|76912030|76912150,chrx|76918811|76918931,chrx|76918931|76919051,chrx|76920074|76920194,chrx|76920194|76920314,chrx|76931681|76931801,chrx|76937002|76937122,chrx|76937122|76937242,chrx|76937242|76937362,chrx|76937362|76937482,chrx|76937482|76937602,chrx|76937602|76937722,chrx|76937722|76937842,chrx|76937842|76937962,chrx|76937962|76938082,chrx|76938082|76938202,chrx|76938202|76938322,chrx|76938322|76938442,chrx|76938442|76938562,chrx|76938562|76938682,chrx|76938682|76938802,chrx|76938802|76938922,chrx|76938922|76939042,chrx|76939042|76939162,chrx|76939162|76939282,chrx|76939282|76939402,chrx|76939402|76939522,chrx|76939522|76939642,chrx|76939642|76939762,chrx|76939762|76939882,chrx|76939882|76940002,chrx|76940002|76940122,chrx|76940381|76940501,chrx|76944301|76944421,chrx|76949243|76949363,chrx|76949363|76949483,chrx|76952055|76952175,chrx|76952175|76952295,chrx|76953051|76953171,chrx|76954002|76954122,chrx|76972578|76972698,chrx|76972698|76972818,chrx|77041428|77041548,chrx|77041548|77041668,chrx|77041668|77041788,chrx|129114885|129115005,chrx|129115005|129115125,chrx|129115125|129115245,chrx|129115245|129115365,chrx|129115629|129115749,chrx|129116531|129116651,chrx|129116651|129116771,chrx|129116771|129116891,chrx|129139154|129139274,chrx|129139274|129139394,chrx|129146524|129146644,chrx|129146856|129146976,chrx|129146976|129147096,chrx|129147096|129147216,chrx|129147216|129147336,chrx|129147336|129147456,chrx|129147456|129147576,chrx|129147576|129147696,chrx|129147696|129147816,chrx|129147816|129147936,chrx|129147936|129148056,chrx|129148056|129148176,chrx|129148176|129148296,chrx|129148296|129148416,chrx|129148416|129148536,chrx|129148536|129148656,chrx|129148656|129148776,chrx|129148776|129148896,chrx|129148896|129149016,chrx|129149016|129149136,chrx|129149136|129149256,chrx|129149256|129149376,chrx|129149376|129149496,chrx|129149496|129149616,chrx|129149616|129149736,chrx|129149736|129149856,chrx|129149856|129149976,chrx|129149976|129150096,chrx|129150096|129150216,chrx|129154910|129155030,chrx|129155030|129155150,chrx|129156862|129156982,chrx|129158915|129159035,chrx|129159035|129159155,chrx|129159155|129159275,chrx|129159275|129159395,chrx|129162600|129162720,chrx|129162720|129162840,chrx|129168420|129168540,chrx|129168540|129168660,chrx|129171322|129171442,chrx|129171442|129171562,chrx|129173052|129173172,chrx|129173172|129173292,chrx|129184662|129184782,chrx|129185815|129185935,chrx|129185935|129186055,chrx|129189769|129189889,chrx|129189889|129190009,chrx|129190009|129190129,chrx|129190129|129190249,chrx|129190249|129190369,chrx|129190369|129190489,ch?rx|129190489|129190609,chrx|129190609|129190729,chrx|129190729|129190849,chrx|129190849|129190969,chrx|129190969|129191089,chrx|129191089|129191209,chrx|129191209|129191329,chrx|129191329|129191449,chrx|129191449|129191569,chrx|129191569|129191689,chrx|129191689|129191809,chr?x|129191809|129191929,chrx|129191929|129192049；

5、所述實(shí)體瘤包括軟組織腫瘤、子宮間葉源性腫瘤或涎腺腫瘤等重排驅(qū)動(dòng)的腫瘤。

6、本發(fā)明提供一種用于檢測(cè)人體實(shí)體瘤融合基因的檢測(cè)試劑，所述試劑包括用于檢測(cè)融合基因的探針組合，所述探針組合來(lái)自一探針庫(kù)，所述探針庫(kù)包括與上述一個(gè)或多個(gè)染色體位點(diǎn)雜交的探針。

7、本發(fā)明提供一種用于檢測(cè)人體實(shí)體瘤融合基因的檢測(cè)試劑盒，所述試劑盒包括用于檢測(cè)融合基因的探針組合，所述探針組合來(lái)自一探針庫(kù)，所述探針庫(kù)包括與上述一個(gè)或多個(gè)染色體位點(diǎn)雜交的探針

8、本發(fā)明提供根據(jù)上述的探針在制備用于檢測(cè)人體實(shí)體瘤融合基因的試劑或試劑盒中的應(yīng)用。

9、本發(fā)明提供一種人體實(shí)體瘤融合基因的檢測(cè)系統(tǒng)，包括：檢測(cè)模塊和分析模塊；所述檢測(cè)模塊可用于對(duì)待測(cè)的實(shí)體腫瘤樣本的rna進(jìn)行提取，逆轉(zhuǎn)錄，基因捕獲及文庫(kù)構(gòu)建，通過(guò)高通量測(cè)序，獲得測(cè)序結(jié)果；所述分析模塊，用于對(duì)高通量測(cè)序結(jié)果進(jìn)行計(jì)算機(jī)處理，分析獲得的融合基因的信息；所述檢測(cè)模塊中包括用于檢測(cè)融合基因的探針組合，所述探針組合來(lái)自一探針庫(kù)，所述探針庫(kù)包括與上述一個(gè)或多個(gè)染色體位點(diǎn)雜交的探針。

10、相比現(xiàn)有技術(shù)，本發(fā)明具有如下有益效果：

11、1、本發(fā)明提供的檢測(cè)試劑盒在基因選擇上結(jié)合了復(fù)旦大學(xué)附屬腫瘤醫(yī)院既往的研究結(jié)果，并囊括了臨床指南以及現(xiàn)有文獻(xiàn)中所報(bào)的融合相關(guān)基因，檢測(cè)更加全面，符合病理診斷需求，解決了實(shí)體腫瘤尤其是軟組織腫瘤、涎腺腫瘤、子宮相關(guān)腫瘤以及其他融合驅(qū)動(dòng)的腫瘤的診斷難的問(wèn)題。

12、2、本發(fā)明采用基于rna水平的雜交捕獲的高通量測(cè)序法?？赏瑫r(shí)檢測(cè)多基因相關(guān)融合形式，并提供檢測(cè)基因表達(dá)值，輔助融合基因檢測(cè)。所包含的融合基因伴侶即使是未知的或不在本發(fā)明檢測(cè)試劑所包含基因中的，也可被檢出。具有高靈敏度、高效的特點(diǎn)。

13、3、本發(fā)明提供的探針、試劑盒和方法是基于探針捕獲的rna-based?ngs檢測(cè)技術(shù)，覆蓋了目前已被報(bào)道的實(shí)體瘤融合基因；一次檢測(cè)即可以高效助力臨床腫瘤的精準(zhǔn)分型和診斷，不僅可以解決傳統(tǒng)檢測(cè)方法通量低的問(wèn)題，而且克服了dna-based?ngs檢測(cè)方法檢測(cè)融合時(shí)存在的挑戰(zhàn)(斷點(diǎn)多在內(nèi)含子上，且內(nèi)含子區(qū)域大，探針很難做到覆蓋所有內(nèi)含子區(qū)域，容易導(dǎo)致漏檢)，并且能夠檢出新型伴侶基因融合(多重pcr方法做不到)，顯著提高檢測(cè)融合變異的靈敏度和特異性。

14、4、本發(fā)明先對(duì)rna逆轉(zhuǎn)錄形成cdna，再采用dna探針進(jìn)行目標(biāo)基因捕獲。相較于rna探針，dna探針更穩(wěn)定，降解速率更低，結(jié)合效率更高，對(duì)于提高基因檢測(cè)的穩(wěn)定性和準(zhǔn)確性具有明顯的提升。

15、5、本發(fā)明經(jīng)復(fù)旦大學(xué)附屬腫瘤醫(yī)院病理科使用過(guò)程中多次技術(shù)迭代，覆蓋更全面，技術(shù)更完善，經(jīng)過(guò)嚴(yán)格性能驗(yàn)證和臨床檢驗(yàn)實(shí)踐，對(duì)于疾病的診斷、分型、和治療都提供了極大的幫助。