腫瘤分子病理診斷基礎(chǔ)2

1、腫瘤分子病理診斷基礎(chǔ)閆慶國閆慶國分子病理與分子病理診斷分子病理與分子病理診斷腫瘤的分子病理變化腫瘤的分子病理變化腫瘤的分子病理診斷技術(shù)腫瘤的分子病理診斷技術(shù)腫瘤分子病理診斷的應(yīng)用腫瘤分子病理診斷的應(yīng)用腫瘤的分子病理診斷技術(shù)腫瘤的分子病理診斷技術(shù)Molecular pathology protocalsMolecular pathology protocalslDNA DNA extraction from paraffin-embedded tissuesextraction from paraffin-embedded tissueslDNA extraction from fresh or

2、 frozen tissues DNA extraction from fresh or frozen tissues lRNARNA extraction from fresh or frozen tissues extraction from fresh or frozen tissueslSingle-strand conformation polymorphism Single-strand conformation polymorphism analysis of analysis of mutations in exons 4-8 of the TP53 genemutations

3、 in exons 4-8 of the TP53 genelCleavaes fragment length polymorphism analysis for Cleavaes fragment length polymorphism analysis for genotyping and mutation detectiongenotyping and mutation detectionlDetection of telomerase by Detection of telomerase by in situ hybridization in situ hybridization an

4、d by and by the polymerase chain reaction based telomerase activity the polymerase chain reaction based telomerase activity assayassaylDetection of Detection of microsatellite instabilitymicrosatellite instabilitylPolymerase chain reaction Polymerase chain reaction clonality assays clonality assays

5、based on X-based on X-linked geneslinked geneslFluorescent in situ hybridizationFluorescent in situ hybridization: evaluation for ploidy and : evaluation for ploidy and gene amplificationgene amplificationlHer-2/neu oncogene Her-2/neu oncogene amplificationamplification determined by fluorescence de

6、termined by fluorescence in situ hybridizationin situ hybridizationlA nested A nested reversed transcription-polymerase chain reaction reversed transcription-polymerase chain reaction assay to detect BCR/ab/assay to detect BCR/ab/lDetection of t(15;17)(q24;q21),inv(16)/t(16;16)(p13;q22), Detection o

7、f t(15;17)(q24;q21),inv(16)/t(16;16)(p13;q22), and t(8;21)(q22;q22) anomalies in acute myeloid leukemiaand t(8;21)(q22;q22) anomalies in acute myeloid leukemialDetection of t(14;18)(q32;q21)-Associated BCL-2/JH Gene Detection of t(14;18)(q32;q21)-Associated BCL-2/JH Gene Fusion in Non-Hodgkins Lymph

8、omaFusion in Non-Hodgkins LymphomalDetection of Breast Cancer Cells Using Immunomagnetic Beads Detection of Breast Cancer Cells Using Immunomagnetic Beads and Reverse Transcription-Polymerase Chain Reactionand Reverse Transcription-Polymerase Chain ReactionlMolecular Detection of Molecular Detection

9、 of Circulating Prostate Cancer CellsCirculating Prostate Cancer CellslMethods to Detect Methods to Detect Clonal Gene Rearrangements Clonal Gene Rearrangements in Lymphomas in Lymphomas and Leukemiasand LeukemiaslMonitoring of Bone Marrow Transplant EngraftmentMonitoring of Bone Marrow Transplant E

10、ngraftmentlDirect Molecular Diagnosis of Multiple Endocrine Neoplasia Type Direct Molecular Diagnosis of Multiple Endocrine Neoplasia Type 1 1lMolecular Detection of Multiple Endocrine Neoplasia Type 2Molecular Detection of Multiple Endocrine Neoplasia Type 2lAssay for Detecting the I1307K Susceptib

11、ility Allele within the Assay for Detecting the I1307K Susceptibility Allele within the Adenomatous Polyposis Coli GeneAdenomatous Polyposis Coli GenelDetection of Detection of Human Papillomaviruses Human Papillomaviruses by Polymerase Chain by Polymerase Chain Reaction and In Situ HybridizationRea

12、ction and In Situ HybridizationlMolecular Methods for Detecting Molecular Methods for Detecting Epstein-Barr Virus Epstein-Barr Virus (Part I): (Part I): Hybridization to Epstein-Barr VirusEncoded RNA (EBER) Hybridization to Epstein-Barr VirusEncoded RNA (EBER) TranscriptsTranscriptslMolecular Metho

13、ds for Detecting Epstein-Barr Virus (Part II): Molecular Methods for Detecting Epstein-Barr Virus (Part II): Structural Analysis of Epstein-Barr Virus DNA as a Marker of Structural Analysis of Epstein-Barr Virus DNA as a Marker of ClonalityClonalitylMolecular Methods for Detecting Epstein-Barr Virus

14、 (Part III): Molecular Methods for Detecting Epstein-Barr Virus (Part III): EBV Viral Load by Competitive Polymerase Chain ReactionEBV Viral Load by Competitive Polymerase Chain ReactionlMolecular Detection of Kaposis SarcomaAssociated Molecular Detection of Kaposis SarcomaAssociated Herpesvirus/Hum

15、an Herpesvirus-8Herpesvirus/Human Herpesvirus-8lDiagnostic Applications of Quantitative Polymerase Chain Diagnostic Applications of Quantitative Polymerase Chain Reaction for Reaction for CytomegalovirusCytomegalovirus, Polymerase Chain Reaction , Polymerase Chain Reaction System That Detects System

16、 That Detects Herpes Simplex Virus Herpes Simplex Virus in Cerebrospinal in Cerebrospinal Fluid and Discriminates Genotypes 1 and 2Fluid and Discriminates Genotypes 1 and 2lDetection and Typing of Detection and Typing of Hepatitis CHepatitis C Virus ViruslDetection and Speciation of Detection and Sp

17、eciation of Mycobacteria Mycobacteria in Formalin-in Formalin-Fixed, Paraffin-Embedded Tissue SectionsFixed, Paraffin-Embedded Tissue SectionslUltrasensitive Quantitation of Human Immunodeficiency Ultrasensitive Quantitation of Human Immunodeficiency Virus Type 1 RNA in Plasma by the AMPLICOR and CO

18、BAS Virus Type 1 RNA in Plasma by the AMPLICOR and COBAS AMPLICOR AMPLICOR HIV-1HIV-1 MONITORTM Tests MONITORTM TestslMolecular Diagnosis of Molecular Diagnosis of HereditaryHereditary Thrombotic Disorders Thrombotic DisorderslPrenatal Genotyping Prenatal Genotyping of the RhD Locus to Identify of t

19、he RhD Locus to Identify Fetuses at Risk for Hemolytic Disease of the NewbornFetuses at Risk for Hemolytic Disease of the NewbornlMolecular Diagnosis of Hereditary HemochromatosisMolecular Diagnosis of Hereditary HemochromatosislGenotyping Genotyping of Apolipoprotein Eof Apolipoprotein ElGenotyping

20、 Genotyping for Functionally Important Human CYP2D6for Functionally Important Human CYP2D6* *4 4 (B) Mutation Using TaqMan Probes(B) Mutation Using TaqMan Probes常用的共性技術(shù)常用的共性技術(shù)u 核酸提取技術(shù)核酸提取技術(shù)u 電泳技術(shù)電泳技術(shù) u 凝膠成像凝膠成像u PCR PCR 技術(shù)技術(shù) 分子病理診斷的代表性技術(shù)分子病理診斷的代表性技術(shù) ISH In situ hybridizasionISH In situ hybridizasion

21、 FISHFISH Gene mutation analysisGene mutation analysis Clonality analysisClonality analysis FISH: Method FISH: Method Metaphase FISHMetaphase FISHPerformed on metaphase chromosome preparation.Performed on metaphase chromosome preparation.Advantage: Excellent resolution.Advantage: Excellent resolutio

22、n.Disadvantage: Collect live cells and culture in vitroDisadvantage: Collect live cells and culture in vitro. . Interphase FISHInterphase FISHPerformed on isolated nucleus or whole section.Performed on isolated nucleus or whole section.Advantage: Excellent correlation with histological Advantage: Ex

23、cellent correlation with histological and immunophenotypic features.Multiple probes on the and immunophenotypic features.Multiple probes on the same section.same section.Disadvantage: Nucleus and signal overlap truncation. Disadvantage: Nucleus and signal overlap truncation. FISH: Protocols for inte

24、rphase FISHFISH: Protocols for interphase FISHSlides preparationPrepare 2-4 m sections and dewax;Digest in 0.1% pepsin in 0.015N HCl for 20 min at 37C;Dehydrate and air dry slides.Hybridisation Apply probe to area of interest, place coverslip and seal with rubber cement;Denature probe and target DNA

25、 at 80C for 25 min;Hybridise in a moisture chamber at 45C for 2 days.Post-hybridisation washing Remove rubber cement and cover slip; Wash 1- 0.4x SSC and 0.3% IGEPAL CA-630 buffer in a water bath set at 72 C; Wash 2- 2SSC and 0.1% IGEPAL CA-630 buffer at room temp for 1 min;Wash 3 - 2SSC for 2-5 min

26、 at room temp;Apply anti-fade solution containing DAPI and coverslip.Read signals with a fluorescent microscope.FISH: ProbesDual-colour break-apart probe independent of translocation partnersNormal cell14 1414q32t(14;18)/IGH-BCL2 positive cell14 der (14)der (18) 18FISH: ProbesDual colour-dual fusion

27、 probe - translocation partners knownNormal cell14q3214 1418q211818t(14;18)/IGH-BCL2 positive cell14 der (14)der (18) 18 FISH: Probes Single colour probe CEP 8Chromosome specific to detect aneuplody17q11.2-q12 Her-2/neuCEP 17locus specific to detect copy number changes of a particular region (deleti

28、ons, gain or amplifications) Examples of specimen types applicable Examples of specimen types applicable to FISHto FISHFresh/frozen tissueCytology specimensBody fluids(urine)Intraoperative smearsCell culture preparationsFresh-frozen, parafin-embedded tissueThin sections(46)Disaggregated nucleiArchiv

29、ed unstained sectionsPreviously stained sections(e.g., negative immunohistochemistry controls)CISHDetection of gene amplification, chromosome translocations and chromosome number Using conventional enzymatic reactions under the brightfield microscope on formalin-fixed, paraffin-embedded tissuesCompa

30、red to FISH, CISHs advantages Detect under bright field The morphological details are readily apparent The probe signals are not subject to rapid fading2122突變檢測原理：突變檢測原理：根據(jù)野生型與突變型所表現(xiàn)出的差異根據(jù)野生型與突變型所表現(xiàn)出的差異 測序法：測序法：經(jīng)典、熒光、焦磷酸經(jīng)典、熒光、焦磷酸 （直接從序列上看差異）（直接從序列上看差異）定量定量 PCRPCR：探針法：探針法- -染料法染料法 ARMS-TaqMan ARMS-Ta

31、qMan （引物結(jié)合后反應(yīng)的差異（引物結(jié)合后反應(yīng)的差異- -間接）間接） HRM HRM、PNA-LNA PNA-LNA （根據(jù)產(chǎn)物變性溶解過程的差異（根據(jù)產(chǎn)物變性溶解過程的差異- -間接）間接）DHPLC DHPLC （根據(jù)不同構(gòu)象分子色譜分析的移動性差異（根據(jù)不同構(gòu)象分子色譜分析的移動性差異- -間接）間接）PCR-RFLP PCR-RFLP （酶切后電泳看泳動性差異（酶切后電泳看泳動性差異- -間接）間接） 直接測序法直接測序法焦磷酸測序焦磷酸測序根據(jù)已知序列，每合成一個(gè)核苷酸加一次樣，測一次熒光強(qiáng)度根據(jù)已知序列，每合成一個(gè)核苷酸加一次樣，測一次熒光強(qiáng)度150200E SG CA A G

32、 T G C T G AT G CT C G TT/TG/G51015焦磷酸測序焦磷酸測序 EGFR Exon 21 EGFR Exon 21 第第858858密碼子發(fā)生密碼子發(fā)生CTGCTG到到CGGCGG的突變，即發(fā)生了的突變，即發(fā)生了L858RL858R突變突變 150200E ST G C G T G T C A A C T A C GT/GT/T51015150EST GCG T G T C A A CT A C GT/TT/T51015ARMSARMS amplification refractory mutation system高分辨率熔解曲線分析高分辨率熔解曲線分析 High

33、 Resolution Melting, High Resolution Melting, HRMHRMPCRPCR產(chǎn)物中的突變位點(diǎn)改變產(chǎn)物中的突變位點(diǎn)改變了了DNADNA熔解曲線的形狀；純?nèi)劢馇€的形狀；純合子和雜合子樣本可以通過合子和雜合子樣本可以通過使用一種飽和型的使用一種飽和型的DNADNA結(jié)合結(jié)合染料輕松的區(qū)分開來，并顯染料輕松的區(qū)分開來，并顯示出清晰的熔解曲線圖譜。示出清晰的熔解曲線圖譜。 克隆性分析克隆性分析u 概念：確定一群體細(xì)胞是單克隆性的還是多概念：確定一群體細(xì)胞是單克隆性的還是多克隆性的克隆性的u 意義：正常組織和反應(yīng)性增生的病變是多克意義：正常組織和反應(yīng)性增生的病變是多

34、克隆起源，而絕大多數(shù)腫瘤是單克隆起源隆起源，而絕大多數(shù)腫瘤是單克隆起源診診斷與鑒別診斷意義斷與鑒別診斷意義克隆性分析的方法克隆性分析的方法u T T、B B細(xì)胞受體基因重排的克隆性分析細(xì)胞受體基因重排的克隆性分析u 基于基于X X染色體連鎖的體細(xì)胞克隆性分析染色體連鎖的體細(xì)胞克隆性分析u 細(xì)胞表型分析細(xì)胞表型分析u 體細(xì)胞突變位點(diǎn)分析體細(xì)胞突變位點(diǎn)分析u 染色體中病毒染色體中病毒DNADNA整合位點(diǎn)分析整合位點(diǎn)分析Rearrangement of the immunoglobulin heavy chain gene generates antigen recognition diversit

35、y in normal B-cells. Three areas of the germline IgH gene are moved and brought together during B-cell maturation, the V (variable), D (diversity) and J (joining) regions受體基因重排受體基因重排Site-Specific Recombination in Eukaryotes: Immunoglobulin Gene Rearrangement During Development2626個(gè)英文字母可以組成數(shù)以億萬計(jì)的單詞個(gè)英

36、文字母可以組成數(shù)以億萬計(jì)的單詞l A Al ATATl EATEATl BIRDBIRDl ABOUTABOUTl HISTORYHISTORYl WELCOMEWELCOMEl NOVEMBERNOVEMBERl PATHOLOGYPATHOLOGYl l IMMUNOHISTOCHEMISTRYIMMUNOHISTOCHEMISTRYl .l 多克隆性多克隆性l WELCOMEWELCOMEl WELCOMEWELCOMEl WELCOMEWELCOMEl WELCOMEWELCOMEl WELCOMEWELCOMEl WELCOMEWELCOMEl WELCOMEWELCOMEl WEL

37、COMEWELCOMEl WELCOMEWELCOMEl WELCOMEWELCOMEl WELCOMEWELCOMEl WELCOMEWELCOMEl 單克隆性單克隆性新的新的BIOMED-2BIOMED-2系統(tǒng)系統(tǒng)u歐洲淋巴瘤合作組織制定；歐洲淋巴瘤合作組織制定；u107107條引物；條引物；uT T、B B檢測：檢測：1414組項(xiàng)組項(xiàng)PCRPCR；u內(nèi)對照、腫瘤對照：共內(nèi)對照、腫瘤對照：共3030余組；余組；uInVivoScribe InVivoScribe 公司試劑；公司試劑；u3 3個(gè)工作日；個(gè)工作日；u篩選病例做，免疫組化能解決的不做。篩選病例做，免疫組化能解決的不做。BIOME

38、D-2BIOMED-2系統(tǒng)的優(yōu)勢系統(tǒng)的優(yōu)勢 部分解決部分解決假陰性假陰性的問題的問題u 假陰性的主要原因之一是設(shè)計(jì)的引物少，不能假陰性的主要原因之一是設(shè)計(jì)的引物少，不能涵蓋可能發(fā)生的各種重排。涵蓋可能發(fā)生的各種重排。u 如果把一個(gè)字母理解為一段引物的話，引物不如果把一個(gè)字母理解為一段引物的話，引物不夠，就相當(dāng)于字母的種類不夠，因?yàn)闆]有夠，就相當(dāng)于字母的種類不夠，因?yàn)闆]有2626個(gè)個(gè)字母就不能保證拼出各種單詞！字母就不能保證拼出各種單詞！u 方法似乎方法似乎“煩瑣煩瑣”了，但是確實(shí)科學(xué)合理了，了，但是確實(shí)科學(xué)合理了，而且實(shí)際應(yīng)用并沒有真正煩瑣多少，只是多做而且實(shí)際應(yīng)用并沒有真正煩瑣多少，只是多做

39、些些PCRPCR，多花些時(shí)間判斷！，多花些時(shí)間判斷！BIOMED-2BIOMED-2系統(tǒng)的局限系統(tǒng)的局限 本身并不能解決假陽性的問題本身并不能解決假陽性的問題u條帶必需在特定的預(yù)期的目標(biāo)位置上；條帶必需在特定的預(yù)期的目標(biāo)位置上；u條帶不是預(yù)期的假陽性條帶；條帶不是預(yù)期的假陽性條帶；u結(jié)果可重復(fù)；結(jié)果可重復(fù)；u對于多于兩個(gè)條帶時(shí)（如三個(gè)）是寡克隆、亞克隆對于多于兩個(gè)條帶時(shí)（如三個(gè)）是寡克隆、亞克隆的腫瘤性克隆演進(jìn)現(xiàn)象還是多克隆更要密切結(jié)合形的腫瘤性克隆演進(jìn)現(xiàn)象還是多克隆更要密切結(jié)合形態(tài)判斷；態(tài)判斷；u由于由于TCR-TCR-不同克隆之間基因重排片段的大小差異不同克隆之間基因重排片段的大小差異較小

40、，更易出現(xiàn)假陽性；較小，更易出現(xiàn)假陽性；u配合其它方法區(qū)別單克隆與多克?。好?xì)管凝膠電配合其它方法區(qū)別單克隆與多克隆：毛細(xì)管凝膠電泳、梯度凝膠電泳、單鏈構(gòu)象多態(tài)性、自動化熒光泳、梯度凝膠電泳、單鏈構(gòu)象多態(tài)性、自動化熒光分析技術(shù)等。分析技術(shù)等。BIOMED-2BIOMED-2系統(tǒng)沒有解決基因重排系統(tǒng)沒有解決基因重排克隆性分析的所有問題克隆性分析的所有問題u 重排基因中的突變和缺失問題重排基因中的突變和缺失問題u 基因重排不能替代免疫組化，進(jìn)一步的分型基因重排不能替代免疫組化，進(jìn)一步的分型還是依靠免疫組化還是依靠免疫組化u 仍然要強(qiáng)調(diào)四結(jié)合：臨床、形態(tài)、免疫表型、仍然要強(qiáng)調(diào)四結(jié)合：臨床、形態(tài)、免疫

41、表型、克隆性分析克隆性分析腫瘤分子病理診斷的應(yīng)用腫瘤分子病理診斷的應(yīng)用uu 腫瘤的分類腫瘤的分類uu 腫瘤診斷與早期診斷腫瘤診斷與早期診斷uu 腫瘤預(yù)后的判斷與檢測腫瘤預(yù)后的判斷與檢測uu 腫瘤個(gè)體化治療腫瘤個(gè)體化治療核酸分子診斷示例之一核酸分子診斷示例之一: FISH: FISH乳腺癌治療藥物乳腺癌治療藥物HerceptinHerceptin的選擇的選擇u HER-2HER-2陽性占乳腺癌的陽性占乳腺癌的25253030；對三苯氧；對三苯氧胺治療無效，甚至病情進(jìn)展（部分胺治療無效，甚至病情進(jìn)展（部分ERER、PRPR陽性不宜用三苯氧胺治療）；陽性不宜用三苯氧胺治療）；u 化療方案選擇：含蒽環(huán)

42、類抗生素敏感；環(huán)化療方案選擇：含蒽環(huán)類抗生素敏感；環(huán)甲氟聯(lián)合化療有抗性；單抗可逆轉(zhuǎn)腫甲氟聯(lián)合化療有抗性；單抗可逆轉(zhuǎn)腫瘤浸潤、轉(zhuǎn)移。瘤浸潤、轉(zhuǎn)移。u HER-2HER-2的檢測不是僅僅確定陽性或陰性，的檢測不是僅僅確定陽性或陰性，而是確定基因過表達(dá)（擴(kuò)增）的程度。而是確定基因過表達(dá)（擴(kuò)增）的程度。u 免疫組化：分級初篩免疫組化：分級初篩u FISHFISH確定：具有高度重復(fù)性和可靠性確定：具有高度重復(fù)性和可靠性Abnormal 2+Abnormal 3+Normal 0Normal Normal 1+Normal Abnormal lowamplificationAbnormal highamp

43、lification免疫組化與免疫組化與FISHFISH51比比2 20132 201352核酸分子診斷示例之二核酸分子診斷示例之二: FISH: FISH用于鑒別診用于鑒別診斷斷 Case 1 : BL A 21/M presented with neck node swelling, 5 ml aspirate taken from submandibular LN. Clinical: Reactive LN, infection? neoplastic?Diagnosis: Diagnosis: P Presence of t(8;14)(q24:q32)/MYC-IGH BLrese

44、nce of t(8;14)(q24:q32)/MYC-IGH BL FISH: MYC +, IGH +IGH-MYC +BCL2 BCL6 IGH break part +VeMYC break part +VeIGH-MYC fusion +Ve Histology: Relatively monomorphic intermediate sized lymphocytes with high N:C ratio, occasional large tingible body macrophages. IHC: CD20+, CD10+, BCL6+, MIB-1 90% CD3-, C

45、D30-, BCL2- Pre. Dx: High grade B-NHL. BL?核酸分子診斷示例之三核酸分子診斷示例之三 協(xié)助診斷協(xié)助診斷 詹某，女，詹某，女，5555歲，發(fā)熱，歲，發(fā)熱，皮疹，肝脾大，肝功能異常。皮疹，肝脾大，肝功能異常。頸部淋巴結(jié)切片，會診病例。頸部淋巴結(jié)切片，會診病例。IgIg與與TCRTCR重排克隆性分析重排克隆性分析1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30Case 斑某，男，斑某，男，4040歲，歲，發(fā)現(xiàn)頸部包塊發(fā)現(xiàn)頸部包塊1 1月，行頸部月，行

46、頸部淋巴結(jié)活檢。淋巴結(jié)活檢。CD3CD20IgIg與與TCRTCR重排克隆性分析重排克隆性分析1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 Case StudyCase Study1515歲，陰道出血，歲，陰道出血，HCGHCG沒有明顯沒有明顯增高。子宮內(nèi)容物送檢。增高。子宮內(nèi)容物送檢。核酸分子診斷示例之四核酸分子診斷示例之四: : 倍體分析與特定基因表達(dá)結(jié)合倍體分析與特定基因表達(dá)結(jié)合 由于由于B B超等檢測技術(shù)進(jìn)步，過去超等檢測技術(shù)進(jìn)步，過去1414周發(fā)周發(fā)現(xiàn)，現(xiàn)在提早到現(xiàn)，現(xiàn)

47、在提早到6 61212周，早期診斷帶來困周，早期診斷帶來困難。難。 過去認(rèn)為早期漏診也沒關(guān)系，現(xiàn)在過去認(rèn)為早期漏診也沒關(guān)系，現(xiàn)在臨床發(fā)現(xiàn)持續(xù)的滋養(yǎng)葉細(xì)胞疾病與固有的生臨床發(fā)現(xiàn)持續(xù)的滋養(yǎng)葉細(xì)胞疾病與固有的生物學(xué)特性相關(guān)，而與孕周關(guān)系不大，因此，物學(xué)特性相關(guān)，而與孕周關(guān)系不大，因此，診斷早期水泡狀胎塊是非常重要的。診斷早期水泡狀胎塊是非常重要的。Early complete mole: confusion with partial mole and hydropic villi. How to recognize? 早期完全性水泡狀胎塊與部分性水泡早期完全性水泡狀胎塊與部分性水泡狀胎塊及絨毛水腫的鑒別？狀胎塊及絨毛水腫的鑒別？ P57P57：來自父親的：來自父親的等位基因是沉默的，等位基因是沉默的，只有來自母親的等位只有來自母親的等位基因才表達(dá)。完全性基因才表達(dá)。完全性水泡狀胎塊水泡狀胎塊P57P57陰性。陰性。完全性與部分性的鑒別：完全