小儿急性白血病MTS1基因缺乏及点突变的研究

为探讨ＭＴＳ１基因突变与恶性血液病的关系，应用ＰＣＲ－ＳＳＣＰ和ＤＮＡ印迹方法检测３５例急性白血病（ＡＬ）患儿ＭＴＳ１基因改变。结果显示：急性淋巴细胞白血病（ＡＬＬ）缺乏（包括点突变）为２５．８％（８／１３）。Ｂ－ＡＬＬ纯合缺失为１６％（４／２５），Ｔ－ＡＬ为３３％（２／６）。点突变则两型各１例。结果证明：我国儿童ＡＬＬ ＭＴＳ１基因失活的存在，Ｔ－ＡＬ高于Ｂ－ＡＬＬ，点突变仅见于少数病例。该基     

应用PCR检测恶性淋巴瘤骨髓浸润及其临床意义

目的：检测恶性淋巴瘤（ＭＬ）的骨髓浸润（ＩＢＭ）并探讨其与临床分期、疗效和预后的关系。方法：以克隆性ＩｇＨ和ＴＣＲγ基因重排分别作为Ｂ和Ｔ细胞淋巴瘤克隆基因标志，应用ＰＣＲ基因扩增技术检测ＭＬ患者ＩＢＭ。结果：（１）３４份ＭＬ患者骨髓标本ＩＢＭ检出率为７０．６％（２４／３４），明显高于形态学检测方法（３８．６％，Ｐ＜０．０５）。（２）１９例形态学检测正常的非霍奇金淋巴瘤（ＮＨＬ）骨髓标本，ＩＢＭ阳性率５７．９％（１１／１９），其中８例Ｂ－ＮＨＬ（８／１４）检测到克隆性ＩｇＨ基因重排；５例同时还检测到克隆性ＴＣＲγ基因重排，２例Ｔ－ＮＨＬ（２／３）检测到克隆性ＴＣＲγ基因重排，无克隆性ＩｇＨ基因重排；２例免疫分型不明确ＮＨＬ患老中１例（１／２）检测到克隆性ＩｇＨ基因重排，无克隆性ＴＣＲ，基因重排。２例霍奇金病（ＨＤ）和１０例非淋巴系肿瘤骨髓ＩＢＭ均阴性。（３）Ⅲ，Ⅳ期ＮＨＬ患者ＩＢＭ检出率显著高于ＩＩ期（ｐ＜０．０１），初诊未治及复发患者也显著高于部分或完全缓解患者（Ｐ＜０．０１）。（４）ＩＢＭ阳性组ＮＨＬ２年死亡率５４．５％（６／１１），明显高于ＩＢＭ阴性（ｐ＜０．０５）。结论：ＰＣＲ方法检测ＩＢＭ有助于估     

黄曲霉毒素B_1单克隆抗体的制备和鉴定

以自己合成的黄曲霉毒素Ｂ＿１一人血清白蛋白（ＡＦＴＢ＿１－ＨＳＡ）偶联物免疫ＢＡＬＢ／ｃ鼠，应用杂交瘤技术，建立了两株能持续分泌抗ＡＦＴＢ＿１单克隆抗体（ＡＦＴＢ＿１ＭｃＡｂ）的杂交瘤细胞株，命名为ＩＢ５和２Ｆ１，接种ＢＡＬＢ／ｃ鼠腹腔均能诱发产生含特异性抗体的腹水。用间接竞争抑制ＥＬＩＳＡ鉴定ＭｃＡｂ的反应特异性，使ＭｃＡｂ与固相黄曲霉毒素Ｂ＿１一牛血清白蛋白（ＡＦＴＢ＿１－ＢＳＡ）结合产生５０％抑制所需的ＡＦＴＢ＿１、Ｂ＿２、Ｇ＿１、Ｇ＿２浓度依次为：１Ｂ５４．０、３６．９、２３．３、４０３．３ｎｇ／ｍｌ，２Ｆ１２．４、２．６、２．８、４。７ｎｇ／ｍｌ。     

成人急性淋巴细胞白血病分子生物学改变与临床治疗及预后关系的初步研究

应用多聚酶链反应技术检测了１４例急性淋巴细胞白血病（ＡＬＬ）患者的ｂｃｒ／ａｂｌ融合基因、ＴＣＲγ基因重排及ＩｇＨ基因重排，结果发现，ｂｃｒ／ａｂｌ融合基因阳性５例，阳性率３５．７％，其中４例为Ｂ－ＡＬＬ，１例ＴＣＲγ及ＩｇＨ基因重排均阴性。ｂｃｒ／ａｂｌ（＋）患者与ｂｃｒ／ａｂｌ（－）患者在临床表现方面无显著性差异，但ｂｃｒ／ａｂｌ（＋）患者治疗效果比ｂｃｒ／ａｂｌ（－）患者差。     

淋巴细胞性白血病／淋巴瘤bcl—2基因重排的研究

目的：探讨ｂｃｌ－２／ＩｇＨ基因重排的临床意义。方法：应用多聚酶链反应技术检测９种恶性淋巴瘤细胞系和不同类型淋巴系统恶性肿瘤临床标本中ｂｃｌ－２／ＩｇＨ基因重排情况。结果：２种细胞系,１例慢性淋巴细胞白血病及６例非霍奇金淋巴瘤（ＮＨＬ）有ｂｃｌ－２／ＩｇＨ基因重排,其中,３６．４％滤泡型ＮＨＬ及１８．２％弥漫性ＮＨＬ有重排。ｂｃｌ－２基因断裂点绝大多数（８／９）位于主要断裂区（ＭＢＲ）。结论：ｂｃ     

抗沙眼衣原体主要外膜蛋白种特异性单克隆抗体

用人工合成沙眼衣原体主要外膜蛋白（ＭＯＭＰ）氨基端１／４肽链免疫８周龄雄性ＢＡＬＢ／ｃ小鼠３只，第１４天用Ｌ１／４４０／Ｂｕ原体加强免疫，第２４天将免疫反应良好的１只小鼠脾细胞与ＮＳ－１瘤细胞融合。用１／４ＭＯＭＰ和Ｌ１原体抗原包被的聚丙乙烯板检测上清中抗体，淘汰与鹦鹉热衣原体种（ＥＡＥ株）、肺炎衣原体种（ＡＴＣＣＶＲ１３１０株）以及正常组织培养细胞（ＢＧＭＫ）有交叉反应的克隆，最后得到４株抗沙眼衣原体特异性单克隆抗体（ＭＡｂｓ），其抗体属ＩｓＧ１和ＩｇＧ２ａ亚类。微量免疫荧光试验（ｍｉｃｒｏ－ＩＦ）发现４株ＭＡｂｓ均与本实验室制备的沙眼衣原体Ｌ１、Ｌ２、Ａ、Ｂ、Ｃ、Ｅ原体及Ｌ２包涵体抗原发生反应，并与沙眼衣原体所有１５个标准血清型结合，其腹水ＭＡｂｓ滴度≥１：１２８００。免疫印迹试验显示４株ＭＡｂｓ均与沙眼衣原体分子量为４００００的ＭＯＭＰ反应。     

应用PT／PCR技术检测AML—M2的L1／ETO融合基因转录本

ｔ（８;２１）（ｑ２２;ｑ２２）易位是急性髓细胞白血病（ＡＭＬ）中最常见的染色体畸变之一,与ＡＭＬ－Ｍ２型密切相关,故ＭＩＣ分型会议将其命名为Ｍ２／ｔ（８;２１）。近年的研究发现ｔ（８;２１）易位分别累及２１号染色体上的ＡＭＬ１基因和８号染色体上的ＥＴ０基因,在ｄｅｒ（８）上形成一具有转录活性的ＡＭＬ１－ＥＴ０融合基因。本文建立了半筑巢式ＲＴ／ＰＣＲ技术,用以检测ＡＭＬ１／ＥＴ０融合基因转录本,并探讨该技术在Ｍ２／ｔ（８;２１）的诊断及微小残留病监测中的作用和意义。１　材料和方法１．１　研究对…     

造血干细胞移植治疗白血病和淋巴瘤

目的：探讨造血干细胞移植治疗白血病和淋巴瘤的疗效、移植过程中的并发症及其预防和移植后的植活证据及微小残余病灶检测等问题。方法：２例行同胞间异基因骨髓移植（Ａｌｌｏ－ＢＭＴ）、５例行自体外周血干细胞移植（ＡＰＢＳＣＴ）。采用ＣｓＡ＋ＭＴＸ预防ＧＶＨＤ;ＰＧＥ１＋潘生丁预防ＨＶＯＤ;以ＡＰＯＢ、ＳＭ７和Ｄ１Ｓ８０三个位点多态性分析作为植活证据;以ｂｃｒ／ａｂｌ、ＡＭＬ１－ＥＴＯ融合基因或ＩｇＨ基因重排     

影响糖尿病患者胆固醇水平的多因素分析(附80例临床分析)

本文观察８０例非胰岛素依赖型糖尿病（ＮＩＤＤＭ）患者的病程、糖化血红蛋白（ＧＨｂ）、甘油三脂（ＴＧ）、高密度脂蛋白－胆固醇（ＨＤＬ－Ｃ）、低密度脂蛋白－胆固醇（ＬＤＬ－Ｃ）、ＨＤＬ－Ｃ／ＬＤＬ－Ｃ、载脂蛋白Ａ１（ａｐｏＡ１）、载脂蛋白Ｂ１００（ａｐｏＢ１００）、ａｐｏＡ１／ａｐｏＢ１００、雌二醇（Ｅ２）、睾酮（Ｔ）、Ｅ２／Ｔ、尿白蛋白排泄率（ｕＡＥＲ）等因素对总胆固醇（ＴＣ）的影响，应用ＳＡＳ统计软件进行逐步回归分析。结果显示：ＴＣ与ＬＤＬ－Ｃ、ｕＡＥＲ显著正相关，与Ｅ２或Ｅ２／Ｔ显著负相关；而ｕＡＥＲ又与ＴＣ、ＧＨｂ、病程均显著正相关，提示ＮＩＤＤＭ患者胆固醇异常与脂蛋白代谢、性激素代谢、肾功能以及糖代谢等紊乱有关。     

单克隆抗体ELISA测定血浆富组氨酸糖蛋白的方法

建立用单克隆抗体（ＭｃＡｂ）酶联免疫吸附测定法（ＥＬＩＳＡ）测定血浆富组氨酸糖蛋白（ＨＲＧ）的方法。结果显示：ＭｃＡｂ３Ｃ６Ｆ３包被浓度为１０μｇ·ｍｌ－１，单抗酶标结合物（ＭｃＡｂ９Ｅ７Ｃ９ＨＲＰ）作１∶２００稀释时标准曲线最理想；最低检出限为１ｎｇ·ｍｌ－１，批内ＣＶ８２％，批间ＣＶ１１５％，回收率为８９１％，测定范围为２５～８０ｎｇ·ｍｌ－１；用该法测定４０例正常人血浆ＨＲＧ为（１１０３±９７）ｍｇ·Ｌ－１，２４例肝硬化患者血浆ＨＲＧ为（７９５±１２６）ｍｇ·Ｌ－１。该法的特异性强，且方便而准确。     

急性髓系白血病AML1/ETO融合基因检测及其临床意义

目的 了解急性髓系白血病中AML1／ETO融合基因表达情况及相关临床意义。方法 应用RT-PCR技术检测159例初发未治急性髓系白血病患者骨髓单个核细胞（MNC）AML1／ETO融合基因及R显带技术检测染色体,并对部分病人进行追踪检测及随访。结果 159例未经选择的初发急性髓系白血病病人中,31例（19．5％）有t（8;21）（q22;q22）易位,36例（22．6％）表达AML1／ETO融合基因,所有t（8;21）阳性的病人均存在AML1／ETO融合基因。AML1／ETO融合基因阳性病人缓解率为80．6％（29／36）,高于阴性病人（M,除外）的60．2％（56／93）;AML1／ETO阳性病人经治疗后转阴并持续阴性者预后好,而持续阳性者预后差,由阴性转阳性者可预示复发。14例持续缓解超过18个月的病人AML1／ETO融合基因仅1例为阳性。结论 （1）RT—PCR法检测AML1／ETO mRNA敏感、快速,定期检测可监测微小残留病变。（2）单次PCR法扩增AMLl／ETO融合基因优于巢式PCR法。     

GSTT1,GSTM1和NQO1基因多态性与急性髓系白血病发生及其重现染色体异常关系的研究

目的探讨GSTT1,GSTM1和NQO1基因多态性与原发性急性髓系白血病（AML）易感性及AML染色体核型异常的关系。方法228例AML和241名与患者无血缘关系的正常人群对照,用多重聚合酶链反应（PCR）方法检测GSTT1和GSTM1基因型,用PCR-限制性内切酶片段长度多态性（RFLP）方法分析NQO1基因型。结果AML患者GSTM1无效型（null）比例（62．3％）明显高于正常对照组（52．7％）,而GSTT1无效型（null）比例与正常对照组比较,差异无统计学意义。NQO1C609TC／T和T／T基因型比例AML患者（分别为53．1％和25．0％）、t（8;21）（q22;q22）／AML—ETO阳性患者（分别为64．3％和25．0％）、t（15;17）（q22;q11）／PML—RARα阳性患者（分别为57．1％和26．0％）明显高于正常对照组（分别为49．4％和13．7％）。NQO1C609TC／T和T／T基因型患t（8;21）（q22;q22）／AML-ETO阳性AML的相对危险性分别为4．487（95％CI：1．282—15．705）和6．293（95％CI：1．536—25．782）,NQO1C609TC／T和T／T基因型患t（15;17）（q22;q11）／PML—RARα阳性AML的相对危险性分别为2．531（95％CI：1．286—4．981）和4．149（95％CI：1．856—9．275）。结论NQO1C609TC／T和T／T基因型与我国成人原发性AML,特别是伴重现染色体异常t（8;21）（q22;q22）／AML—ETO阳性及t（15;17）（q22;q11）／PML—RARα阳性AML高度相关。     

Chromosomal changes detected by fluorescence in situ hybridization in patients with acute lymphoblastic leukemia

Objectives To investigate patients with acute lymphoblastic leukemia (ALL) for TEL/AML1 fusion,BCR/ABL fusion, MLL gene rearrangements, and numerical changes of chromosomes 4, 10, 17 and 21 by fluorescence in situ hybridization (FISH) and to determine the relationship and the significance of those findings.Methods Fifty-one American patients (34 men and 17 women) were included in this study. Of them there were 41 patients with pro-B cell type ALL, 9 with B cell type ALL and 1 with T cell type ALL.Chromosome metaphases of each sample were prepared according to standard protocols.Fluorescence in situ hybridization was performed using commercially available DNA probes, including whole chromosome painting probes, locus specific probes, specific chromosome centromere probes and dual color/multiple color translocation fusion probes. The digital image analysis was carried out using Cytovision and Quips FISH programs.Results An overall incidence of chromosomal anomalies, including t (9; 22 ), MLL gene rearrangements, t (12;21), and numerical chromosomal anomalies of chromosomes 4, 10, 17 and 21 was found in 33 patients (65%). Thirty-one of them were pediatric patients and two adults. The t(12;21) was the commonest chromosomal anomaly detected in this population; 14 out of the 45pediatric patients (31%) were positive for TEL/AML1 fusion, among which three had an additionalderivative 21[t (12;21) ], four had a deletion of 12p and two had an extra copy of chromosome 21.All 14 patients with positive TEL/AML1 fusion had ALL pre-B cell or B-cell lineage according to standard immunotyping. The percentage of cells with fusion signals ranged from 20% to 80%. All fourteen patients positive for TEL/AML1 gene fusion were mosaic. Three out of the 14 patients positive for the TEL/AML1 gene fusion were originally reported to be culture failures and none of the remaining eleven samples had been found to have chromosome 12 abnormalities by conventional cytogenetic techniques. All pediatric patients with pre-T or T cell lineage and the six adults were negative for TEL/AML1 fusion. One patient had double Philadelphia chromosomes, three had a rearranaement or a deletion of the MLL aene. one had t (4;11)and two had a deletion of the MLL One of the patients with an MLL deletion also had a large ring of chromosome 21, and r (21) was caused by AML1 gene tandemly duplicated at least five times. The second case with the MLL deletion was also unique, the patient had a t (12;21) as well. A total of 20 patients had numerical changes( gain or loss) of chromosomes 4, 10, 17 and 21. Eight patients were found to have trisomies of three or four different chromosomes. Interestingly, seven of these patients did not have TEL/AML1, BCR/ABL or the MLL aene rearranaement, one did have the TEL/AML1 aene fusion. Eleven patients with pro-B cell or B cell type ALL (9 children with ALL, 2 adults with ALL) had numerical changes of chromosome 21 (gain 1 or 2 chromosome 21 ), among them, 10 patients had no structural alteration of chromosome 21, and one was combined by t (12;21 ). Four patients had a monosomy of chromosome 17 and three out of these patients with monosomy 17 also had a fusion signal of TEL/AML1.     

MiR-130a is aberrantly overexpressed in adult acute myeloid leukemia with t(8;21) and its suppression induces AML cell death

Background: Emerging evidence has revealed that miRNAs can function as oncogenes or tumor suppressor genes in leukemia. The ectopic expression of miR-130a has been reported in chronic leukemia, but our understanding of the biological implications of miR-130a expression remains incomplete.

Methods: We quantified a cohort of de novo acute myeloid leukemia (AML) by bead-based miRNA and real-time quantitative PCR (Rq-PCR). The luciferase reporter gene assay was analyzed after the plasmid constructs which contain 5’-UTR of miR-130a and a Renilla luciferase reporter plasmid were transfected simultaneously into 293T cells. MTT and caspase 3/7 apoptosis assays were used to test cell viability and apoptosis.

Results: We identified miR-130a as significantly overexpressed in t(8;21) AML. Expression of miR-130a decreased significantly once patients with t(8;21) achieved complete remission, but increased sharply at the time of relapse. In patients with t(8;21) AML, KIT mutational status was associated with miR-130a expression—with higher expression associated with KIT activating mutations. Increased miR-130a expression in t(8;21) AML was associated with slightly worse event-free survival; however, no impact on overall survival was observed. Knockdown of AML1/ETO protein in the SKNO-1 cell line resulted in decrease of expression of miR-130a. Direct binding of AML1/ETO fusion protein with the promoter sequence of miR-130a was detected with luciferase reporter gene assay. Following miR-130a knockdown, SKNO-1 demonstrated increased sensitivity to etoposide.

Conclusions: Our data suggest that miR-130a is directly activated by AML1/ETO, and may act as a factor which is associated with leukemia burden, event-free survival, and chemotherapy sensitivity in t(8;21) AML.     

Complex t(8;13;21)(q22;q14;q22)--a novel variant of t(8;21) in a patient with acute myeloid leukemia (AML-M2)

Variants of the t(8;21)(q22;q22) involving chromosome 8, 21, and other chromosomes account for about 3% of all t(8;21)(q22;q22) in acute myeloid leukemia (AML) patients. We report a case of AML-M2 with t(8;13;21)(q22;q14;q22), not reported earlier. Using a dual-color fluorescence in situ hybridization (FISH) analysis with ETO and AML1 probes, we demonstrate an ETO/AML1 fusion signal on the derivative chromosome 8. Whole chromosome painting probes were used for chromosomes 8 and 13, to demonstrate the three-way translocation t(8;13;21)(q22;q14;q22). Involvement of chromosome region 13q14 has never been reported earlier, although region 13q12 as a variant in AML with t(8;21) has been reported earlier. The possible role of genes in this region in leukemogenesis, its response to the treatment and its clinical implications are discussed.     

成人NUP98融合基因阳性AML患者的临床特点及生物学特征

目的 探讨成人急性髓细胞白血病（AML）患者中核孔蛋白98（NUP98）融合基因阳性患者的临床特点及生物学特征,以及NUP98融合基因与AML常见融合基因、预后基因的共表达对AML预后的影响。 方法 收集四川大学华西医院2014年7月1日至2017年3月1日间住院的成人初发AML和骨髓增生异常综合征（MDS）患者的骨髓或外周血标本,检测NUP98融合基因,并检测AML患者染色体核型。将染色体11p15重排或NUP98相关融合基因阳性的AML患者作为研究组,此期间初诊的其余AML患者作为对照组,并将其分为低、中、高危对照组。通过对照研究,分析其血液学特点、完全缓解率（CR）、与预后基因的共表达率以及生存分析。 结果 样本总量为197例。共16例（8.1%）患者存在NUP98相关融合基因（即研究组）,发现我院首例NUP98-拓扑异构酶Ⅰ（TOP1）融合基因阳性AML。研究组患者按FAB分型,主要为M2和M5;研究组Fms样酪氨酸激酶-3（FLT3）-基因内部串联重复突变（ITD）发生率[31.25%（5/16）]和死亡率[80.00%（4/5）]高于对照组[发生率9.95%（19/181）,死亡率42.11%（8/19）,P<0.05];研究组诱导化疗获CR率为78.57%,高于总对照组及其中高、中危亚组（P<0.05）。研究组中位总生存期（OS）为13月,中位无白血病生存期（LFS）仅为5月。 结论 NUP98融合基因阳性AML易合并其它融合基因及预后基因的共表达,LFS和OS较短,尤其发生FLT3-ITD共表达时,死亡率高。     

常见白血病融合基因筛查在白血病诊断与分型中的意义

目的:分析病人白血病细胞染色体畸变涉及的86种融合基因和临床白血病类型的相关性,探讨常见融合基因筛查法在临床诊断和分型中的应用价值.方法:收集161例初发或者复发的白血病患者及8例骨髓增生异常综合征(MDS)患者的骨髓细胞,提取RNA,用32条特异性引物逆转录为cDNA,利用白血病29种染色体畸变形成的融合基因的86种mRNA剪接变异体引物,分8管进行多重RT-PCR,筛查白血病融合基因.结合临床状态和形态学观察了解融合基因与白血病类型的关系.结果:白血病中115例(71%)分别检测出10种白血病常见融合基因,包括AMLl/ETO、PML/RARα、PLZF/RARα、dupMLL、MLL/AF6、MLL/AF10、CBFβ/MYHll、BCR/ABL、Hoxll、Evil.其中52例慢性粒细胞白血病(CML)100%检出BCR/ABL;25例急性早幼粒细胞白血病(APL)中88%检出融合基因,其中21例APL检测出PML/RARα,1例APL检测出PLZF/RARα;AMLl/ETO阳性的17例急性白血病(AL)16例为FAB-M2亚型,1例为混合型白血病;CBFβ/MYH11阳性的4例AL 3例为FAB分型的M4,1例为M5,属于向粒单细胞系统分化的白血病.16例AL检测出MLL基因异常,其中MLL/AF6白血病均为FAB分型的M5,具有典型的原始单核细胞白血病的特征.17例急性淋巴细胞白血病(ALL)5例检测出BCR/ABL.8例MDS病人中2例检测出融合基因,其中AMLl/ETO阳性的MDS-RAEB很快发展为AML.结论:这种以多重RT-PCR为基础的白血病常见融合基因筛查法可以准确、快速而且可靠地确定白血病的分子类型,提供白血病诊断和治疗的依据.     

聚合酶链反应阵列同时定量检测37种白血病融合基因

目的建立一种同时定量检测多种白血病染色体易位形成的融合基因的荧光实时定量聚合酶链反应（real-time PCR）方法,了解阵列式PCR的应用可行性。方法组合使用82条引物,设立66个PCR平行管,同时定量检测37种白血病常见融合基因形成的125种剪切体和4种白血病常见的原癌基因活化。实时定量PCR采用Eva Green荧光染料法,用ABL基因做内参,用比较Ct法进行相对定量。用此方案对31例白血病患者标本进行检测,其中6例慢性粒细胞白血病（CML）患者做治疗前后或治疗过程中不同时间的对比检测,28人次与多重巢式PCR方案的检测结果进行对照。结果我们建立的PCR阵列方案有较大的线性检测范围（10^2～10^8拷贝／μl）,有较高的检测灵敏度（232拷贝／μl）和精确性;在31例患者标本的检测中,共检测到14种融合基因类型和所有4种原癌基因活化;检测到一例同时有5种融合基因阳性和两种原癌基因活化的患者;和多重巢式PCR检测结果的对比显示本方案灵敏度略低于巢式PCR,但差异没有统计学意义（P=0．009）;6例CML患者标本的检测显示治疗后患者标本中BCR／ABL融合基因及WT1和EVI1原癌基因表达量均呈现不同程度的降低,与临床治疗情况符合;基因定量分析的结果表明本方案能够对常见白血病融合基因和癌基因活化情况进行定量分析,在白血病的诊断及疗效监测中有应用价值。结论PCR阵列法同时定量检测多种白血病融合基因适于白血病初诊患者融合基因的筛查及微小残留病（MRD）的检测,对于检测同时有多种融合基因存在的病例及治疗过程中融合基因的变异情况更为有用。     

实时定量聚合酶链式反应与巢式聚合酶链式反应在 289例白血病融合基因检测中的比较

 目的 比较实时定量聚合酶链式反应（RT-PCR）与巢式聚合酶链式反应（nPCR）在白血病融合基因BCR/ABL、PML/RARa及AML1/ETO检测中结果。方法 分别应用RT-PCR与nPCR对289例急慢性白血病初诊和完全缓解的患者BCR/ABL、PML/RARa及AML1/ETO融合基因进行检测,结合患者骨髓细胞形态学及临床转归予以分析。结果 46例初诊慢性粒细胞白血病（CML）患者中,RT-PCR法与nPCR法检测BCR/ABL融合基因阳性率分别为95.7%和93.5%（P=0.997）;69例完全缓解CML患者中,RT-PCR法与nPCR法检测阳性率分别为78.4%和58.1%（P<0.005）。32例初诊急性早幼粒细胞白血病（APL）患者中,RT-PCR法与nPCR法检测PML/RARa融合基因阳性率分别为93.8%和90.6%（P=0.996）;114例完全缓解APL患者中,RT-PCR法与nPCR法检测阳性率分别为12.3%和7.0%（P<0.025）。12例初诊急性粒细胞白血病M2（ANLL-M2）患者中,RT-PCR法与nPCR法检测AML1/ETO融合基因阳性率分别为50.0%和50.0%（P=1.0）;16例完全缓解ANLL-M2患者中,RT-PCR法与nPCR法检测阳性率分别为50.0%和12.5%（P<0.05）。结论 RT-PCR较nPCR更为敏感而准确,应用RT-PCR可以提高微小残留病的检出率,为临床诊断和治疗提供更为有效的分子生物学依据。     

双色双融合荧光原位杂交检测成人急性白血病PML/RARα、MLL、AML1/ETO基因重排

目的探讨双色双融合荧光原位杂交技术（DC-DF-FISH）在成人急性白血病（Acute Leukemia,AL）中的应用价值。方法应用DC-DF-FISH检测我院26例AL患者相应的AML1/ETO、MLL、PML/RARa融合基因,并与常规R-显带技术进行比较。结果 26例AL患者中,R-显带发现4例存在标记染色体,检出率15.4%（4/26）,17例为正常核型和其他异常核型,未检出包含11q23染色体异常,DC-DF-FISH检测发现8例特异目的基因为阳性,包括R-显带检测出的4例,检出阳性率30.8%（8/26）。结论双色双融合荧光原位杂交技术是检测成人急性白血病PML/RARα、MLL、AML1/ETO基因重排的可靠方法,适用于AL的诊断、疗效判定及微小残留病灶的检测。