Background: A dominant T- cell clone can be detected by polymerase chain reaction (PCR) in 40- 90% of cutaneous samples from patients with cutaneous T- cell lymphoma (CTCL). Materials and methods: From 1996 to 2003 we...Background: A dominant T- cell clone can be detected by polymerase chain reaction (PCR) in 40- 90% of cutaneous samples from patients with cutaneous T- cell lymphoma (CTCL). Materials and methods: From 1996 to 2003 we analysed 547 cutaneous biopsies performed to exclude CTCL (mycosis fungoides, MF/Sé zary syndrome, SS). The final diagnosis was benign inflammatory disease (BID) in 353 samples (64.5% )- and CTCL in 194 (35.5% ). T- cell receptor (TCR)- γ gene rearrangement was studied by using a multiplex PCR/heteroduplex (HD) analysis. The PCR results were correlated with the clinical picture, the histological pattern and the presence of T- cell lineage antigen loss, using univariate and multivariate logistic regression analyses. Objective: To determine the sensitivity and specificity of the multiplex PCR/HD analysis and to identify which are the clinical, histopathological or immunophenotypical features significantly associated with a positive T- cell clonality. Results: A clonality was demonstrated in 83.5% of CTCL and in 2.3% of BID (P < 0.001). A significantly higher percentage of clonal cases was associated with the cutaneous T- score (71.4% in T1, 76.1% in T2 and 100% in nodular and erythrodermic MF samples) and with the presence of a T- cell lineage antigen loss (93.9% vs. 77.4% ). Moreover, clonality was closely related to an increase in the histopathological score (51.3% in the samples with a score < 5, compared with 92% in the lesions with ≥ 5). No significant difference in the percentage of clonal cases was found between T1/T2 and T3/T4 lesions with a histopathological score ≥ 5. The multivariate logistic regression showed that the density and extent of the cell infiltrate, the degree of epidermotropism and the presence of cytological atypia share an independent predictive value for clonality in T1/T2 samples, even if the highest odds ratios (3.6) were associated with the density of the cell infiltrate. The disease course of T1/T2 patients was analysed according to the PCR findings. All the PCR- negative patients showed a long- standing stable disease course; on the other hand, a disease progression occurred in 12/87 (13.8% ) positive patients. Conclusions: The multiplex PCR/HD analysis is associated with a high diagnostic accuracy (92.7% ) in CTCL patients. The finding of a clonal T- cell rearrangement is more closely associated with the histological pattern (in particular with the density and extent of the cell infiltrate) rather than with the MF cutaneous T- score or immunophenotype.展开更多
【目的】比较单链构象多态性分析(SSCP)和异源双链分析(HA)在基因突变筛查中的意义。【方法】在帕金森病人和正常对照中,同时应用这两种实验方法筛查α-共核蛋白基因3、4外显子突变,电泳结果由测序考证。【结果】对照组第29号标本3号外...【目的】比较单链构象多态性分析(SSCP)和异源双链分析(HA)在基因突变筛查中的意义。【方法】在帕金森病人和正常对照中,同时应用这两种实验方法筛查α-共核蛋白基因3、4外显子突变,电泳结果由测序考证。【结果】对照组第29号标本3号外显子 SSCP 存在单链泳动异常。但 HA 检测无双链泳动异常,测序证实无基因突变。其它标本 SSCP 和 HA 检测均未发现异常泳动。【结论】将单链构象分析和异源双链分析相结合应用于基因突变筛查,既不用增加试剂、设备和技术难度,又可以优势互补,利于提高检测的准确性。展开更多
选用桑萎缩病(M u lberry dw arf,MD)、枣疯病(Ju jube w itches-broom,JW B)、酸枣丛枝病(W ild ju jube w itches-broom,W JW B)、泡桐丛枝病(Pau lownia w itches-broom,PaW B)和苦楝丛枝病(Chinaberry tree w itches-broom,CW B)5...选用桑萎缩病(M u lberry dw arf,MD)、枣疯病(Ju jube w itches-broom,JW B)、酸枣丛枝病(W ild ju jube w itches-broom,W JW B)、泡桐丛枝病(Pau lownia w itches-broom,PaW B)和苦楝丛枝病(Chinaberry tree w itches-broom,CW B)5种不同植物植原体和来源于3个不同地区PaW B和JW B材料进行16S rDNA和23S rDNA PCR扩增、异源双链迁移率分析(HMA)、PCR产物的RFLP分析和16S rDNA的克隆和测序等比较研究,建立了一种快速确定未知植原体种类和分类地位的分子鉴别与鉴定优化程序;并可对田间采集的各种植物植原体样品进行快速鉴定和鉴别。16S rDNA PCR产物HMA分析结果显示,JW B与CW B、MD和PaW B皆可形成明显的异源杂交双链;而CW B、MD和PaW B植原体之间未能形成异源双链。JW B和PaW B不同地区样品之间、JW B和W JW B之间也未发现异源杂交双链的形成。而23S rDNA PCR产物HMA分析则可以将MD与PaW B区分开。进一步对未知分类地位的CW B序列测定及与其它植原体16S rDNA的RFLP和同源性比较结果显示,CW B与PaW B同源性为99.5%,其中与MD的同源性高达99.7%,因而应将CW B归为翠菊黄花组16S r I-B,16S r I-B(rp-B)。展开更多
文摘Background: A dominant T- cell clone can be detected by polymerase chain reaction (PCR) in 40- 90% of cutaneous samples from patients with cutaneous T- cell lymphoma (CTCL). Materials and methods: From 1996 to 2003 we analysed 547 cutaneous biopsies performed to exclude CTCL (mycosis fungoides, MF/Sé zary syndrome, SS). The final diagnosis was benign inflammatory disease (BID) in 353 samples (64.5% )- and CTCL in 194 (35.5% ). T- cell receptor (TCR)- γ gene rearrangement was studied by using a multiplex PCR/heteroduplex (HD) analysis. The PCR results were correlated with the clinical picture, the histological pattern and the presence of T- cell lineage antigen loss, using univariate and multivariate logistic regression analyses. Objective: To determine the sensitivity and specificity of the multiplex PCR/HD analysis and to identify which are the clinical, histopathological or immunophenotypical features significantly associated with a positive T- cell clonality. Results: A clonality was demonstrated in 83.5% of CTCL and in 2.3% of BID (P < 0.001). A significantly higher percentage of clonal cases was associated with the cutaneous T- score (71.4% in T1, 76.1% in T2 and 100% in nodular and erythrodermic MF samples) and with the presence of a T- cell lineage antigen loss (93.9% vs. 77.4% ). Moreover, clonality was closely related to an increase in the histopathological score (51.3% in the samples with a score < 5, compared with 92% in the lesions with ≥ 5). No significant difference in the percentage of clonal cases was found between T1/T2 and T3/T4 lesions with a histopathological score ≥ 5. The multivariate logistic regression showed that the density and extent of the cell infiltrate, the degree of epidermotropism and the presence of cytological atypia share an independent predictive value for clonality in T1/T2 samples, even if the highest odds ratios (3.6) were associated with the density of the cell infiltrate. The disease course of T1/T2 patients was analysed according to the PCR findings. All the PCR- negative patients showed a long- standing stable disease course; on the other hand, a disease progression occurred in 12/87 (13.8% ) positive patients. Conclusions: The multiplex PCR/HD analysis is associated with a high diagnostic accuracy (92.7% ) in CTCL patients. The finding of a clonal T- cell rearrangement is more closely associated with the histological pattern (in particular with the density and extent of the cell infiltrate) rather than with the MF cutaneous T- score or immunophenotype.
文摘【目的】比较单链构象多态性分析(SSCP)和异源双链分析(HA)在基因突变筛查中的意义。【方法】在帕金森病人和正常对照中,同时应用这两种实验方法筛查α-共核蛋白基因3、4外显子突变,电泳结果由测序考证。【结果】对照组第29号标本3号外显子 SSCP 存在单链泳动异常。但 HA 检测无双链泳动异常,测序证实无基因突变。其它标本 SSCP 和 HA 检测均未发现异常泳动。【结论】将单链构象分析和异源双链分析相结合应用于基因突变筛查,既不用增加试剂、设备和技术难度,又可以优势互补,利于提高检测的准确性。
