目的探讨多聚酶链反应(PCR)检测免疫球蛋白重链基因重排在原发性胃肠道淋巴组织增生性疾病中的良恶性鉴别中的应用价值。方法选取存档蜡块45例,采用HE常规染色及SP免疫组化方法进行组织学分类,运用PCR半巢式扩增免疫球蛋白重链基因重排...目的探讨多聚酶链反应(PCR)检测免疫球蛋白重链基因重排在原发性胃肠道淋巴组织增生性疾病中的良恶性鉴别中的应用价值。方法选取存档蜡块45例,采用HE常规染色及SP免疫组化方法进行组织学分类,运用PCR半巢式扩增免疫球蛋白重链基因重排,2%琼脂糖凝胶电泳检测PCR产物。结果Ig H FR3A和Ig H FR3A+FR2A在MALT淋巴瘤的阳性检测率分别为60.7%(17/28)和75.0%(21/28),在DLBCL中的阳性检测率分别为60.0%(9/15)和73.3%(11/15),在MCL中的阳性检测率分别为100.0%(1/1)和100.0%(1/1)。Ig H FR3A在胃肠道淋巴瘤中的阳性检测率为60.0%。Ig H FR3A联合Ig H FR2Ad在胃肠道淋巴瘤中的阳性检出率为73.3%(33/45)。结论采用PCR方法检测免疫球蛋白重链基因重排能够作为淋巴瘤诊断的有效辅助手段,有助于对原发性胃肠道淋巴组织增生性疾病的良恶性做出正确的诊断。展开更多
We have recently developed a novel Immunoglobulin heavy chain gene rearrangement (IgH-R) assay that combines polymerase chain reaction (PCR) amplification and analysis in the same closed capillary tube using the Light...We have recently developed a novel Immunoglobulin heavy chain gene rearrangement (IgH-R) assay that combines polymerase chain reaction (PCR) amplification and analysis in the same closed capillary tube using the LightCycler System. IgHR can be identified by DNA melting curve analysis within 40 minutes after DNA preparation and amplification. To test the clinical utility of this new IgH-R assay for rapidly diagnosing cutaneous B-cell lymphomas, we prospectively analyzed 44 formalin-fixed, paraffin-embedded tissues suspected of Bcell malignant lymphoma: skin (n = 31), lymph node (n = 7), stomach (n = 3), spleen (n = 1), colon (n = 1), and soft tissue (n = 1). We detected IgH-R in 12 DNA samples, including 8 skin biopsies, with the following diagnoses: B-cell chronic lymphocytic leukemia (n = 4), extranodal marginal zone Bcell lymphoma (n = 4), diffuse large B-cell lymphoma (n = 2), Burkitt lymphoma (n = 1), and precursor B-lymphoblastic lymphoma(n = 1). DNAmelting curve analysis, comparedwith polyacrylamide gel electrophoresis, achieved a sensitivity equal to 92.3%and a specificity equal to 100%. There was a single false negative result because DNA melting curve analysis could not detect less than 10.0%clonal B-cells. We conclude that this new, rapid PCR assay for detecting IgH-R based on DNA melting curve analysis can be clinically useful for confirming the initial diagnosis of B-cell malignant lymphoma.展开更多
文摘目的探讨多聚酶链反应(PCR)检测免疫球蛋白重链基因重排在原发性胃肠道淋巴组织增生性疾病中的良恶性鉴别中的应用价值。方法选取存档蜡块45例,采用HE常规染色及SP免疫组化方法进行组织学分类,运用PCR半巢式扩增免疫球蛋白重链基因重排,2%琼脂糖凝胶电泳检测PCR产物。结果Ig H FR3A和Ig H FR3A+FR2A在MALT淋巴瘤的阳性检测率分别为60.7%(17/28)和75.0%(21/28),在DLBCL中的阳性检测率分别为60.0%(9/15)和73.3%(11/15),在MCL中的阳性检测率分别为100.0%(1/1)和100.0%(1/1)。Ig H FR3A在胃肠道淋巴瘤中的阳性检测率为60.0%。Ig H FR3A联合Ig H FR2Ad在胃肠道淋巴瘤中的阳性检出率为73.3%(33/45)。结论采用PCR方法检测免疫球蛋白重链基因重排能够作为淋巴瘤诊断的有效辅助手段,有助于对原发性胃肠道淋巴组织增生性疾病的良恶性做出正确的诊断。
文摘We have recently developed a novel Immunoglobulin heavy chain gene rearrangement (IgH-R) assay that combines polymerase chain reaction (PCR) amplification and analysis in the same closed capillary tube using the LightCycler System. IgHR can be identified by DNA melting curve analysis within 40 minutes after DNA preparation and amplification. To test the clinical utility of this new IgH-R assay for rapidly diagnosing cutaneous B-cell lymphomas, we prospectively analyzed 44 formalin-fixed, paraffin-embedded tissues suspected of Bcell malignant lymphoma: skin (n = 31), lymph node (n = 7), stomach (n = 3), spleen (n = 1), colon (n = 1), and soft tissue (n = 1). We detected IgH-R in 12 DNA samples, including 8 skin biopsies, with the following diagnoses: B-cell chronic lymphocytic leukemia (n = 4), extranodal marginal zone Bcell lymphoma (n = 4), diffuse large B-cell lymphoma (n = 2), Burkitt lymphoma (n = 1), and precursor B-lymphoblastic lymphoma(n = 1). DNAmelting curve analysis, comparedwith polyacrylamide gel electrophoresis, achieved a sensitivity equal to 92.3%and a specificity equal to 100%. There was a single false negative result because DNA melting curve analysis could not detect less than 10.0%clonal B-cells. We conclude that this new, rapid PCR assay for detecting IgH-R based on DNA melting curve analysis can be clinically useful for confirming the initial diagnosis of B-cell malignant lymphoma.
