Nowadays the role of genetic findings in determining the diagnosis,therapy and prognosis of acute myeloid leukemia(AML) has become more valuable.To improve and validate the detection of clonal chromosomal aberrations ...Nowadays the role of genetic findings in determining the diagnosis,therapy and prognosis of acute myeloid leukemia(AML) has become more valuable.To improve and validate the detection of clonal chromosomal aberrations in leukemia,we designed a combined application of karyotyping with multiplex reverse transcription-polymerase chain reaction(RT-PCR) and fluorescence in situ hybridization(FISH),and addressed the expression and distribution of fusion genes among the subtypes of Chinese adult patients with de novo AML.Multiplex RT-PCR assays were performed on 477 samples from newly diagnosed AML patients,and cytogenetic data were obtained from 373 of them by R or G banding techniques and those in some cases were confirmed by FISH.The PCR products in some suspected cases were tested by two-directional sequencing.The results showed that except unqualified samples,fusion genes were detected by multiplex RT-PCR in 211 of 474 patients(44.51%),including AML1-ETO,CBFβ-MYH11,PML-RARα,PLZF-RARα,NPM-RARα,MLL rearrangements,BCR-ABL,DEK-CAN,SET-CAN,TEL-PDGFR,TLS-ERG,AML1-MDS1(EVI-1).In 373 patients,who took both multiplex RT-PCR and karyotype analysis,the detection rate of chromosomal aberrations by using multiplex RT-PCR and karyotyping was 160/373(42.89%) and 179/373(47.98%) respectively,and the combination could optimize the detection rate of clonal genetic abnormalities to 216/373(57.90%).The PCR results from 11 cases 'normal' in karyotyping but abnormal in RT-PCR for MLL rearrangements were confirmed by two-directional sequencing.It is concluded that karyotype studies remain the cornerstone for genetic testing;conventional cytogenetics and molecular-based methods are complementary tests for the detection of clonal genetic aberrations in AML,especially for the cryptic or submicroscopic aberrations.Once a genetic marker has been identified by combined analysis,it could be used to monitor residual disease during/after chemotherapy,by quantitative RT-PCR and/or FISH.展开更多
Objective: Using nested reverse transcription-polymerase chain reaction(Nested RT-PCR) to test the mRNA level in peripheral blood CK19 and MUC-1 in colorectal cancer patients and it's clinical significance, to dis...Objective: Using nested reverse transcription-polymerase chain reaction(Nested RT-PCR) to test the mRNA level in peripheral blood CK19 and MUC-1 in colorectal cancer patients and it's clinical significance, to discuss the feasibility of colorectal carcinoma micro-metastasis detection of molecular markers. Methods: The expression level was detected by nested RT-PCR in 20 healthy people, 20 patients with colorectal adenoma and 90 cases of patients with colorectal cancer disease peripheral blood CK19 mRNA and MUC-1 mRNA. Results: The positive expression rate of CK19 mRNA and MUC-1 mRNA were: 58.89%(53/90) and 52.22%(47/90). No CK19 mRNA healthy people 20 cases in the control group in the peripheral blood, the expression of MUC-1 mRNA in 12 cases, the expression rate of 60%(12/20). In 20 cases of colorectal adenoma diseases have the expression of CK19 mRNA in 1 cases, the expression rate of 5%(1/20), the expression of MUC-1 mRNA in 10 cases, the expression rate of 50%. Patients with colorectal cancer CK19 mRNA, MUC-1 mRNA expression rate was significantly correlated with tumor staging, the degree of differentiation of the tumor cells and tumor metastasis(P < 0.05). Conclusion: Marker CK19 mRNA as the detection of micro-metastasis in peripheral blood of patients with colorectal cancer has good sensitivity and specificity, but CK19 mRNA, MUC-1 mRNA can be used to judge the prognosis of patients with colorectal cancer index.展开更多
文摘Nowadays the role of genetic findings in determining the diagnosis,therapy and prognosis of acute myeloid leukemia(AML) has become more valuable.To improve and validate the detection of clonal chromosomal aberrations in leukemia,we designed a combined application of karyotyping with multiplex reverse transcription-polymerase chain reaction(RT-PCR) and fluorescence in situ hybridization(FISH),and addressed the expression and distribution of fusion genes among the subtypes of Chinese adult patients with de novo AML.Multiplex RT-PCR assays were performed on 477 samples from newly diagnosed AML patients,and cytogenetic data were obtained from 373 of them by R or G banding techniques and those in some cases were confirmed by FISH.The PCR products in some suspected cases were tested by two-directional sequencing.The results showed that except unqualified samples,fusion genes were detected by multiplex RT-PCR in 211 of 474 patients(44.51%),including AML1-ETO,CBFβ-MYH11,PML-RARα,PLZF-RARα,NPM-RARα,MLL rearrangements,BCR-ABL,DEK-CAN,SET-CAN,TEL-PDGFR,TLS-ERG,AML1-MDS1(EVI-1).In 373 patients,who took both multiplex RT-PCR and karyotype analysis,the detection rate of chromosomal aberrations by using multiplex RT-PCR and karyotyping was 160/373(42.89%) and 179/373(47.98%) respectively,and the combination could optimize the detection rate of clonal genetic abnormalities to 216/373(57.90%).The PCR results from 11 cases 'normal' in karyotyping but abnormal in RT-PCR for MLL rearrangements were confirmed by two-directional sequencing.It is concluded that karyotype studies remain the cornerstone for genetic testing;conventional cytogenetics and molecular-based methods are complementary tests for the detection of clonal genetic aberrations in AML,especially for the cryptic or submicroscopic aberrations.Once a genetic marker has been identified by combined analysis,it could be used to monitor residual disease during/after chemotherapy,by quantitative RT-PCR and/or FISH.
文摘Objective: Using nested reverse transcription-polymerase chain reaction(Nested RT-PCR) to test the mRNA level in peripheral blood CK19 and MUC-1 in colorectal cancer patients and it's clinical significance, to discuss the feasibility of colorectal carcinoma micro-metastasis detection of molecular markers. Methods: The expression level was detected by nested RT-PCR in 20 healthy people, 20 patients with colorectal adenoma and 90 cases of patients with colorectal cancer disease peripheral blood CK19 mRNA and MUC-1 mRNA. Results: The positive expression rate of CK19 mRNA and MUC-1 mRNA were: 58.89%(53/90) and 52.22%(47/90). No CK19 mRNA healthy people 20 cases in the control group in the peripheral blood, the expression of MUC-1 mRNA in 12 cases, the expression rate of 60%(12/20). In 20 cases of colorectal adenoma diseases have the expression of CK19 mRNA in 1 cases, the expression rate of 5%(1/20), the expression of MUC-1 mRNA in 10 cases, the expression rate of 50%. Patients with colorectal cancer CK19 mRNA, MUC-1 mRNA expression rate was significantly correlated with tumor staging, the degree of differentiation of the tumor cells and tumor metastasis(P < 0.05). Conclusion: Marker CK19 mRNA as the detection of micro-metastasis in peripheral blood of patients with colorectal cancer has good sensitivity and specificity, but CK19 mRNA, MUC-1 mRNA can be used to judge the prognosis of patients with colorectal cancer index.
