Research on AML1/ETO fusion gene on 76 patients diagnosed with acute myelogenous leukemia

Background: Chromosome mutation type t(8;21) has quite a high frequency in acute myelogenous leukemia, which accounted for about 15% among adult patients. From 2001, the WHO has a new classification for acute myelogenous leukemia based on genetic mutations. Form had AML1/ETO were arranged into genetic mutation group with better prognosis and ability to fully recover after chemotherapy with a high dose of cytarabin. Objective: Study AML1/ETO fusion gene on the patients diagnosed with Acute Myelogenous Leukemia (AML), as well as the clinical features and some haematologic parameters of the AML1/ETO positive group. Subject and methods: 76 patients with AML were treating in the National Institute of Hematology & Blood Transfusion and the Department of Hematology & Blood Transfusion of Bach Mai Hospital from April 2007 to July 2008. These patients were studied for clinical examination, morphology and RNA were extracted from leukemic cells and PCR for AML1/ETO fusion transcript was performed. Results and conclusions: The incidence of AML1/ETO positive in the AML patients was 24%. The incidence of AML1/ETO positive in AML-M2 was 28%. In the AML1/ETO positive group: median age was 26.94+/-9.22; rate of severe anemia, hemorrhage, fever, infection, hepatomegaly, splenomegaly, lymphadenopathy and gum hypertrophy was 44%, 33%, 28%, 11%, 44%, 28%, 17% and 6%, respectively. Median hemoglobin, WBC, platelet, bone marrow cell count, % blast in peripheral blood and in bone marrow was 84.41+/-28.97 g/l, 29.42+/-31.36 g/l, 42.12+/-33.83 g/l, 215.93+/-134.42 g/l, 56.21+/-26.58% and 65.14+/-16.12%, respectively.

Study PML/RAR alpha fusion gene on 21 patients diagnosed with acute promyelocytic leukemia (AML \u2013 M3)

Background: In Vietnam, there are a number of studies on the application of ATRA in treating acute promyelocytic leukemia (AML \u2013 M3) but they have still faced with certain difficulties. Objectives: (1). Study PML/RAR alpha fusion gene on the patients diagnosed with AML \u2013 M3. (2). Study the index of hematology of the PML/RAR alpha positive group. Subject and Method: 21 patients with acute promyelocytic leukemia (M3) were studied. All patients were examined with morphology, coagulation and cytogenetic tests and RNA were extracted from leukemic cells and PCR for PML/RAR alpha fusion transcript. Result and conclusion: PML/RAR alpha positive in 67% including 4 patients which were not discovered t(15; 17) by cytogenetic technique. Rates of three subtype (bcr1, bcr2 and bcr3) of PML/RAR alpha were 7 patients (50%), 3 patients (21,5%) and 4 patients (28,5%), respectively. WBC and bone marrow cells of PML/RAR alpha positive group were 5.08+/-3.87 and 155.82+/-106.21. D \u2013 Dimer level was 1954.89+/-1575.28; 93% of patients in the PML/RAR alpha positive group had DIC.

Application PCR technique for analysis of fusion gene transcripts in the acute myelogenous leukemia

Background: In recent years, Vietnam has applied four methods (morphology, cell chemistry, immune marker classification, cyto genetic) in diagnosis and used multi-chemotherapy in treatment for acute myelogenous leukemia (AML)\r\n', u'Objectives: To initially determine some fusion gene transcripts in the acute myelogenous leukemia patients by applying PCR technique. Subject and method: The study included 19 patients with acute myelogenous leukemia treated in National Institute of Hematology and Blood Transfusion and Bachmai Hospital from April 2007 to August 2007. RNA were extracted from leukemic cells and PCR for AML1/ETO, CBFP/MYH11, PMR/RARa fusion transcript was done. Results: Number of male patients was 6 (32%), female patients was 13 (68%). The average age of these patients was 32.67 \xb113.62. There were three M4, M4eo patients with AML1/ETO gene (accounting for 16%), two M2, M4 patients with CBF/MYH1 gene and type F of genetic modification accounting for 11%), two M3 patients with PMR/RAR\u03b1 and Bcr3 of genetic modification (accounting for 11%). Conclusion: Results of the study did not differ significantly from other researches in the world. This study showed the need of applying the PCR technique in determining fusion gene transcript together with traditional cyto-genetic method.\r\n', u'\r\n', u'

Application of Elisa technique for detection anti \ufffd?HLA antibodies after solid organ transplantation

Background: Anti \ufffd?HLA (Human Leukocyte Antigen) antibody is result of immunization in allotransplantation. Organ transplantation is one of the great scientific achievements of the medicine. However, it is difficult to have the perfect harmony of HLA group. Inevitable consequence is the graft will be eliminated by the immune process. In Vietnam, organ transplantation was a relatively new specialty and there was not much research on evaluation immune process after transplantation. Objectives: To determine the rate of present of anti \ufffd?HLA antibody on transplant patients, and the role of post \ufffd?transplant anti \ufffd?HLA antibodies on long \ufffd?term graft function. Subjects and method: ELISA technique was used to analysis 31 blood samples of 31 patients who were transplanted organs at 103 military hospital and Cho Ray hospital from May 2000 to July 2007. This was a retrospective and described cross-sectional study on theclinical records. Results:The rate of anti \ufffd?HLA antibodies was 35.5%. The present of anti \ufffd?HLA antibodies of transplant patients had negative impact on graft function. Conclusion: The detection of anti \ufffd?HLA antibodies by ELISA in the post transplant period may be a high confident and sensitive technique for follows up graft function.

Studying antibodies against antigens of neutrophils in the multitransfused patients

Background: Patients who received multiple transfusions of blood and blood products may produce antibodies against antigens of erythrocytes, leukocytes, platelets etc, resulting in many clinical implications. Objectives: To detect frequencies of antineutrophil antibodies in multitransfused patients at National Institute of Hematology and Blood Transfusion (NIHBT). Subjects and methods: The study was conducted on 30 multitransfused patients. Among them there were 12 with thrombocytopenia and 18 with aplastic anemia. Results: 6 cases had anti - neutrophil antibodies, of which 5 had more than 5 times of transfusion, 4 with aplastic anemia and 2 with thrombocytopenia. The sera were further tested with neutrophil panel, revealing 4 samples with anti - NA 1 (13.3%) and 1 sample with anti - NA2 (3.3%). The frequency of anti - neutrophil antibodies in multitransfused patients at IHBT in the study is 20%. Conclusion: Frequency of anti-NA1 was higher than anti-NA2 in multitransfused patients at NIHBT and directly proportional by frequency of NA1 and NA2 antigens in this group. The technical process to identify and classify antineutrophil antibodies in this study can be applied for patients who received multiple transfusions of blood and blood products in Viet Nam

Application of FISH technique for detection of fusion gene ABL/BCR in chronic myelogenous leukemia

Background: Detection of BCR/ABL fusion gene has important significance in diagnosing and monitoring response to therapy in chronic myeloid leukemia. Objective: Application of FISH (Fluorescence In Situ Hybrydization) technique for detection of abl/bcr fusion gene in chronic myelogenous leukemia. Subjects and methods: The study included 10 patients of chronic myelogenous leukemia diagnosed by methods of morphology and cell chemistry. Peripheral blood and bone marrow samples of them were analyzed Philadelphia (Ph1) chromosome by cytogenetic technique. Among them, 5 patients were tested by FISH technique on the slide of interphase and remainders were tested by FISH technique on the slide of metaphase cell. Results: Results of analyzing chromosome of 10 patients showed that 8 patients had Ph1 chromosome. 2 patients without Ph1 chromosome were patients who had not high of leukocyte count: 28x109leukocyte/l and 36x109leukocyte/l, respectively. In the FISH on the slide of interphase, all 5 patients had Ph1 chromosome and abl/bcr fusion gene. In the FISH on the slide of metaphase cell, 3 patients had Ph1 chromosome and abl/bcr fusion gene. Conclusion: FISH technique has been applied successfully to detect ABL/BCR gene in patients with chronic myelogenous leukemia.\r\n', u'\r\n', u'

Identify the \xdf-thalassemia mutation spectrum in the North of Vietnam

Background: \u03b2-thalassemia is a hereditary disease caused by disorder in \u03b2-globin chain synthesis process. In this research, multiplex-PCR was used in combination with blood chemistry assays and clinical symptoms to detect \u03b2-globin mutations. Objectives: (1) to identify the \u03b2-thalassemia mutation spectrum in the North of Vietnam; (2) to determine the relation between biochemistry values and types of mutations. Subject and methods: Blood samples collected from 60 pediatric patients were used in screening assays (hemoglobin counting, red blood cell counting, hematocrit\ufffd? and multiplex-PCR to detect 6 point mutations with high prevalence in the region. Results: \r\n', u'(1) Of 60 blood samples collected from pediatric patients, 30 (50%) had mutation in codon 17 (A\u2192T), 6 (10%) had a frameshift mutation in codons 41/42 and 4 (6%) had both types of these mutations; (2) The average onset time in patients with FS 41/42 mutation was earlier than that of patients with codon 17 (A\u2192T) mutation, whereas transfusion interval did not differ significantly among these patients; (3) Mean corpuscular volume (MCV) was lower in patients with homozygous mutations (\u03b2o) (average 64.8) than in those with heterozygous mutations (\u03b2+) (average 72.7). Conclusions: (1) multiplex-PCR is an effective technique in identifying the mutation spectrum of \u03b2-globin gene in the North of Vietnam; (2) Biochemistry assays should be associated with molecular techniques in diagnose of \u03b2-thalassemia\r\n', u'\r\n', u'\r\n', u'

Using and evaluating cost-effectiveness of NAT in screening HIV, HCV, HBV in blood donors

Background: Nucleic acid testing (NAT) has been widely used for transfusion - transmitted infection screening at blood banks all over the world to reduce window period, yet the assay has not been implemented in Vietnam. Objective: Using and evaluating cost - effectiveness of NAT in screening HIV, HCV, HBV in blood \r\n', u'donors. Subjects and methods: The study was carried out on 9392 blood donors at National Institute of Hematology and Blood Transfusion from Jan to May 2007 who were HIV, HCV, HBV negative with ELISA. Plasma from donors was pooled (pool size of 8) and tested with UItrio Procleix HIV - 1, HCV, HBV (Chiton). Results: These 9392 plasma samples were pooled into 1174 pool samples to perform NAT. Among 1174 pooled samples, there was only 1 case with negative ELISA - Reactive NAT. The sample was determined as response with probe HCV. From there, one of eight pool samples was identified responding to probe HCV and it was more likely to have been missed in the window period when screened by ELISA.Conclusion: The sample should be further tested with HCV qualitative and quantitative testing to confirm the status of infection. \r\n', u'\r\n', u'

Apply Nested-RT-PCR technique to identify bcr/abl fusion gene in the chronic myelogenous leukemia

Background: Bcr/abl fusion gene plays an important role in diagnosing and treating chronic myelogenous leukemia. Objective: to detect fusion genes: b3a2, b2a2, b3a3, b2a3 and e1a2 in patients with chronic myelogenous leukemia by using Nested RT - PCR technique. Subjects and methods: Peripheral blood samples were analyzed by Nested RT - PCR assay from 30 adult patients. Results: 28/30 patients showed bcr/abl fusions gene; among them 20/30 patients showed b3a2 fusions gene, 5/30 patients showed b2a2 fusions gene, 2/30 patients showed co-expression of the b3a2 and b2a2. 1/30 showed e1a2; 2/30 patients showed negative fusion gene. Count of leukocytes and platelets of patients with b3a2 fusion genes were 311.3 G/l and 597.5 G/l, respectively and of patients with b2a2 fusion genes were 136.7 G/l and 333 G/l, respectively. Conclusion: Most of patients showed b3a2 fusion gene, while remaining showed b2a2 transcripts or the co-expression of the b3a2, only one case showed e1a2 fusion gene, two patients showed negative fusion gene. There was no case which showed b3a3 or b2a3 fusion gene. Nested RT assay should be used to determine bcr/abl fusion genes for patients with chronic myelogenous leukemia\r\n', u'\r\n', u'