In Situ Hybridization Methods to Study Microbial Populations and Their Interactions with Human Host Cells

2004 ◽  
pp. 1-25 ◽  
Author(s):  
Ruth Holm
Keyword(s):  
In Situ Hybridization ◽  
Host Cells ◽  
Microbial Populations ◽  
Human Host

Fluorescence In Situ Hybridization Detected MRD and Chimerism in Patients with Hematologic Malignancies after allo-HSCT.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 5214-5214
Author(s):  
Huiying Qiu ◽  
Yongquan Xue ◽  
Jinlan Jin ◽  
De Pei Wu
Keyword(s):  
Stem Cell ◽  
In Situ Hybridization ◽  
Stem Cell Transplantation ◽  
Fluorescence In Situ Hybridization ◽  
Cell Transplantation ◽  
Residual Disease ◽  
Hematologic Malignancies ◽  
Host Cells ◽  
Hematopoietic Stem

Abstract Objective Monitoring of minimal residual disease (MRD) and cellular chimerism in patients with hematopoietic malignancies after allogeneic hematopoietic stem cell transplantation(allo-HSCT). Methods From May 2001 to June 2005, seventy four patients were received allo-HSCT. Including 50 Males and 24 females. 41 patients received sibling HLA-matched BMT, 7 patients received un-related BMT, 9 patients received Nonmyeloablative stem cell transplantation (NST), 14 patients received related haploidentical transplantation and 3 patients received allo-PBSCT. Among them, 45 patients were diagnosed with CML, 13 patients with AML, 14 patients with ALL, one patient with Multiple myeloma and one patient with malignant lymphoma. Chimerism and MRD were monitored using X and Y specific centromeric probes or gene probes for BCR/ABL, MLL and AML1/ETO by fluorescence in situ hybridization (FISH),1000 cells were analysised. Results Among 18 patients, received sex-matched transplant, we did not found the former chromosome rearrangements in 14 patients after transplantation, MRD were detected in 17% and 10% of cells in two patients, MRD were decreased from 10% to 1% of cells after the reduction of the dose of immunotherapy in one patient four month later, the patient was still in remission one year after transplantation. Another patient died of sever GVHD after the reduction of immunotherapy. 2 patients were found to have the former chromosomal rearrangement 1 and 4 month after transplantation respectively who did not achieve remission after chemotherapy and died 3 and 5 months respectively after transplantation. Over 99% donor chimerisms were found in 44 patients on day 25, donor cells were at a low level (96.2%~98.7) in 7 patients on day 25, and increased over 99% on day 180, they were in remission without relapse. The donor chimerisms decreased gradually in 6 patients, host cells were found over 20 cells, 3 patients showed cytogenetic or hematologic bone marrow relapse, two patients died of sever GVHD after the reduction of cyclosporine A, Over 99% donor chimerisms was achieved in one patient. Conclusion FISH could play a pivotal role in the detection of MRD and chimerism. It is helpful to the evaluation of graft and relapse, to the guide of implement of early immunotherapy.

Analysis of the origin of marrow cells in bone marrow transplant recipients using a Y-chromosome-specific in situ hybridization assay

Blood ◽  
1989 ◽  
Vol 74 (6) ◽  
pp. 2220-2226 ◽  
Author(s):  
DM Durnam ◽  
KR Anders ◽  
L Fisher ◽  
J O'Quigley ◽  
EM Bryant ◽  
...  
Keyword(s):  
Bone Marrow ◽  
In Situ Hybridization ◽  
Y Chromosome ◽  
Bone Marrow Cells ◽  
Conditioning Regimen ◽  
Disease Status ◽  
Host Cells ◽  
Hybridization Assay ◽  
Marrow Cells

A Y-chromosome-specific in situ hybridization assay was used to assess the frequency with which host bone marrow cells are retained after marrow grafting. The majority of patients (74%) showed the presence of both host and donor marrow cells when assayed 14 days after transplant. By 84 days posttransplant only 4% of the patients retained host marrow cells. Only 1 of 19 evaluable patients analyzed over 1 year posttransplant showed minimal retention of host cells. No statistical correlation was found between retention of host cells posttransplant and the development of relapse or acute or chronic graft-versus-host disease. Pretransplant conditioning regimen, HLA-matching, diagnosis, disease status at transplant, ABO-matching, and patient age also showed no correlation with the retention of host cells posttransplant.

Analysis of the origin of marrow cells in bone marrow transplant recipients using a Y-chromosome-specific in situ hybridization assay

Blood ◽  
1989 ◽  
Vol 74 (6) ◽  
pp. 2220-2226 ◽  
Author(s):  
DM Durnam ◽  
KR Anders ◽  
L Fisher ◽  
J O'Quigley ◽  
EM Bryant ◽  
...  
Keyword(s):  
Bone Marrow ◽  
In Situ Hybridization ◽  
Y Chromosome ◽  
Bone Marrow Cells ◽  
Conditioning Regimen ◽  
Disease Status ◽  
Host Cells ◽  
Hybridization Assay ◽  
Marrow Cells

Abstract A Y-chromosome-specific in situ hybridization assay was used to assess the frequency with which host bone marrow cells are retained after marrow grafting. The majority of patients (74%) showed the presence of both host and donor marrow cells when assayed 14 days after transplant. By 84 days posttransplant only 4% of the patients retained host marrow cells. Only 1 of 19 evaluable patients analyzed over 1 year posttransplant showed minimal retention of host cells. No statistical correlation was found between retention of host cells posttransplant and the development of relapse or acute or chronic graft-versus-host disease. Pretransplant conditioning regimen, HLA-matching, diagnosis, disease status at transplant, ABO-matching, and patient age also showed no correlation with the retention of host cells posttransplant.

scholarly journals Fixation-free fluorescence in situ hybridization for targeted enrichment of microbial populations

2010 ◽  
Vol 4 (10) ◽  
pp. 1352-1356 ◽  
Author(s):  
Suzan Yilmaz ◽  
Mohamed F Haroon ◽  
Brian A Rabkin ◽  
Gene W Tyson ◽  
Philip Hugenholtz
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Microbial Populations ◽  
Targeted Enrichment

The Use of CARD-FISH to Evaluate the Quantitative Microbial Ecology Involved in the Continuous Bioleaching of a Cobaltiferrous Pyrite

2007 ◽  
Vol 20-21 ◽  
pp. 565-568 ◽  
Author(s):  
E. Díaz ◽  
E. González-Toril ◽  
Catherine Joulian ◽  
R. Amils
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Microbial Populations ◽  
Powerful Method ◽  
High Fluorescence ◽  
Catalyzed Reporter Deposition ◽  
Card Fish ◽  
Slow Growing ◽  
Signal Intensities

Catalyzed reporter deposition fluorescence in situ hybridization (CARD-FISH) is a powerful method with a growing number of applications in the quantitative evaluation of microbial populations of complex ecosystems. CARD-FISH is an improvement over traditional fluorescence in situ hybridization (FISH) especially suitable for aquatic habitats with small, slow growing, or starving bacteria, in which the signal intensities of hybridized cells is frequently below detection limits or lost in high fluorescence background of dense mineral matrixes. In this work we report the development of protocols and probes for the identification and quantification of the microorganisms involved in the continuous bioleaching of a cobaltiferrous concentrate using a four tank-leaching reactor operated by BRGM. After steady state was reached, samples were taken to identify and quantify the microorganisms present in the each of the tanks used in the process.

DNA sequence mapping in interphase and metaphase chromosomes by fluorescence in situ hybridization

1992 ◽  
Vol 50 (1) ◽  
pp. 496-497
Author(s):  
Barbara Trask ◽  
Susan Allen ◽  
Anne Bergmann ◽  
Mari Christensen ◽  
Anne Fertitta ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Dna Sequence ◽  
Dna Sequences ◽  
Dual Band ◽  
Nick Translation ◽  
Metaphase Chromosomes ◽  
Band Pass ◽  
Texas Red ◽  
Fluorescent Spot

Using fluorescence in situ hybridization (FISH), the positions of DNA sequences can be discretely marked with a fluorescent spot. The efficiency of marking DNA sequences of the size cloned in cosmids is 90-95%, and the fluorescent spots produced after FISH are ≈0.3 μm in diameter. Sites of two sequences can be distinguished using two-color FISH. Different reporter molecules, such as biotin or digoxigenin, are incorporated into DNA sequence probes by nick translation. These reporter molecules are labeled after hybridization with different fluorochromes, e.g., FITC and Texas Red. The development of dual band pass filters (Chromatechnology) allows these fluorochromes to be photographed simultaneously without registration shift.

Ultrastructural analysis of the spatial distribution of MRNA

1992 ◽  
Vol 50 (1) ◽  
pp. 552-553
Author(s):  
Gary Bassell ◽  
Robert H. Singer
Keyword(s):  
Electron Microscopy ◽  
Spatial Distribution ◽  
In Situ Hybridization ◽  
High Resolution ◽  
Nucleic Acids ◽  
Colloidal Gold ◽  
Dna Probe ◽  
Nucleotide Analogs ◽  
Gold Particles

We have been investigating the spatial distribution of nucleic acids intracellularly using in situ hybridization. The use of non-isotopic nucleotide analogs incorporated into the DNA probe allows the detection of the probe at its site of hybridization within the cell. This approach therefore is compatible with the high resolution available by electron microscopy. Biotinated or digoxigenated probe can be detected by antibodies conjugated to colloidal gold. Because mRNA serves as a template for the probe fragments, the colloidal gold particles are detected as arrays which allow it to be unequivocally distinguished from background.

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