Sequence-specific Oligonucleotide Probe Polymerase Chain Reaction

2020 ◽  
Author(s):  
Blood ◽  
1993 ◽  
Vol 81 (2) ◽  
pp. 551-559 ◽  
Author(s):  
WR Drobyski ◽  
LA Baxter-Lowe ◽  
RL Truitt

Disease relapse after allogeneic bone marrow transplantation (BMT) is a major cause of treatment failure and is thought to evolve from clinically occult residual disease in the recipient. However, the demonstration of minimal residual disease (MRD) in individual patients is of uncertain prognostic significance because the detection of residual disease has not consistently correlated with subsequent relapse. Moreover, the optimal therapeutic approach in patients with MRD after allogeneic BMT is unknown. The study of these issues has been hindered by the lack of clinically relevant animal models. In this report, we characterize a novel murine model for the study of MRD after allogeneic BMT. This model was designed to simulate high-risk BMT in humans in which patients receive transplants in relapse and disease recurrence in the major cause of treatment failure. The H-2-compatible, mixed lymphocyte culture nonreactive murine strains, AKR (H-2k) and CBA (H-2k), were chosen to parallel marrow transplants from HLA-matched siblings, which represent the majority of allo-transplants in humans. Male AKR leukemia cells were used in female donor/host chimeras permitting the Y chromosome to serve as a leukemia-specific marker for MRD. Detection of residual male leukemia cells in the peripheral blood of the primary host was facilitated by use of the polymerase chain reaction (PCR) and sequence-specific oligonucleotide probe hybridization (SSOPH). Use of PCR/SSOPH was highly predictive of clinical outcome (relapse or cure) in animals receiving transplants (P < .00002) and detected disease recurrence earlier than comparative flow cytometric analysis studies. This murine model will be useful in evaluating the efficacy of therapeutic strategies aimed at reducing disease relapse posttransplant and can be adapted to other transplant murine tumor systems for the study of MRD.


Blood ◽  
1993 ◽  
Vol 81 (2) ◽  
pp. 551-559 ◽  
Author(s):  
WR Drobyski ◽  
LA Baxter-Lowe ◽  
RL Truitt

Abstract Disease relapse after allogeneic bone marrow transplantation (BMT) is a major cause of treatment failure and is thought to evolve from clinically occult residual disease in the recipient. However, the demonstration of minimal residual disease (MRD) in individual patients is of uncertain prognostic significance because the detection of residual disease has not consistently correlated with subsequent relapse. Moreover, the optimal therapeutic approach in patients with MRD after allogeneic BMT is unknown. The study of these issues has been hindered by the lack of clinically relevant animal models. In this report, we characterize a novel murine model for the study of MRD after allogeneic BMT. This model was designed to simulate high-risk BMT in humans in which patients receive transplants in relapse and disease recurrence in the major cause of treatment failure. The H-2-compatible, mixed lymphocyte culture nonreactive murine strains, AKR (H-2k) and CBA (H-2k), were chosen to parallel marrow transplants from HLA-matched siblings, which represent the majority of allo-transplants in humans. Male AKR leukemia cells were used in female donor/host chimeras permitting the Y chromosome to serve as a leukemia-specific marker for MRD. Detection of residual male leukemia cells in the peripheral blood of the primary host was facilitated by use of the polymerase chain reaction (PCR) and sequence-specific oligonucleotide probe hybridization (SSOPH). Use of PCR/SSOPH was highly predictive of clinical outcome (relapse or cure) in animals receiving transplants (P < .00002) and detected disease recurrence earlier than comparative flow cytometric analysis studies. This murine model will be useful in evaluating the efficacy of therapeutic strategies aimed at reducing disease relapse posttransplant and can be adapted to other transplant murine tumor systems for the study of MRD.


1994 ◽  
Vol 112 (1) ◽  
pp. 33-43 ◽  
Author(s):  
L. Rexach ◽  
F. Dilasser ◽  
P. Fach

SummaryThe important role of plasmid genes in assessing virulence for BALB/c mice in salmonella, and the difficulty of using standard techniques to detect them, led us to develop a detection method by gene amplification.One hundred and forty–three strains (71 serovars) of salmonella and 35 strains of other species were tested using specific oligonucleotide primers. The amplification products were identified by a specific oligonucleotide probe. Forty-nine salmonella strains from ten serovars (S. abortus ovis, S. choleraesuis, S. dublin, S. enteritidis, S. gallinarum / pullorum, S. hessarek, S. typhimurium, S. IIIa48:z4, z23,S. IV43: z4, z23:-,S. V28:a:-) produced a positive and specific response.Because of various origins of the strains possessing the gene sought and the diversity of the responses, both from one serovar to another and in the same serovar, this search has its place among the epidemiological markers in general use. This method appears well suited to the research and detection of plasmid genes associated with mouse virulence in salmonella.


1992 ◽  
Vol 95 (1) ◽  
pp. 63-69 ◽  
Author(s):  
Kathleen A. Grant ◽  
Joanne H. Dickinson ◽  
Matthew D. Collins ◽  
Rohan G. Kroll

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