Monoquant Pty Ltd.

Other

The Trustees of Flinders University

Alexander Alan Morley

Michael Julian Brisco

Michael J Brisco

Pamela Sykes

Michael Brisco

Michel Julian Brisco

Aspects of the present invention relate to methods of assessing the amplification efficiency of a primer especially the relative or absolute efficiency of a primer used in quantitative PCR amplification. Some of the methods described herein involve the assessment of primer efficiency relative to the Ct determined for a single target DNA molecule rather than Ct regression curve analysis of serially diluted samples. By assessing primer efficiency utilizing the approaches described herein, one can more efficiently select primers for the diagnosis and/or monitoring of disease conditions, analysis of specific gene regions of interest, the monitoring of disease conditions that are characterised by clonal lymphoid cell populations or disease conditions that are characterised by specific V(D)J recombination events including the detection of minimal or residual disease in leukaemia patients.

Method of assessment

The present invention relates generally to an improved method of amplifying a nucleic acid region of interest and to primers for use therein. More particularly, the present invention is directed to an improved method of amplifying a nucleic acid region which has resulted from the recombination of two or more immunoglobulin or T cell receptor gene segments and primers for use therein. The method of the present invention is based on the determination that performing the amplification step at an annealing temperature determined relative to the critical annealing temperature unique to a given reaction and/or using optimised primers enables higher levels of sensitivity than have previously been achievable in the context of prior art methods of amplifying rearranged immunological or T cell receptor genes. The method of the present invention is particularly useful where the subject recombination target comprises only one N region. The provision of a highly sensitive yet simple means of detecting specific immunological and T cell receptor nucleic acid recombination events is useful in a range of applications including, but not limited to, the diagnosis and/or monitoring of clonal lymphoid cell populations or disease conditions which are characterised by specific V/D/J recombination events (such as detecting minimal residual disease in leukaemias) or the analysis or identification of immunological or T cell receptor gene regions of interest.

Nucleic acid amplification method and primers for use therein

A method of determining a quantitative measure of the integrity of RNA in a sample, the method comprising: (i) assaying a sample containing instances of an RNA molecule transcribed from a reference gene, at least some of the instances being damaged, to determine quantitative measures of the relative or absolute numbers of intact instances of each of a plurality of segments of the RNA molecule in the sample, the segments having respective different lengths; (ii) based on a relationship between the determined quantitative measures and the respective different lengths of the segments, determining a quantitative measure of integrity of the instances of the RNA molecule in the sample; and (iii) determining the total number of instances of an RNA molecule of interest in a sample by using the quantitative measure of integrity and the length of a corresponding degradation-relevant segment of the RNA molecule of interest.

Method of determining RNA integrity

The present invention relates to a method for identifying a DNA breakpoint and agents for use therein. More particularly, the present invention provides a method for identifying a gene translocation breakpoint based on the application of a novel multiplex DNA amplification technique. The method of the present invention facilitates not only the identification of the breakpoint position but, further, enables the isolation of the DNA segment across which the breakpoint occurs. This provides a valuable opportunity to conduct further analysis of the breakpoint region, such as to sequence across this region. The method of the present invention is useful in a range of applications including, but not limited to, providing a routine means to characterize the gene breakpoint associated with disease onset in a patient and thereby enable the design of patient specific probes and primers for ongoing monitoring of the subject disease condition. In addition to monitoring the progression of a condition characterized by the existence of the breakpoint, there is also enabled assessment of the effectiveness of existing therapeutic drugs and/or new therapeutic drugs and, to the extent that the condition is a neoplasm, prediction of the likelihood of a subject's relapse from a remissive state.

Method for DNA breakpoint analysis

The present invention relates generally to a method of amplifying a nucleic acid region of interest and, more particularly, to a method of amplifying a nucleic acid region of interest via a nested single tube PCR. The method is designed to provide a means to selectively inactivate the functionality of the outer primer or primers and to maintain amplification efficiency throughout the reaction. The development of a means to achieve efficient amplification by the outer primer followed by efficient amplification with the inner primers, in the context of a single tube nested PCR, is useful in a range of applications including, but not limited to, the diagnosis and/or monitoring of disease conditions which are characterized by specific gene sequences and the characterization or analysis of specific gene regions of interest. Still further, the method enables quantification to be performed and not just simple detection.

Method of amplification

The present invention relates to a method of characterizing a nucleic acid region and, more particularly, to a method of analysing a marker nucleic acid region. The method of the present invention is based on identification of one or both of the nucleic acid regions flanking a marker nucleic acid region and provides a means of analysing a marker nucleic acid region which is characteristic of a clonal population of cells. The method of the present invention is useful in the context of enabling a range of applications including, but not limited to, monitoring the progression of a condition characterized by the presence of a clonal populations of cells (such as a neoplastic condition), monitoring the levels of one or more clonal cell population, predicting the likelihood of a subject's relapse from a remissive state to a disease state, for assessing the effectiveness of existing therapeutic drugs and/or new therapeutic agents and identifying the presence of a marker region.

Method of analysing a marker nucleic acid molecule

The present invention is directed to a method for the detection of leukemia or lymphoma by determining the homogeneity or heterogeneity of the length of immunoglobulin or T-receptor gene segments. The present invention is further directed to a kit that is useful for the detection of leukemia or lymphoma.

Growing community of inventors

Glenelg, Australia

Alexander Alan Morley