Abstract: 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.

Abstract: 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.

Abstract: 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.

Abstract: 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 using a PCR method designed to minimise the generation of amplicons from primers which have bound to nucleic acid regions other than the specific region of interest. The method of the present invention is based on the determination that by rendering inefficient the functionality of either the forward primer or the reverse primer, the rate of amplification of irrelevant nucleic acid regions can be reduced relative to amplification of the region of interest.

Abstract: 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.

Abstract: 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 characterise 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.

Abstract: The present invention relates to a method of characterising 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 characterised 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.