Nucleic acid amplification method and primers for use therein
AssigneeMonoquant Pty Ltd.
InventorAlexander Alan Morley
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AssigneeMonoquant Pty Ltd.
InventorAlexander Alan Morley
Estimating the term…
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.
A method of amplifying an Ig or TCR nucleic acid region which is characterised by the rearrangement of two or more V, D or J gene segments said method comprising contacting a nucleic acid sample of interest with forward and reverse primers directed to said rearranged Ig or TCR nucleic acid region and amplifying said nucleic acid sample using an annealing temperature within the range between Tc and (Tc-3° C.) of the subject amplification reaction, and optionally:
A method of detecting and/or monitoring a clonal population of cells in a mammal, which clonal cells are characterised by an Ig or TCR nucleic acid region which is characterised by the rearrangement of two or more V, D or J gene segments, said method comprising:
Dependencies are inferred from the claim text.