Researchers from the MLL Munich Leukaemia Laboratory have used 454 Sequencing methods to distinguish and characterise forms of leukaemia and myeloproliferative disorders.
The researchers presented the results at the 51st American Society for Hematology Annual Meeting.
Using targeted resequencing techniques from Roche Applied Science, including Nimblegen Sequence Capture arrays and 454 Life Science's GS FLX System, the researchers were able to successfully detect all types of molecular mutations identified by conventional methods and, in addition, identify novel mutations in leukaemia samples.
Importantly, the researchers were able to characterise a range of genetic variation types, such as point mutations, insertions and deletions, as well as chromosomal rearrangements, in a single sequencing run; current methods require a combination of different labour-intensive techniques, including FISH and standard Sanger sequencing.
The results have critical implications on research to develop future diagnostics assays and treatments for this disease.
Leukaemia is a cancer of the blood that causes rapid, abnormal proliferation of blood cells and consists of a broad spectrum of subtypes.
While a number of treatment options are available, understanding the genetics and molecular composition of an individual's leukaemia type is essential to determining the best course of action.
Current methodologies are labour-intensive, expensive, rely on expert knowledge and often lack the sensitivity required to detect rare mutations.
The MLL Munich Leukaemia Laboratory team, led by Dr Torsten Haferlach, recognised the power and speed of high-throughput sequencing to address these issues.
Dr Haferlach said: 'We identified 454 Sequencing technology as a promising method to characterise leukaemia and other haematological malignancies.
'In our research on a variety of leukaemia types and myeloproliferative neoplasms, we confirmed that not only are we able to comprehensively detect all types of known molecular mutations, but to identify also novel mutations, such as fusion partner genes resulting from balanced translocation events,' he added.
In one such study, the researchers used Nimblegen Sequence Capture 385K arrays to enrich a 1.91Mb region of the genome containing 95 cancer-associated genes in six acute myeloid leukaemia (AML) samples.
They then sequenced the captured DNA with the GS FLX Titanium series chemistry and analysed the results with the company's GS Reference Mapper software.
The results showed for the first time that point mutations, deletions and insertions, as well as fusion genes from translocations and inversions, could be detected in a one-step methodological approach.
Another study - presented by Dr Alexander Kohlmann, PhD and head of the NGS group at the MLL, and Vera Grossmann - used ultra-deep sequencing of amplicons to accurately identify mutations in oncogenic regions within 95 samples of leukaemia and myeloproliferative neoplasms.
Dr Kohlmann said: 'Amplicon sequencing with the GS FLX System is a particularly straightforward and powerful method to detect a wide range of molecular mutations with high sensitivity.
'It is of particular utility for characterising the constantly growing number of target genes used to distinguish molecular subtypes of hematological malignancies.
'This technology has the potential to immediately change the way we obtain novel molecular insights underlying this disease,' he added.
