Rapid microsatellite analysis of paraffin embedded tumour specimens from patients with hereditary non-polyposis colorectal cancer.
In screening for hereditary non-polyposis colorectal cancer (HNPCC)--an autosomal dominant disorder characterised by mutations in mismatch repair genes--detection of microsatellite instability is an important diagnostic criterion. The mono- or dinucleotide repeat DNA sequences are usually amplified from formalin fixed, paraffin embedded tissue by polymerase chain reaction after numerous time consuming steps including deparaffinisation, DNA extraction, and purification. A rapid single step method for direct DNA analysis is described, based on preincubation of paraffin embedded tissue with Triton X-100 followed by DNA amplification with fluorescence labelled primers and electrophoresis in an automated sequencer. This procedure allows precise allele sizing and analysis of genetic instability, is more efficient and time saving, reduces the risk of contamination, and is therefore of particular interest in screening for HNPCC.
Raedle
J
J
Johann Wolfgang Goethe-University, Frankfurt/Main, Germany.
Brieger
A
A
Trojan
J
J
Herrmann
G
G
Zeuzem
S
S
D003123:Colorectal Neoplasms, Hereditary Nonpolyposis / Q000175:diagnosis* / Q000235:genetics; D004273:DNA, Neoplasm / Q000032:analysis; D005820:Genetic Testing / Q000379:methods; D006801:Humans; D018895:Microsatellite Repeats; D016612:Paraffin Embedding; D016133:Polymerase Chain Reaction / Q000379:methods; D013997:Time Factors
D004273:DNA, Neoplasm