Detection of APC mutation by real-time polymerase chain reaction and melting curve analysis / 中华病理学杂志
Chinese Journal of Pathology
;
(12): 669-671, 2005.
Article
in Chinese
| WPRIM
| ID: wpr-258301
ABSTRACT
<p><b>OBJECTIVE</b>To establish a simple, reliable and low cost approach for clinical detection of APC mutation.</p><p><b>METHODS</b>Using SYBR Green I as the real-time polymerase chain reaction (PCR) product indicator, a DNA fragment of 270 bp targeting APC_1309 mutation (5 bp deletion) was amplified from the sample DNA. A short fragment (40/35 bp) was then amplified from the 270 bp PCR product, followed by melting curve analysis from 65 degrees C to 99 degrees C at 0.5 degrees C/step.</p><p><b>RESULTS</b>A total of 18 paraffin-embedded tumor samples were analyzed, of which 7 were tested positive for the mutation and 11 were negative. No mutation was detected in any of the 20 normal peripheral blood samples.</p><p><b>CONCLUSIONS</b>Real-time PCR melting curve analysis can be used for routine APC mutation detection. The simple design, low cost and high reliability should allow similar applications to the analysis of a variety of other gene mutations.</p>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
DNA, Neoplasm
/
Base Sequence
/
Colorectal Neoplasms
/
Chemistry
/
Polymerase Chain Reaction
/
Sequence Deletion
/
Genes, APC
/
Adenomatous Polyposis Coli Protein
/
Genetics
/
Methods
Type of study:
Diagnostic study
Limits:
Humans
Language:
Chinese
Journal:
Chinese Journal of Pathology
Year:
2005
Type:
Article
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