Multiplex tetra-primer amplification refractory mutation system PCR to detect 6 common germline mutations of the MUTYH gene associated with polyposis and colorectal cancer.
BACKGROUND
We describe a simple tetra-primer amplification refractory mutation system PCR (T-ARMS-PCR) for detecting MUTYH mutations, which are associated with colorectal adenomas and colorectal cancer.


METHODS
We designed specific T-ARMS-PCR assays for 6 mutations (Y165C, G382D, 1395_7delGGA, Y90X, 1103delC, and R231H) selected on the basis of the frequency of their occurrence. We also designed a set of 3 multiplex T-ARMS PCR assays, each for detection of 2 mutations. We tested DNA samples from patients with attenuated or classic adenomatous polyposis coli and no detectable APC germline mutations.


RESULTS
All mutations were easily detected with both the specific and multiplex T-ARMS-PCR assays. Results were confirmed by DNA HPLC analysis in all 54 patients, and each mutation was confirmed by direct DNA sequencing.


CONCLUSIONS
T-ARMS-PCR does not require any special equipment, and it provides rapid, reproducible, and cost-effective detection of common MUTYH mutations. Multiplex T-ARMS-PCR allows the detection of 6 common MUTYH mutations with use of as few as 3 single tube PCR reactions. It could be useful to carry out large population-based epidemiologic studies.
Piccioli
Patrizia
P
Laboratory of Human Genetics, Medical Oncology C, Istituto Nazionale per la Ricerca sul Cancro, Genoa, Italy.
Serra
Martina
M
Gismondi
Viviana
V
Pedemonte
Simona
S
Loiacono
Fabrizio
F
Lastraioli
Sonia
S
Bertario
Lucio
L
De Angioletti
Maria
M
Varesco
Liliana
L
Notaro
Rosario
R
D011125:Adenomatous Polyposis Coli / Q000235:genetics*; D015179:Colorectal Neoplasms / Q000235:genetics*; D045647:DNA Glycosylases / Q000235:genetics*; D018095:Germ-Line Mutation; D006801:Humans; D016133:Polymerase Chain Reaction
D045647:DNA Glycosylases; C061738:mutY adenine glycosylase