MICROARRAY MEETING 2002: NEW DEVELOPMENTS IN MUTATION DETECTION AND GENE EXPRESSION
Segrate, MI (Italy), April 12, 2002 

Minerva Biotecnologica 2002 December;14(3-4):241-6

Copyright © 2003 EDIZIONI MINERVA MEDICA

lingua: Inglese

Single nucleotide polymorphism and mutation identification by microelectronic chip technology

Stenirri S. ¹, Foglieni B. ¹, Manitto M. P. ², Martina E. ², Brancato R. ², Cremonesi L. ¹, Ferrari M. ^{1, 3}

¹ Unit of Genomics for Diagnosis of Human Pathologies, IRCCS H. San Raffaele, Milan, Italy; ² Department of Ophthalmology and Visual Sciences, University Hospital San Raffaele, Milan, Italy; ³ Diagnostica e Ricerca, San Raffaele SpA, Milan, Italy

Back­ground. The ­need of devel­op­ing ­advanced meth­o­dol­o­gies for muta­tion iden­tifi­ca­tion has ­been rais­ing in the ­last few ­years. ­This is prin­ci­pal­ly due to the iden­tifi­ca­tion of an increas­ing num­ber of genet­ic vari­a­tions asso­ciat­ed ­with ­human dis­eas­es. Micro­ar­rays are a mod­ern DNA diag­nos­tic ­tool, allow­ing the rap­id anal­y­sis of a ­large ­cohort of ­patients for a ­huge num­ber of alle­lic var­i­ants.
Meth­ods. In the ­present ­study we ­have ­employed a new micro­elec­tron­ic ­based tech­nol­o­gy: the NMW 1000 Nano­Chip™ Molec­u­lar Biol­o­gy Work­sta­tion (Nano­gen, San Die­go, CA), ­that ­enables the ­active move­ment of ­charged mole­cules to des­ig­nat­ed ­test ­sites. We devel­oped ­assays for the iden­tifi­ca­tion of ­some com­mon Ital­ian muta­tions in the ret­i­na-spe­cif­ic ABC trans­port­er (­ABCA4) ­gene, ­involved in Star­gardt dis­ease. ­Assays ­were val­i­dat­ed by a ret­ro­spec­tive ­study on a ­large num­ber of ­wild ­type and mutat­ed sam­ples.
­Results. Com­par­i­son of the ­results ­obtained ­with the Nano­gen tech­nol­o­gy and ­those ­obtained ­with oth­er meth­ods ­showed a com­plete con­cor­dance.
Con­clu­sions. In our expe­ri­ence, the micro­elec­tron­ic tech­nol­o­gy devel­oped by Nano­gen ­allowed to over­come ­some of the lim­i­ta­tions due to pas­sive hybrid­iza­tion. ­This, cou­pled to the pos­sibil­ity of assem­bling dif­fer­ent com­bi­na­tions of spe­cif­ic ­probes ­onto a micro­chip to per­form cus­to­mized anal­y­ses, ­could ­improve the effi­cien­cy of muta­tion iden­tifi­ca­tion in a varie­ty of genet­ic diag­nos­tic appli­ca­tions.