Oligonucleotide microarray based method for simultaneous diagnostic of 3-M syndrome, SOPH syndrome, tyrosinemia type 1, methaemoglobinaemia type 1, nonsyndromic hearing loss and deafness (DFNB1)

Background and objectives. High incidence and high prevalence of hereditary diseases in Yakut ethnic group of Republic of Sakha (Yakutia) has been reported by several research groups. The incidence of some of the diseases are much higher comparing to data in other regions of Russia and the worldwide. There is a pressing need for prevention of those diseases through the mass genetic screening of population. The methods of genetic diagnostics that are known today are not able to afford the mass screening due to the high cost and time consumption for a single test. One of promising technologies and methods of DNA diagnostics is DNA microarray. It enables researchers to quickly screen large numbers of biological analytes for a variety of purposes including disease diagnostics. Methods. A Perkin Elmer Piezzorray (Perkin Elmer) microarrayer was used to print oligonucleotide based low-density microarrays by non-contact manner for addressing 5 mutations causing 5 frequently occurring diseases reported from yakut population. The assay using developed microarray chip based on reverse hybridization which include two step multiplex PCR reactions with production of one-stranded Cy5 labeled-PCR products following its hybridization with oligonucleotide probes. Results. In this study we developed a DNA microarray for detection of mutations in genes CUL7, NBAS, FAH, DIA1, GJB2 genes which are known to be cause of five very frequently occurring hereditary diseases in republic of Sakha (Yakutia): 3-M syndrome, SOPH-syndrome, tyrosinemia type 1, methaemoglobinaemia type 1, nonsyndromic hearing loss and deafness (DFNB1) type 1A respectively. Testing of experimental versions of DNA chips as well as control DNA diagnostic of five hereditary diseases on the same DNA samples using PCR with PCR and restriction enzyme analysis and real-time PCR were carried out.

наследственные заболевания, якуты, точковые мутации, гетерозиготное носительство, диагностический биочип, синтетические олигонуклеотиды, 3-М синдром, SOPH-синдром, тирозинемия 1 типа, метгемоглобинемия 1 типа, несиндромальная глухота 1 типа, inherited diseases, Yakuts, point mutatios, heterozygous carriage, DNA-microarray, diagnostics, synthesized oligonucleotides, 3-M syndrome, SOPH syndrome, tyrosinemia type 1, methaemoglobinaemia type 1, nonsyndromic hearing loss and deafness (DFNB1) type 1A