A rapid method for simultaneous multi-gene mutation screening in children with nonsyndromic hearing loss

Abstract

Hearing loss (HL) is a common genetically heterogeneous sensory disorder, occurring in 1 to 3 per 1000 live births. In spite of the extraordinary genetic heterogeneity, variants in GJB2, MT-RNR1, and SLC26A4 genes have been considered as the main reasons of nonsyndromic hearing loss in Chinese population. We developed a rapid multiplex genetic screening system called the SNPscan assay technique which could detect the 115 mutations of the above three genes. This technique is a high-throughput and cost-saving SNP genotyping method. We found that the carrier rate of mutations in the GJB2 gene, MT-RNR1 gene, and SLC26A4 gene was 26.21%, 1.86%, and 25.46% of the patients with nonsyndromic hearing loss, respectively. Using this method, up to 50% of the patients in our study were identified to have hereditary HL caused by mutations in the three genes. It is applicable to not only genetic diagnosis of HL, but also molecular screening of other inherited diseases.

SNPscan as a high-performance screening tool for mutation hotspots of hearing loss-associated genes

“…The SNPscan technology, a recently developed multiplex genotyping method, is an alternative method that overcomes the limitation of SNaPshot. SNPscan was used for the first time by Du et al in mutation screening in subjects with HL; the results demonstrated that SNPscan assay is a robust tool for studies on genetic NSHL [14]. To further investigate the feasibility of this method in clinical testing, we compared our 15 loci SNaPshot analysis system with the newly developed 115 loci SNPscan in 162 children with HL and 276 normal children.…”

“…To overcome the limitations of traditional detection methods, we introduce a newly developed multiplex genetic screening system called the SNPscan technique to investigate neonatal HL in Chinese population. This screening system can simultaneously detect 115 mutations in the GJB2, SLC26A4, and MT-RNR1 genes, which covered the majority of mutations associated with genetic HL in Chinese population [14]. In this study, we compared the SNPscan technique with the SNaPshot screening system described in our previous study and discussed the feasibility of the SNPscan technique for mutation screening in neonatal HL [15].…”

Heterozygote Advantage of the rs3794624 Polymorphism in CYBA for Resistance to Tuberculosis in Two Chinese Populations

“…Genotyping of the discovery cohort was carried out by Sequenom’s iPLEX SNP genotyping protocol using matrix-assisted laser desorption/ionization time of flight mass spectrometry on the MassArray Analyzer 4 system (Sequenom Inc., San Diego, CA, USA). SNP genotyping of the replication cohort was performed using a SNPscan Kit (Cat#:G0104, Genesky Biotechnologies Inc., Shanghai, China) as described previously25. The SNPscan genotyping technology is based on double ligation and multiplex fluorescence polymerase chain reactions.…”

Association of the VRK2 gene rs3732136 polymorphism with schizophrenia in a Northwest Chinese Han population

“…A 48-Plex SNPscan TM kit (Genesky Biotechnologies Inc., Shanghai, China) was designed and used to determine genotypes of the four SNPs. The SNPscan technique was developed according to patented SNP genotyping technology by Genesky Biotechnologies Inc., which provides a high-throughput and cost-saving SNP genotyping method based on double ligation and multiplex fluorescence PCR as previously described (Du et al, 2014).…”