Evaluation of amplification refractory mutation system (ARMS) technique for quick and accurate prenatal gene diagnosis of <em>CHM</em> variant in choroideremia

Abstract

Choroideremia is a rare X-linked recessive inherited disorder that causes chorioretinal dystrophy leading to visual impairment in its early stages which finally causes total blindness in the affected person. It is caused due to mutations in the CHM gene. In this study, we have recruited a pedigree with choroideremia and detected a nonsense variant (c.C799T:p.R267X) in CHM of the proband (I:1). Different primer sets for amplification refractory mutation system (ARMS) were designed and PCR conditions were optimized. Then, we evaluated the sequence variant in the patient, carrier, and a fetus by using ARMS technique to identify if they inherited the pathogenic gene from parental generation; we used amniotic fluid DNA for the diagnosis of the gene in the fetus. The primer pairs, WT2+C and MT+C, amplified high specific products in different DNAs which were verified by Sanger sequencing. Based on our results, ARMS technique is fast, accurate, and reliable prenatal gene diagnostic tool to assess CHM variants. Taken together, our study indicates that ARMS technique can be used as a potential molecular tool in the diagnosis of prenatal mutation for choroideremia as well as other genetic diseases in undeveloped and developing countries, where there might be shortage of medical resources and supplies.

Novel splicing variant c. 208+2T&gt;C inBBS5segregates with Bardet–Biedl syndrome in an Iranian family by targeted exome sequencing

“…Regarding this report and previous studies, essentially retinopathological investigations were driven by mutational data analysis from new molecular based diagnosis, such as NGS panel covering [23]. Furthermore, we proposed that macular progressive depigmentation with pigment clumping and atrophy is seen as a major complement in the proband, which was extremely apparent in the macula.…”

“…Eventually, all the detected variants of NGS in the present study were confirmed by PCR amplification and conventional Sanger-based sequencing (3500DX ABI genetic analyzer, Foster City, U.S.A.) in the proband and his family members [23–25]. In this regard, the locus-specific primers for variant verification and co-segregation analysis were designed using online Primer3 program.…”

Diagnostic value of a combination of next-generation sequencing, chorioretinal imaging and metabolic analysis: lessons from a consanguineous Chinese family with gyrate atrophy of the choroid and retina stemming from a novel OAT variant

“…Genomic DNA (gDNA) from peripheral leucocytes was extracted using a previously described phenol/chloroform method15 16 with DNA concentrations measured using a NanoDrop 2000 spectrophotometer 17. DNA was also extracted from samples of 100 individuals recruited as normal controls.…”

Choroideremia with Mutation in CHM Gene. Clinical Cases with Literature Review

“…В 2017 году L. Yang и соавт. разработали систему праймеров amplification refractory mutation system (ARMS) для быстрой и точной пренатальной диагностики хороидеремии методом полимеразной цепной реакции (ПЦР) [23].…”