A SNaPshot assay for detection of 45 mutations in the SCN5A gene in the Chinese Han Population

Abstract

Sudden cardiac death (SCD) occurs frequently in forensic practice and results in no visible pathological changes that can be detected in an autopsy. In recent years, the genetic background has been emphasized when examining SCD cases. The aim of this study is to establish a feasible system to detect SCD-related genes for forensic DNA laboratories. Forty-five reported SCD-associated SNPs from sodium voltage-gated channel alpha subunit 5 (SCN5A) were considered in our experiment. We established a SNaPshot assay for the typing of 45 SNPs using multiplex PCR and the minisequencing technique. Two multiplex PCRs were performed and optimized to cover 14 and 16 DNA fragments. The SCD victims came from the Chinese Han population residing in Shanxi and Chongqing provinces and were examined and compared with a non-SCD group and with normal healthy individuals. A missense mutation at rs1805124 (H558R) was detected in the Chinese Han population in this study. A SNaPshot assay can be performed in any forensic DNA laboratory and would be capable of meeting the increasing demand for SCD detection. This method would also be beneficial for screening at-risk in family members of SCD victims.

Predicting human age by detecting DNA methylation status in hair

“…The PCR conditions were 95°C for 5 min followed by 35 cycles at 95°C for 30 s, 54°C for 30 s, and 72°C for 30 s. The final extension was 72°C for 10 min. Then, the PCR products were purified and extended by using the same conditions described by Wang et al [37,38].…”

Screening of PAH Common Mutations in Chinese Phenylketonuria Patients Using iPLEX MALDI-TOF MS

“…This indicates the need for new ways to detect common PAH mutations. Recently, several high-throughput technologies, such as Denaturing high performance liquid chromatography (DHPLC), high resolution melting (HRM), SNaPshot, and amplification-refractory mutation system-polymerase chain reaction (ARMS-PCR) analysis, have been developed. One such promising technology called iPLEX MALDI-TOF MS, which is based on matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry and provided by Sequenom Inc. (San Diego, CA), is also known as MassArray technology.…”

“…This indicates the need for new ways to detect common PAH mutations. Recently, several high-throughput technologies, such as Denaturing high performance liquid chromatog- raphy (DHPLC), 6 high resolution melting (HRM), 7 SNaPshot, 8 and amplification-refractory mutation system-polymerase chain reaction (ARMS-PCR) 9 S1. The remaining six mutations (c.208_210del, c.694C > T, c.739G > C, c.740G > T, c.482T > C, and c.1252A > C) were excluded from the assay because of their low frequencies and interference of the primers used to detect the mutations with other primers in the multiplex reaction.…”