Tetra-ARMS-PCR assay development for genotyping of <i>AGT</i> rs699 T/C polymorphism, its comparison with PCR-RFLP and application in a case-control association study of cardiovascular disease patients

Hussain, Misbah; Khan, Haq Nawaz; Abbas, Shahid; Ali, Ansar; Aslam, Muhammad Naeem; Awan, Fazli Rabbi

doi:10.1080/15257770.2023.2181972

Cited by 4 publications

(1 citation statement)

References 39 publications

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“…Tetra Amplification Refractory Mutation System Polymerase Chain Reaction (T-ARMS PCR) was used for the identification of the reported SNP in the collected samples. Using PCR amplification and subsequent gel electrophoresis is an allele specific, fast, reliable, accurate, and inexpensive way to identify an SNP [12,13]. Applying this technique, four primers were used under a single PCR reaction, namely forward outer primer, reverse outer primer, forward inner primer, and reverse inner primer.…”

Section: Methodsmentioning

confidence: 99%

Development of T-ARMS-PCR to Detect MYBPC3 Gene Variation in Hypertrophic Cardiomyopathy (HCM) Patients

Sadia,

Ahmed Khan,

Hussain

et al. 2023

CTO

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Hypertrophic cardiomyopathy (HCM) is a common and complex, genetically inherited, cardiovascular disorder. It is typically inherited in an autosomal dominant manner with variable penetrance and mutable expression. Mutations in MYBPC3 gene is one of the genetic causes of HCM. Only 0.2% of general population suffers from HCM. The MYBPC3 gene provides instructions for making cardiac myosin binding protein C, which is imperative for the maintenance and regulation of normal cardiac functions. This study aims to explore the reported SNP rs1052373 from exon 30 of MYBPC3 gene in the population of Punjab, Pakistan. The reported SNP rs1052373 was analysed using Tetra Amplification Refractory Mutation System Polymerase Chain Reaction (T-ARMS-PCR) to find the allelic frequency in the selected population. T-ARMS-PCR is a cost effective, flexible, rapid, and accurate tool for genotyping. The specific sequences of MYBPC3 gene from exons 30 and 31 and introns 29, 30, and 31 were retrieved from NCBI (https://www.ncbi.nlm.nih.gov/). A tetra primer designing tool known as Primer 1 (http://primer1.soton.ac.uk/primer1.html) was used to design the primers for the targeted region of MYBPC3 gene. In this study, the genotyping of previously reported SNP rs1052373 showed variation in the disease group, giving CC, CT, and TT genotypes with the frequency of 0.04. The genotyping analysis of rs1052373 showed that the allelic frequency of homozygous condition T/T was 0.02 and the allelic frequency of heterozygous condition C/T was 0.02 in disease group as compared to the control group. In the latter, the homozygous T/T and heterozygous C/T genotypes were not observed in any individuals. All the individuals in control group carried homozygous C/C genotype. While, the frequency of homozygous C/C genotype was 0.96 in disease group. The findings of this study would help to find novel molecular markers for HCM diagnosis.

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Section: Methodsmentioning

confidence: 99%

Development of T-ARMS-PCR to Detect MYBPC3 Gene Variation in Hypertrophic Cardiomyopathy (HCM) Patients

Sadia,

Ahmed Khan,

Hussain

et al. 2023

CTO

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Genetic screening for pathogenic variants in type 2 diabetes of the Arab Gulf population: A systematic review and meta-analysis

Musafer,

Rava,

Al-Shuhaib

2023

Int J Diabetes Dev Ctries

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Development of a tetra-primer ARMS–PCR for identification of sika and red deer and their hybrids

Ke-xin,

Xiang,

Qing-qing

et al. 2023

ANAL. SCI.

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Accurate identification of deer-derived components is significant in food and drug authenticity. Over the years, several methods have been developed to authenticate these products; however, identifying whether female deer products are hybrids is challenging. In this study, the zinc finger protein X-linked (ZFX) gene sequences of sika deer (Cervus nippon), red deer (Cervus elaphus) and their hybrid offspring were amplified and sequenced, the X221 and X428 species-specific single nucleotide polymorphisms (SNP) loci were verified, and a tetra-primer amplification refractory mutation system (T-ARMS–PCR) assay was developed to identify the parent-of-origin of female sika deer, red deer, and their hybrid deer. The T-ARMS–PCR developed based on the X221 locus could identify sika deer, red deer, and their hybrid offspring according to the presence or absence of PCR product sizes of 486 bp, 352 bp, and 179 bp, respectively, just as X428 locus could identify sika deer, red deer, and their hybrid offspring according to the presence or absence of PCR product sizes of 549 bp, 213 bp, and 383 bp, respectively. Forty products labeled deer-derived ingredients randomly purchased were tested using this assay, and the results showed that the identification results based on the two SNP loci were utterly consistent with the actual sources. In addition, this method was found to be accurate, simple, convenient, and with high specificity, thus providing an essential technical reference for deer product species identification. It is also an important supplement to the identification methods of the original ingredients of existing deer products. Graphical abstract

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Tetra-ARMS-PCR assay development for genotyping of AGT rs699 T/C polymorphism, its comparison with PCR-RFLP and application in a case-control association study of cardiovascular disease patients

Cited by 4 publications

References 39 publications

Development of T-ARMS-PCR to Detect MYBPC3 Gene Variation in Hypertrophic Cardiomyopathy (HCM) Patients

Development of T-ARMS-PCR to Detect MYBPC3 Gene Variation in Hypertrophic Cardiomyopathy (HCM) Patients

Genetic screening for pathogenic variants in type 2 diabetes of the Arab Gulf population: A systematic review and meta-analysis

Development of a tetra-primer ARMS–PCR for identification of sika and red deer and their hybrids

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