Evaluation of a modified salt-out method for DNA extraction from whole blood lymphocytes: A simple and economical method for gene polymorphism

Shokrzadeh, M; Mohammadpour, Abbas

doi:10.18502/pbr.v4i2.218

Cited by 10 publications

(7 citation statements)

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“…Genomic DNA was extracted by the manual salting-out method from 2 ml of peripheral blood samples containing EDTA (1.8 mg EDTA per milliliter of blood) [12]. The quality and quantity of extracted DNA were measured using Epoch™ microplate spectrophotometer (Biotek Instrument, USA).…”

Section: Dna Extraction and Genotype Analysismentioning

confidence: 99%

Association Between Retinoid X Receptor Gene Variants and Dyslipidemia Risk in an Iranian Population

Vafaeie¹,

Toba²,

Khosravi³

et al. 2021

Cureus

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BackgroundDyslipidemia is a complex trait that is influenced by various genetic and environmental factors. While the exact cause of dyslipidemia is still unknown, some studies have shown that genetic factors such as single nucleotide polymorphisms (SNPs) have been primarily associated with dyslipidemia. Based on the available data, it appears that retinoid X receptor (RXR) genes are jointly or separately associated with lipid homeostasis and that SNPs may affect RXR gene functions in lipid metabolism. MethodsTo study the possible role of the RXR genes in genetic susceptibility of dyslipidemia, three selected polymorphisms, rs3132294 located in RXRA (RXR-alpha) gene and rs2651860 and rs1128977 located in RXRG (RXR-gamma) gene, were investigated in 391 individuals with the use of tetra-primer amplification refractory mutation system polymerase chain reaction (T-ARMS PCR) method. ResultsFor the rs3132294 SNP, the genotype frequencies in the case group were GG 58.5%, GA 33.2%, and AA 8.3%, and in the control group, they were GG 51.8%, GA 36.3%, and AA 11.9%. The genotype distribution of rs2651860 SNP in the case group were TT 43.2%, TG 52.1%, and GG 4.7%, and in the control group, they were TT 50.8%, TG 46.2%, and GG 3%. Genotype frequencies for the rs1128977 SNP in the case group were CC 34.7%, CT 47.6% and TT 17.7%, compared with CC 37.8%, CT 44.3%, and TT 17.9% in the control group. When the clinical characteristics of the case and control groups were stratified by allele carrier status for each SNP, the rs1128977 SNP was associated with increased levels of HDL-cholesterol, body mass index, waist circumference, and diastolic blood pressure (P< 0.05). In contrast, the alleles of the rs2651860 and rs3132294 SNP were not associated with an increased prevalence of dyslipidemia or clinical characteristics in the case group compared to the control group. ConclusionThe present study suggests that rs1128977 SNP in the RXRG gene may affect the clinical characteristics in cases. However, further genetics association studies on large samples are required to validate our findings.

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Section: Dna Extraction and Genotype Analysismentioning

confidence: 99%

Association Between Retinoid X Receptor Gene Variants and Dyslipidemia Risk in an Iranian Population

Vafaeie¹,

Toba²,

Khosravi³

et al. 2021

Cureus

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“…MTHFR C677T polymorphism analysis MTHFR C677T polymorphism was detected in DNA extracted from whole blood by the use of real-time PCR (TaqMan® assay). The DNA was extracted from 300 ul of blood using the standard salting out method [17].…”

Section: Study Population and Samplesmentioning

confidence: 99%

Association of MTHFR C677T variant genotype with serum folate and Vit B12 in Iranian patients with colorectal cancer or adenomatous polyps

Ghorbani

Azghandi

Khayyami

et al. 2021

Preprint

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Background: The incidence of colorectal cancer (CRC) has increased during recent years in Iran and other developing countries. Clinical studies suggest that essential folate dietary intake and moderate deficiency of methylenetetrahydrofolate reductase (MTHFR) may protect and reduce the risk of CRC.The aim of the present study was to investigate the clinical significance of C677T polymorphism within MTHFR gene and its correlation with the serum folate and Vit B12 in the Iranian population suffering from CRC.Methods: Blood samples were taken from 1017 Iranian individuals (517 cases and 500 controls) who were referred for colonoscopy. TaqMan probe assay was performed for C677T MTHFR polymorphism. Sera were fractionated from the blood samples of 43 patients and controls and folate and Vit B12 concentrations were measured by a monobind kit. The correlation of MTHFR polymorphisms and folate/vitamin-B12 with CRC risk were analyzed.Results: In the current study, we found the frequency of three different genotypes of MTHFR polymorphism in the Iranian population i.e., CC, CT and TT, to be 51.31, 26.73, 21.96 and 61, 32.2, 6.8 in case and control groups, respectively. The homozygote genotype of MTHFR rs1801133 polymorphism is associated with increased risk of CRC by 3.68, 1.42 and 3.74-fold in codominant, dominant and recessive models respectively (p value< 0.01). Our study revealed that there was no significant difference between the amount of folate and Vit B12 in case and control groups (p value> 0.05).Conclusions: This study revealed that there was no significant difference between the amount of folate and Vit B12 in the case and control groups. Furthermore, our results demonstrated a higher risk association for 677TT and 677TT+C677T genotypes of MTHFR compared with 677CC carriers among CRC patients.

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“…Total genomic DNA was extracted from the leukocytes using DNA isolation kit (Denazist Asia Co., Mashhad, Iran). The concentration and purity of the extracted DNA were measured using a Nanodrop spectrophotometer (Biochrom WPA Biowave II + UV / visible spectrophotometer, Cambridge, UK) 20 . The Arg389Gly and Ser49Gly polymorphisms in the β 1 AR were confirmed by polymerase chain reaction-restriction fragment length polymorphism (RFLP-PCR) methods.…”

Section: Clinical Data Collectionmentioning

confidence: 99%

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2021

IJPSR

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Postoperative atrial fibrillation (POAF) is a frequent rhythm disturbance after cardiac surgery and is associated with poor cardiovascular outcomes. Beta-blocker therapy is an effective strategy to prevent or decrease the high incidence of POAF and β 1 -adrenergic receptor (β 1 AR) polymorphisms that affect the cardiac response to β 1 AR antagonists. The study objective is to analyze the association between genetic variations in the β 1 AR gene and the risk of POAF and the prophylactic effect of beta-blockers. Totally, 198 patients undergoing first-time cardiac surgery were examined. The patients after operation were grouped as either with POAF or without POAF according to their continuously electrocardiogram monitoring within ICU and hospital admission period. Subsequently, genotype frequencies of the β 1 AR gene were determined by polymerase chain reaction-restriction fragment length polymorphism. The results showed that 63 patients (31.8%) of the total study population developed POAF. After the application of a multivariate logistic regression model, the incidence of POAF was associated with Arg389Gly genotype (OR=2.48; 95% CI, 1.06-5.76; p=0.036), Arg389Arg genotype (OR=3.38; 95% CI, 1.28-9.33; p=0.018) and Arg389Ser49 haplotype (OR=1.73; 95% CI, 1.07-2.80; p=0.027) in the whole population. In addition, subgroup analysis showed an association between Arg389Gly (OR=3.09; 95% CI, 1.12-8.52; p=0.03) and Arg389Arg (OR=4.72; 95% CI, 1.07-15.43; p=0.01) genotypes as well as Arg389Ser49 haplotype (OR=2.05; 95% CI, 1.16-3.62; p=0.013) with increased risk for POAF in patients using beta-blocker prophylaxis. These findings indicated that POAF might be related to the Arg389-β 1 AR variant in the studied Iranian population, and this association appears to be more prominent among patients with betablocker prophylaxis.

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Evaluation of a modified salt-out method for DNA extraction from whole blood lymphocytes: A simple and economical method for gene polymorphism

Cited by 10 publications

References 0 publications

Association Between Retinoid X Receptor Gene Variants and Dyslipidemia Risk in an Iranian Population

Association Between Retinoid X Receptor Gene Variants and Dyslipidemia Risk in an Iranian Population

Association of MTHFR C677T variant genotype with serum folate and Vit B12 in Iranian patients with colorectal cancer or adenomatous polyps

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