ARMS-PCR Technique Research Articles

Single Nucleotide Polymorphism in the IFNG rs1861494 Gene among a Subset of Iraqi Pediatric Patients with Gastroenteritis Co-infected with Cryptosporidium and Adenovirus

Background: Research identifies enteric adenoviruses as the third most common cause of infantile gastroenteritis, while Cryptosporidium causes parasitic gastroenteritis. Many studies have revealed the role of IFN-γ in inflammation and autoimmune diseases. Objective: To investigate the IFNG rs1861494 gene polymorphism among Iraqi pediatric patients with gastroenteritis co-infected with cryptosporidium and adenovirus. Method: This case-control study enrolled 75 pediatric patients with severe gastroenteritis, whose ages ranged from 3–120 months and had a mean age of 30.64 months. The apparently healthy control (AHC) in this study included 25 pediatric individuals with a mean age of 27.64 months. We extracted DNA from stool specimens to further extract total genomic DNA, extract the human adenovirus (HADV) viral genome via PCR, and detect the IFNG rs1861494 polymorphism using the ARMS PCR technique. Results: The male gender percentage in patients and AHC groups was 54%, while the female counterpart was 46%. The positive PCR result for HADV7 was 28%, whereas the positive result for Cryptosporidium was 6%. In the studied groups, the GG genotype increased at a rate OR=2.67 as compared to the AG and AA genotypes. HADV7 and Cryptosporidium showed a strong correlation with the SNP IFNG rs1861494 results in gastroenteritis (r=0.968, p=0.007 and r=0.984, p=0.008). Conclusions: The identified enteric co-infection of Cryptosporidium and HADV-7, as well as the IFNG rs1861494 polymorphism, may shed light on possible pathogenic roles in gastroenteritis.

A Study on Associations of Long Noncoding RNA HOTAIR Polymorphisms With Genetic Susceptibility to Chronic Kidney Disease.

The importance of long noncoding RNAs (lncRNAs) in various biological processes has been increasingly recognized in recent years. This study investigated how gene polymorphism in HOX transcript antisense RNA (HOTAIR) lncRNA affects the predisposition to chronic kidney disease (CKD). This study comprised 150 patients with CKD and 150 healthy controls. A PCR-RFLP and ARMS-PCR techniques were used for genotyping the five target polymorphisms. According to our findings, rs4759314 confers strong protection against CKD in allelic, dominant, and codominant heterozygote genetic patterns. Furthermore, rs3816153 decreased CKD risk by 78% when TT versus GG, 55% when GG+GT versus TT, and 74% when GT versus TT+GG. In contrast, the CC+CT genotype [odds ratio (OR) = 1.66, 95% confidence intervals (CIs) = 1.05-2.63] and the T allele (OR = 1.50, 95% CI = 1.06-2.11) of rs12826786, as well as the TT genotype (OR = 2.52, 95% CI = 1.06-5.98) of rs3816153 markedly increased the risk of CKD in the Iranian population. Although no linkage disequilibrium was found between the studied variants, the Crs12826786Trs920778Grs1899663Grs4759314Grs3816153 haplotype was associated with a decreased risk of CKD by 86% (OR = 0.14, 95% CI = 0.03-0.66). The rs920778 was not correlated with CKD risk, whereas the HOTAIR rs4759314, rs12826786, rs1899663, and rs3816153 polymorphisms affected the risk of CKD in our population. It seems essential to conduct repeated studies across various ethnic groups to explore the link between HOTAIR variants and their impact on the disease outcome.

Study the Association between TP53 (rs12602273C>G) Gene Polymorphism with Susceptibility to the Breast Cancer in North West of Iran

Introduction: The TP53 gene on the short arm of chromosome 17p, which is known as a tumor suppressor gene, encodes the transcription factor p53, and this factor controls cell cycle progression. This study was done to assess the association of TP53 (rs12602273C>G) gene polymorphism with the risk and prognosis of breast cancer in women of northwest of Iran.
 Methods: In this case-control study, the association of TP53 (rs12602273C>G) gene polymorphism with breast cancer risk was studied in a population, including 100 patients and 100 healthy individuals was investigated by Tetra ARMS-PCR technique. The analysis of the resulting data was done using SPSS version 16 software and JavaStat online statistics package.
 Results: In the case group, the frequency of CC, CG, and GG genotypes were 81.52, 16.3, 2.17 percent, respectively and they were 79.34, 17.39, and 3.26 percent for the control group. Similarly, C and G allele frequency in case group was 89.67 and 10.32 percent and those in the control group was 88.04 and 11.95 percent, respectively. Statistical analyzes showed no association between genotypic (P>0.05) and allelic (P>0.05) polymorphism (rs12602273C>G) of TP53 gene with breast cancer. Also, the results showed that the studied polymorphism was not associated with the clinical characteristics of breast cancer patients in the Northwest of Iran (P>0.05).
 Conclusion: These research findings suggest that TP53 polymorphism of rs12602273C>G is not related to the risk of breast cancer. The association of the genotypic distribution of this SNP with the clinical characteristics of the patients was insignificant.

Novel and known minor alleles of CNTNAP2 gene variants are associated with comorbidity of intellectual disability and epilepsy phenotypes: a case-control association study reveals potential biomarkers.

Neurodevelopmental disorders are heterogeneous due to underlying multiple shared genetic pathways and risk factors. Intellectual disability, epilepsy and autism spectrum disorder phenotypes overlap which indicates the diverse effects of common genes. Recent studies suggested the probable contribution of CNTNAP2 gene polymorphisms to the comorbidity of these neurological conditions. This study was conducted to investigate the role of CNTNAP2 polymorphisms rs147815978 (G>T) and rs2710102 (A>G) as a risk factor for comorbidity of intellectual disability and epilepsy in a group of 345 individuals including 170 patients and 175 healthy controls recruited from various ethnic groups of Pakistani population. Our case-control study group was genotyped by tetra primer ARMS-PCR technique and results were analysed to know the effects of CNTNAP2 rs147815978 (G>T) and rs2710102 (A>G) polymorphisms in the group. The frequency of risk allele T (rs147815978) and risk allele G (rs2710102) for homozygous recessive genotypes (TT/GG) in our study group was 36.47% while odds ratios for risk allele T (rs147815978) was 5.45 (3.90-7.61: 95% CI, P = 0.000) and that for risk allele G (rs2710102) was 2.39 (1.76-3.24: 95% CI, P = 0.0001). Homozygous recessive genotypes (TT/GG) appeared only in cases and not in control group which indicated these as suspected risk genotypes and the significant association (p < 0.05%) of CNTNAP2 gene polymorphisms rs147815978 (G>T) and rs2710102 (A>G) with co-occurrence of intellectual disability and epilepsy phenotypes in our study group which is in HWE (χ2 = 174, P < 0.0001). Logistic regression analysis shows additive (p < 0.0001) and multiplicative (p < 0.001) models which confirms significant association of both the polymorphisms in our data, which are closely located on same haplotype (D' = -0.168). We propose that CNTNAP2 rs147815978 (G>T) and rs2710102 (A>G) polymorphisms are possible risk loci for overlapping neurodevelopmental disorders in Pakistani population. We propose the role of a previously reported common SNP rs2710102 (A>G) with a rarely reported novel SNP rs147815978 (G>T) for CNTNAP2 gene association with neurodevelopmental disorders in our data. Our study has expanded the knowledge of CNTNAP2 gene polymorphisms as probable biomarkers for susceptibility of co-occurrence of intellectual disability and epilepsy phenotypes in Pakistani population. We hope that our study will open new horizons of CNTNAP2 gene variants research to cure the neurological conditions in Pakistani population where consanguinity is a tradition and prevalence of neurodevelopmental disorders has increased from 1 to 2% during last 5 years.

SNP IFNG rs1861494 gene polymorphism in pediatric patients with gastroenteritis and its relation to co-infectious with Adenovirus and Cryptosporidium

Introduction and Aim: The most common cause of infantile gastroenteritis is enteric adenoviruses, while Cryptosporidiosis is caused by Cryptosporidium. The function of IFN-gamma (IFNG/IFN-y) in immunological responses, inflammation, and autoimmune disease is increasingly recognized. This study aimed to examine intestinal co-infections of Cryptosporidium and adenovirus in pediatrics and the role of IFNG rs1861494 polymorphism to gastroenteritis among Iraqi children. Materials and Methods: This case control study included 75 pediatric patients with severe gastroenteritis, aged 3-120 months, with a mean age of 30.64 months. The apparently healthy control (AHC) in this study included 25 pediatric individuals with a mean age of 27.64 months. Total genomic DNA extracted from stool specimens were subjected to PCR detection of human adenovirus virus. SNP IFNG rs1861494 polymorphism was detected using the ARMS-PCR technique. Results: The percentage of males to female ratio in this study was 54:46. The positive PCR result of HADV7 was 28% (14 of 50 cases) while the positive result of Cryptosporidium was 6% (6 of 75 cases).Genotyping studies showed the GG genotype increased at rate OR=2.67 as compared to AG and AA genotypes for the SNP IFNG rs1861494. The SNP IFNG rs1861494 results in gastroenteritis were highly correlated with HADV7 and Cryptosporidium (respectively: r = 0.968, P = 0.007 as well as r = 0.984, P = 0.008). Conclusions: Adenovirus diarrhea in Iraqi pediatric patients is associated with SNP rs1861494 polymorphism and could possibly be used as a biomarker in assessing the susceptibility of Iraqi children to the virus.

Impact of IL-6 and IL-1β Gene Variants on Non-small-cell Lung Cancer Risk in Egyptian Patients

Lung cancer is a serious health and life issue, with the fastest-growing incidence and fatality rates worldwide. It is now clear that inflammation is a key factor involved in all aspects of carcinogenesis, notably lung cancer development. Genetic changes, including polymorphisms in inflammatory genes, are supposed to be a significant cause of increased lung cancer risk. The main idea of this research was to disclose the linkage between both IL-6 rs1800795 and IL-1β rs16944 variants and susceptibility to non-small-cell lung cancer (NSCLC) in Egyptians. This case–control design was composed of 127 cases and 138 controls, which were genotyped using the ARMS-PCR technique. To examine the NSCLC susceptibility under various genetic models, the odds ratio (OR) and 95% confidence intervals (CIs) were determined by logistic regression. Rs1800795 of the IL-6 gene was linked to higher odds of NSCLC under the allele model (adjusted, OR 2.28; 95% CI 1.2–4.33; p = 0.011). In the genetic models, IL-6 rs1800795 elevated the odds of NSCLC, while IL-1β rs16944 decreased the odds of NSCLC. Stratification analysis showed that IL-6 rs1800795 greatly increased the NSCLC risk in females and adenocarcinoma subtypes, whereas IL-1β rs16944 largely decreased the NSCLC risk for males, patients aged < 55, and nonsmokers. Regarding clinical data, the IL-6 variant was remarkably correlated with tumor size. This work primarily established that IL-6 and IL-1β variants have a great impact on NSCLC development in the Egyptian population; thus, it may be a supportive guide for earlier NSCLC prevention.

Investigation of the Relationship between HIWI3 rs11703684 (C&gt;T) Polymorphism and Idiopathic Azoospermia/Oligozoospermia in the Kurdish Population of Kermanshah

Background &amp; Objective: Genetic factors play a major role in the development of male idiopathic infertility. Because of the multi-genetic base of this disease, not all genetic factors have been investigated. PIWI genes have been reported to be involved in the regulation of piRNAs in spermatogenesis. Our study assessed the association between HIWI3 rs11703684 (C&gt;T) gene polymorphism with the risk of male idiopathic azoospermia/oligozoospermia in the Kurdish population of Kermanshah.&#x0D; Materials &amp; Methods: In this case-control investigation, we included two hundred individuals consisting of 100 men with idiopathic azoospermia/oligozoospermia and 100 fertile men as the control group. To determine genotypes of HIWI3 C&gt;T polymorphism, we used the Tetra Arms-PCR technique and significant values were considered as p&lt;0.05.&#x0D; Results: Our findings did not show a statistically significant difference in the genotype frequency of the recessive model (P = 0.118; OR = 0.158; CI, 0.019–1.339), dominant (P = 0.169; OR = 0.625; CI, 0.341–1.144) and co-dominant (P = 0.527; OR = 0.778; CI, 0.417–1.450). In addition, the results described a negative difference in allelic frequency of HIWI3 (rs11703684 C&gt;T) in men with idiopathic azoospermia/oligozoospermia and control group (P = 0.288; OR = 0.749; CI, 0.463–1.212).&#x0D; Conclusion: The current study does not indicate the probability effect of HIWI3 rs11703684 (C&gt;T) gene polymorphism on the male idiopathic azoospermia/oligozoospermia in the Kurdish population of Kermanshah. The critical role of PIWI genes in spermatogenesis and as a candidate risk factor for male infertility remained unknown.

Allelic and Genotypic Analysis of LncRNA ANRIL rs4977574 A/G Mutations in Oral Squamous Cell Carcinoma Patients: Insights into Tumor Characteristics and Genotypic Correlations.

Long noncoding RNAs (lncRNA) ANRIL and its genetic polymorphisms are shown to be associated with the risk of several cancers. However, the single nucleotide polymorphisms (SNPs) of lncRNA ANRIL are not thoroughly assessed in oral squamous cell carcinoma (OSCC) which is the most prevalent cancer in the head and neck area. Thus, this study aimed to assess the association of SNP of lncRNA ANRIL rs4977574 in patients with OSCC. 106 blood samples from the patients with OSCC were obtained with a gender- and age-matched control group to evaluate the SNP of rs4977574 of lncRNA ANRIL. The DNA was extracted using the salt-out technique and DNA genotyping was undertaken using specific primer pairs in the tetra-primer ARMS-PCR technique. Eventually, the frequency of wild-type (A) and the mutated allele (G), as well as the genotypes were estimated between the groups of patients with OSCC and healthy individuals. The results of our study indicated no statistically significant difference in the frequency of rs4977574 A/G of lncRNA ANRIL among the patients with OSCC and healthy individuals (p > 0.05). Likewise, no significant difference was found in the genotypes' frequencies (p > 0.05). Nevertheless, the marked association of GG with smaller tumor size and the high level of differentiation of OSCC cells in the presence of AA or AG genotypes were interesting outcomes of this study (p < 0.05). Similarly, all the genotypes AA, AG, and GG were correlated with the site of the occurrence of OSCC. Furthermore, the association of the genotypes with the lymph node metastasis and the tumors stage was not found to be significant (p > 0.05). The results of our study indicate that rs4977574 A/G and its genotypes do not have any direct correlation with the presence of OSCC; however, its association with the smaller tumor size and the level of the cancer cells differentiation could imply its possible indirect role.

The Potential Impact of MYH9 (rs3752462) and ELMO1 (rs741301) Genetic Variants on the Risk of Nephrotic Syndrome Incidence.

The kidney lost a lot of protein in the urine when you have nephrotic syndrome (NS). Clinical manifestations mostly common in NS include massive proteinuria, hypoalbuminemia, hyperlipidemia, and edema. Idiopathic nephrotic syndrome is currently classified into steroid-dependent (SDNS) and steroid-resistant (SRNS) based on the initial response to corticosteroid therapy at presentation. Several reports examined the association of the MYH9 gene (rs3752462, C > T) variant and ELMO1 gene (rs741301 G > A) variant as risk factors for Nephrotic Syndrome. This study aimed to determine the potential effect of the MYH9 gene (rs3752462, C > T) and ELMO1 gene (rs741301) variant on the risk of (NS) among Egyptian Children. This study included two hundred participants involving 100 nephrotic syndrome (NS) cases and 100 healthy controls free from nephrotic syndrome (NS). The MYH9 gene (rs3752462, C > T) variant and ELMO1 gene (rs G > A741301) variant were analyzed by ARMS-PCR technique. Nephrotic syndrome cases include 74% SRNS and 26% SDNS. Higher frequencies of the heterozygous carrier (CT) and homozygous variant (TT) genotypes of the MYH9 (rs3752462, C > T) variant were observed in NS patients compared to the controls with p-value < 0.001. The frequencies of the MYH9 (rs3752462, C > T variant indicated a statistically significant elevated risk of NS under various genetic models, including allelic model (OR 2.85, p < 0.001), dominant (OR 3.97, p < 0.001) models, and the recessive model OR 5.94, p < 0.001). Higher frequencies of the heterozygous carrier (GA) and homozygous variant (AA) genotypes of ELMO1gene (rs G > A741301) variant were observed in NS patients compared to the controls with p-value < 0.001. The frequencies of the ELMO1 (rs G > A741301) variant indicated a statistically significant elevated risk of NS under various genetic models, including allelic model (OR 2.15, p < 0.001), dominant models (OR 2.8, p < 0.001), and the recessive model (OR 4.17, p = 0.001). Both MYH9 and ELMO1 gene variants are significantly different in NS in comparison with the control group (p < 0.001). The MYH9 gene (rs3752462, C > T) and ELMO1gene (rs G > A741301) variants were considered independent risk factors for NS among Egyptian Children.

Applying amplification refractory mutation system technique to detecting cell-free fetal DNA for single-gene disorders purpose.

Non-invasive prenatal diagnosis for single-gene disorders (NIPD) is still in development and deserves further study. The advent of next-generation sequencing technology significantly improved the detection of multiple mutations for non-invasive prenatal diagnosis for single-gene disorder purposes. However, bespoke amplicon-based NGS assays are costly. In this study, we developed a new strategy for non-invasive prenatal screening for single-gene disorders based on a capillary electrophoresis (CE) platform using an amplification refractory mutation system (ARMS)-PCR technique. Allele-specific primers for several disease-correlated mutations were designed, and subsequently, sensitivity and specificity assays were conducted. Assays on simulated two-person DNA mixtures showed that three primers targeting the mutant allele could detect minor DNA components in 1:500 mixtures. All primers showed positive results at 0.01ng of the template DNA. Cell-free fetal DNA was extracted from a pregnant woman's peripheral blood for the detection of paternally inherited mutations. Our results showed that one primer successfully amplified the mutant allele of fetal DNA in maternal plasma, which was confirmed by genotyping the genomic DNA extracted from amniotic fluid. This study suggested that the ARMS-PCR technique, a fast and cost-effective method, might be a promising method used to target de novo or paternally inherited pathogenic mutations in maternal plasma.

ZNF804A intronic variant rs1344706 and NGRN rs12807809 genetic polymorphisms and risk of schizophrenia in Bangladesh

BackgroundGenetic polymorphism exerts an undeniable impact on schizophrenia (SCZ) pathogenesis. Two polymorphisms, including rs1344806 of ZNF804A and rs12807809 of NRGN gene have been identified as susceptible candidates bearing the risk of SCZ. The target of this experimental study is to investigate the role of these polymorphisms with SCZ in native Bangladeshi people. MethodsFor this study, 220 schizophrenic patients and 204 non-schizophrenic controls were recruited to perform this investigation. For genotyping, tetra-primer ARMS-PCR technique was applied. The Agarose gel electrophoresis system visualized the final genomic products of the desired size. ResultsFrom the genotypic analysis of ZNF804A rs1344706 polymorphism, it was revealed that the minor allele frequency was noticeably increased in the patients than in the healthy population (42.27% vs. 29.17%). Five genetic models showed significantly increased risk of SCZ, including codominant model 1 (1.56-times), codominant model 2 (3.02-times), dominant model (1.86-times), recessive model (2.43-times), and allele model (1.78-times) (p < 0.05). The over-dominant model also had 1.18 times higher risk for SCZ, although the result could not meet the significance level (p > 0.05). Genotyping of rs12807809 unveiled the presence of a comparatively higher genotypic frequency of the minor allele among the patients than in controls (39.09% vs. 17.65%). Every genetic model under investigation showed significantly increased risk for SCZ– codominant model 1 (2.85-times), codominant model 2 (6.45-times), dominant model (3.44-times), recessive model (4.31-times), over-dominant model (2.10-times), and allele model (3.00-times) (p < 0.05). ConclusionThis experimental study concluded that rs1344806 of ZNF804A and rs12807809 of NRGN SNPs are significantly associated with SCZ in the Bangladeshi population.

Establishment of tetra-primer ARMS-PCR technique for detecting exo-E415G SNP in Plasmodium falciparum isolatesfrom Central Highlands of Vietnam

Drug resistance in Plasmodium falciparum parasites is associated with the single nucleotide polymorphism (SNP) exo-E415G, which is considered as a marker to reduce the susceptibility to artimisinin and leads to treatment failure. This study aims to establish a procedure based on tetra-primer ARMS PCR technique to detect the SNP exo-E415G in malaria patients who experienced treatment failure with artemisinin. DNA sample from P. falciparum 3D7 isolate was used to optimise the procedure based on tetra-primer ARMS PCR technique to detect exo-E415G SNP. The method was then applied to screen for exo-E415G SNP on 123 blood samples from patients who experienced treatment failure with artemisinin in 3 provinces (Gia Lai, Dak Nong, Dak Lak) from Central Highlands of Vietnam. A DNA fragment (749 bp) containing the exo-E415GSNP was amplified using primer pairs E415G-Fc and E415G-Rc. Specific primers for the wildtype alen (A alen) and the mutant alen (G alen) amplified fragments with the length of 316 and 474 bp, respectively. The genotyping results were consistent with Sanger sequencing. The tetra-primer ARMS PCR technique was used for genotyping exo-E415G SNP on 123 patients who failed with artemisinin treatment, the frequency of exo-E415G alen was observed at 78% (96/123). In conclusion, the simple tetra-primer ARMS PCR technique was established to detect the SNP Exo-E415G. The prevalence of the mutant alenExo-E415G was 78% in the Central Highlands of Vietnam.

Novel SNP Markers and Their Application in Low-Cyanide Cassava (Manihot esculenta crantz) Breeding Program

The major flaw of cassava is its indigenous cyanide. Traditional breeding program for diminishing cyanide in cassava take a long waiting time for harvesting cassava tubers to assess cyanide content. Genetic markers are useful for selection of good agricultural traits, which also applied in low-cyanide cassava breeding programs by detecting gene associate with cyanide content in cassava. Therefore, the objective of this study was to develop SNP markers to detect low cyanide trait in cassava. Genotype analysis was carried out using Genotyping by Sequencing (GBS) technology assembling with phenotype analysis. Then, the SNP markers associated with cyanide content were found. Afterward, the primers of SNP marker associated with cyanide content 2 positions were developed using tetra-primer ARMS-PCR techniques. S16_640082 and S16_735381 SNP markers on chromosome 16 showed potential ability of the selection of cassava varieties with cyanide content less than 250 mg HCN/kg fresh weight. The selection accuracy of S16_640082 and S16_735381 SNP markers was 73.33 and 76.64 %, respectively. These markers could be used in marker assisted selection (MAS) in cassava seedling. It substantially reduces cassava breeding programs cost and time. HIGHLIGHTS The major flaw of cassava is its indigenous cyanide. Up to our knowledge, there is rarely report on SNP marker related to cyanide content in Thai cassava population By performing genome wide association mapping (GWAS) to characterize the genome position related to HCN, 2 novel SNP markers associated with cyanide content were found The primers to detect the SNP markers were developed using tetra-primer ARMS-PCR techniques The markers showed ability of the selection of cassava varieties with cyanide content less than 250 mg HCN/kg fresh weight GRAPHICAL ABSTRACT

Genetic analysis of lncRNA H19 (rs217727) and MIAT (rs1894720) polymorphisms in patients with salivary gland tumors

Long noncoding RNAs (lncRNAs) represent a novel pharmacological application in cancer. Salivary gland tumors (SGTs) are essential to both maxillofacial and oral pathology. This study evaluated the H19 (rs217727) and Myocardial infarction-associated transcript (MIAT) (rs1894720) polymorphisms' connection to SGT susceptibility.We studied 104 Iranian patients with clinically definite SGTs and 114 healthy individuals. Samples collected from the participants were prepared, followed by DNA extraction. Following a tetra-primer ARMS-PCR technique employing specific primer pairs, DNA genotyping was carried out.The multivariate logistic regression analysis after adjusting the age revealed that the risk of SGTs was significantly reduced to a 0.17-fold in patients with TT genotype of rs217727 in the co-dominant model relative to CC genotype, while the risk of SGTs increased to 69-fold in patients with GT genotype of rs1894720 in the co-dominant model relative to GG genotype. In the dominant model, the risk of SGTs significantly were increased to a 41.8-fold in patients with GT + TT genotypes of rs1894720 relative to GG genotype. Also, in the recessive model, we found significant decrease in the risk of SGTs in patients with TT genotype of rs217727 compared to CT + CC and also in patients with TT genotype of rs1894720 relative to GT + GG genotype. In over-dominant model, the GT genotypes of rs1894720 significantly were associated to a 69-fold increase the risk of SGTs.Finally, the current study is the first to show that rs217727 and rs1894720 polymorphisms could be related to SGT risk in the Iranian population.

Study of single nucleotide polymorphism of aquaporin 2 in hypertension and CKD patients

The present study aims to examine the genetic variation of the Aquaporin-2 gene concerning hypertension and chronic kidney disease CKD. (120 ) blood samples from participants were obtained and divided into three groups: a first group representing hypertensive patients (hyper), a second group representing Hypertensive CKD patients (hyper with CKD), and a third group representing a control group. DNA was extracted from all blood samples and then converted to cDNA and the ARMS-PCR technique was used to investigate the single nucleotide polymorphism (SNP) from the AQP-2, genes. The result of this study investigated that SNP rs3759126 located in AQP-2 is a significant association with hypertension and CKD.

Polymorphism study of ATP1B1gene in hypertension with CKD patients

The present study aims to examine the genetic variation of the ATP1B1 gene concerning hypertension and chronic kidney disease CKD. (120 ) blood samples from participants were obtained and divided into three groups: a first group representing hypertensive patients (hyper), a second group representing Hypertensive CKD patients (hyper with CKD), and a third group representing a control group. DNA was extracted from all blood samples and then converted to cDNA and the ARMS-PCR technique was used to investigate the single nucleotide polymorphism (SNP) from the ATP1B1, genes. The result of this study investigated that SNP rs2901029 located in ATP1B1 is significant association with hypertension and CKD.

Evaluate IL-10 level and gene polymorphism in β-Thalassemia patients with related to microbial infection and disease outcome

Thalassemia is a genetic blood disorder inherited from the parent with unusual production of hemoglobin, infectious complications, immune abnormalities and iron overload with related to organ damage that consider a chief causes of morbidity and mortality. So this study aimed to evaluate the important causative agents and the role of IL-10 gene polymorphism in progressive disease. A Case-control study was performed to 46 Beta-thalassemia patients attended to inherited blood center in AL-Zahraa teaching hospital with group of 30 healthy individual as control. Blood sample was collected from all patients and control. IL-10 level were measured by enzyme-linked immunosorbent assay (ELISA). Singule nucleotide polymorphism detected by ARMS-PCR technique. The result shown that 46(46% ) patients with thalassemia , the male higher than female with frequency (65%), according to infection 32(70%) were TTI with HCV a most common pathogen 19(41 %).This study explain that serum level of IL-10 is significantly higher among patients with thalassemia (98±27.53)pg/ml compared to healthy control (10.67±1.49) pg\ml and was higher in HCV infected patients (121.56 ±32.60)pg/ml than non-infected with HCV record ( 73.61± 18.29) pg/ml.

Impact of IL-10-1082A\G promoter gene polymorphism and production in lung cancer patients

Lung cancer a deadliest malignant disease , Cytokines play a critical role in regulating host immune response toward cancer and determining the overall fate of tumorigenesis. So this study aimed to investigate the impact gene variation of IL-10 promotor region on production of IL-10 cytokine and their effect on lung cancer outcome. The aim of this study is the detection of IL-10-1082A\G polymorphism and IL-10 serum level with it correlation to progression of disease. A Case-control study was performed to 150 Lung cancer patients patients 54 females and 96 males and matched with age and gender attending to Middle Euphrates Cancer Center in AL-Najaf province with group of 60 healthy individual as control. Blood sample was collected from all patients and control. IL-10 level were measured by enzyme-linked immunosorbent assay (ELISA). Single nucleotide polymorphism detected by ARMS-PCR technique. The result shown that IL-10-1082 G/A significantly (P&lt;0.05) high frequencies of homozygous GG, and G allele carrier in lung cancer patients, in contrast homozygous AA and AG genotype and A allele appear at low frequency with patients. The data showed that genetic variants of the IL-10 affect the IL-10 production and associated with the progression of disease.

Lack of association between CYP27B1 gene polymorphism and type 2 diabetes mellitus in Iranian patients.

Type 2 diabetes mellitus (T2DM) is a global health problem with multiple etiological factors. Previous studies indicated that 1- alpha, 25-dihydroxyvitamin D3, a molecule that is produced by CYP27B1, could protect insulin-secreted cells from destruction by immune cells. The aim of the study was to investigate the CYP27B1 promoter gene polymorphism in T2DM. Two hundred subjects including 100 T2DM and 100 healthy individuals were recruited in the study. ARMS-PCR technique was used to identify rs10877012 genotypes in the 5' region of CYP27B1. The frequency of CC, CA, and AA genotype was 61/50, 31/39, and 8/11, respectively in T2DM patients compared to healthy subjects. There was no significant difference between both groups in regarding to genotype and allele distribution (P > 0.05). CA (RR = 1.535, 95% CI = 0.841- 2.802) and AA (1.677, 95% CI = 0.627-4.490) genotypes had no association with increased risk of T2DM. In addition, CA + AA versus CC showed no increased risk for T2DM (RR = 0.639, 95% CI = 0.365-1.121). We found no association between rs10877012 polymorphism and T2DM. There was no increased risk of this polymorphism in T2DM. Further studies with large groups are suggested in other populations to better understand the relation of CYP27B1 gene variation, especially its ethnicity-dependent relation with T2DM.

Allele Frequency of APAF1 Mutation in Holstein Cattle in Brazil.

APAF1 is an autosomal recessive inherited mutation, associated with Holstein haplotype 1 (HH1) and characterized by a substitution of cytosine for a thymine (c.1741C>T) in chromosome 5. The mutation causes fetal and embryonic loss, between 60 and 200 days of gestation, and reduced conception rate. The ARMS-PCR is considered a simple and low-cost method to determine single nucleotide polymorphism (SNP) with no need for genetic sequencing of the animal genome. This study aimed to verify the allelic frequency of APAF1 mutation in Brazilian Holstein cattle. A total of 248 Holstein DNA samples (210 cows and 38 bulls) were analyzed, and synthetic genes were manufactured to validate the primers developed by the authors. All animals assessed in this study were classified as wild-type for APAF1 mutation. The primers and protocol developed for the ARMS-PCR technique work with 100% specificity and efficiency since the amplicon formations are as expected according to the genotypes. In conclusion, the mutation responsible for APAF1 was not detected in the Brazilian Holstein cattle population assessed in this prevalence study, although it is not possible to affirm that APAF1 does not occur in Brazilian Holstein animals. The tetra-primer ARMS-PCR protocol for APAF1 mutation that has been validated here may be a relatively simple and economical method to determine the animals' genotype.