Schizophrenia is a severe neuropsychiatric disorder with symptoms such as hallucination, delusion and mental disorder. It is a complex disorder, in which genetic components play a crucial role in its pathogenesis. Among candidate genes for schizophrenia, Neuregulin 1 (NRG1) gene is the most important gene, association of which with the illness has been confirmed in several studies. Single nuclotide polymorphisms (SNPs) located 5´ upstream of NRG1 have shown significant association with schizophrenia in several populations. Here, we describe a designed simple Multiplex TETRA-PRIMER AMPLIFICATION REFRACTORY MUTATION SYSTEM - polymerase chain reaction (PCR) for genotyping single SNP (SNP8NRG221533) in the human NRG1 gene. No restriction site was found for distinguishing T and C alleles of this SNP. The developed METHOD proved to be simple, rapid and cost effective. This technique was used to compare SNP8NRG221533 in 95 schizophernics and 95 healthy controls. Our data demonstrate that there is a significant difference between allelic and genotypic frequencies of the two groups. These preliminary results confirm the association of the NRG1 gene with schizophrenia in an Iranian population.

Background: Dyslipidemia refers to an abnormal level of plasma lipids, mainly high levels of low-density lipoproteins and triglycerides, and low levels of high-density lipoproteins. This lipid abnormality is demonstrated to be strongly linked to the development of cardiovascular diseases. Genome-wide association studies have shown several single nucleotide polymorphisms (SNPs) linked with dyslipidemia. Materials and METHODs: In this investigation, we have optimized a TETRA-PRIMER AMPLIFICATION REFRACTORY MUTATION SYSTEM ((T-ARMS)) polymorphism chain reaction (PCR) protocol for four SNPs, i. e., CELSR2 (rs629301), APOB (rs1367117), APOE (rs429358), and APOE (rs7412) associated with dyslipidemia. Results: (T-ARMS) PCR is a multiplex, sensitive, and low-technique to genotype the SNPs performed in a single tube and reaction. Several factors, such as DNA purity, annealing temperature, PCR reagent equilibrium, and concentration of magnesium chloride in the reaction, influence the assay. However, the (T-ARMS) PCR assay has been demonstrated to be unsuitable for variants rich in guanine and cytosine. Conclusion: (T-ARMS) PCR is the METHOD of choice for association studies of variants with genetic diseases and provides a fast, accurate, reliable, and low-cost way to study SNPs.

Background: b -thalassemia, a monogenic autosomal recessive disorder, is prevalent in Middle East, particularly in Iran. In Iran, near to 20 MUTATIONs in the b-globin gene are introduced as common MUTATIONs with varying incidence frequencies in each city. Therefore, detection and screening for couples at high risk can help to solve the problems of this disease. In this study, optimized genotyping of two common MUTATIONs in Isfahan Province, IVSII-I (G-A) and FSC-8/9 insG, was performed using the (T-ARMS) METHOD.METHODs: In this case-control study, 10 healthy individuals and 30 patients affected by β-thalassemia major with a mean 24.76+4.5 years were selected from Omid Hospital in Isfahan Province. After designing TETRA PRIMERs for two prevalent MUTATIONs IVSII-I (G-A) and FSC-8/9 insG, samples were genotyped using TETRA-PRIMERs ARMS PCR technique.Results: We have developed a sensitive single tube TETRA-PRIMERs PCR assay to detect both IVSII-1 (G-A) and FS8-9 insG MUTATIONs. Moreover, we have distinguished homozygous and heterozygous forms of these MUTATIONs successfully. The frequency of IVSII-1 (G-A) MUTATION from 30 patients in Isfahan was 86.6% (33.3% heterozygote, and 53.3% mutant homozygote) and for FS8-9 insG MUTATION was 16.6% (13.3% heterozygote, and 3.3% mutant homozygote).Conclusion: TETRA-PRIMERs ARMS PCR could be a reliable, accurate and simple technique for genotyping SNP and different MUTATIONs. So far, no study was done on optimization METHODs for genotyping MUTATIONs in b-thalassemia by (T-ARMS). Here, we successfully adjusted and enhanced this METHOD for recognizing two common MUTATIONs (FSC-8/9 insG and IVSII-I (G-A)) of b-thalassemia in Isfahan population.

Background: Crigler-Najjar syndrome is a rare, autosomal recessive disorder characterized by unconjugated, non-hemolytic hyperbilirubinemia. The disease is caused by MUTATIONs in the UGT1A1 gene, which result in the decrease or lack of the UGT1A1 enzyme activity and thus, lack of bilirubin conjugation with glucuronic acid. Molecular diagnosis of the syndrome has been essentially based on direct MUTATION analysis. However, due to a large number of MUTATIONs associated with the disease, direct MUTATION analysis is expensive and time consuming. Alternatively, indirect analysis of MUTATIONs using linkage analysis by means of polymorphic markers is proposed. Several polymorphic markers associated with theUGT1A1 gene have been studied.METHODs: Using bioinformatic analysis of these markers, the single nucleotide polymorphism (SNP) rs4148326 with the C/T sequence, located at 5' region of the gene was selected. Analysis of the marker was performed by genotyping of 186 unrelated healthy individuals in the Isfahanian population, Iran, using TETRA-PRIMER AMPLIFICATION REFRACTORY MUTATION SYSTEM polymerase chain reaction (TETRA-PRIMER ARMS PCR) technique. Statistical analysis of the results was performed using the GENEPOP software.Findings: The heterozygosity of rs4148326 the marker was 62.9% and the allele frequency for T and C allele was 66.94% and 33.06%, respectively. Analysis of deviations from Hardy-Weinberg equilibrium demonstrated that the Isfahanian population was in equilibrium (P<0.001) for rs4148326 locus.Conclusion: The data suggested that rs4148326 could be introduced as an informative marker for molecular diagnosis of Crigler-Najjar syndrome in a representative sample of the Iranian population.

Background and Objective: Prostate cancer is among the five common cancers in males. It is second cancer in terms of the age-standardized rate (ASR) (ASR=16. 6) in Iran. The rs2735839 G/A, an intergenic polymorphism is located on chromosome 19q13. 33 at 600 base pairs of the KLK3 gene untranslatable region. This gene which codes prostate-specific antigen (PSA) is used in the screening and diagnosis of prostate cancer. The purpose of this study was to evaluate the association between this polymorphism and prostate adenocarcinoma with PSA. Materials and METHODs: This case-control study included 103 and 100 patients with prostate adenocarcinoma and benign prostatic hyperplasia (BPH) as case and control groups, respectively. TETRA-PRIMER AMPLIFICATION REFRACTORY MUTATION SYSTEM-polymerase chain reaction was used to determine the genotype of each participant regarding rs2735839 polymorphism. Results: There was a significant difference between the adenocarcinoma prostate and BPH groups regarding genotype frequency AG+AA (OR [95% CI]=4. 991 [2. 475-10. 065], P=0. 00). According to the results of statistical analysis, a significant difference was observed between the adenocarcinoma and BPH groups in terms of allele frequency (OR [95% CI]=3. 927 [2. 085-7. 397], P=0. 00). Moreover, There was a significant difference between rs2735839 and PSA regarding the genotype frequency polymorphism (P=0. 011). Conclusion: The results indicate that rs2735839 is associated with an increased risk of prostate cancer in Iranian population. It is worth noting that a significant difference was found between the distribution of allele A and that of allele G with PSA levels of >10.

Background & Aims: Tyrosinase is the most important enzyme in the production of pigments of the skin, eyes, and hair follicles. The enzyme is encoded by tyrosinase gene (TYR) or oculocutaneous albinism type 1A (OCA1A). MUTATIONs in TYR gene result in pigmentation disorders such as albinism in humans. In view of the large number of MUTATIONs reported in this gene, the aim of this study was to identify and introduce polymorphic markers located in the TYR gene region in the Iranian population.METHODs: In the present study, using bioinformatics investigations for single nucleotide polymorphisms (SNP) markers in TYR gene region, rs1799989 marker was selected and genotyped through TETRA PRIMER AMPLIFICATION REFRACTORY MUTATION SYSTEM-polymerase chain reaction (ARMS-PCR) METHOD. The allele frequency and heterozygosity degree of this marker was analyzed in the population of Isfahan, Iran.Results: The heterozigosity of the rs1799989 marker (75.9%) in the population of Isfahan was high. The frequency of allele C and allele A was 0.576 and 0.420, respectively. Comparison of this finding to those of other populations showed that the most similarity existed with African populations.Conclusion: In view of the high degree of heterozygosity, rs1799989 marker can be introduced as an informative marker for linkage analysis and identification of TYR gene MUTATIONs carriers in the Iranian population.

Background: Genetic polymorphisms in DNA-repair genes may increase the risk of developing cancer due to reducing the DNA-repair capacity. XRCC1 is one of the important genes in DNA repair. This study was designed to examine the polymorphisms associated with XRCC1 Arg399Gln and to investigate its role as susceptibility marker for non-small cell lung cancer (NSCLC) in population of Fars province, Iran.METHODs: In this case-control study, extracted DNA from 100 healthy controls and 100 patient of lung cancer were used to examine the role of XCRR1 Arg399Gln polymorphism in context of non-small cell lung cancer in population of Fars province in Iran. AMPLIFICATION-REFRACTORY MUTATION SYSTEM-polymerase chain reaction (ARMS-PCR) technique was used for determining of individuals’ genotyping.Findings: Our data showed a strong association between Gln/Gln (A/A) genotype and risk of developing non-small cell lung cancer in men. Moreover, men with at least one A allele (AA+AG) showed reduced risk of developing non-small cell lung cancer. No such an associations were found in subgroups of women or when samples were divided based on their ages.Conclusion: According to our results, although there was no significant association between XRCC1 Age399Gln polymorphism and developing of lung cancer in population, men with Gln/Gln genotype were in high risk of developing non-small cell lung cancer in Fars province, Iran. Therefore, Gln/Gln polymorphism could be used as a biomarker for screening men at high risk of lung cancer.

Background-Beta-thalassemia is the most common hereditary disorder in Iran and during the past 10 years, AMPLIFICATION REFRACTORY MUTATION SYSTEM (ARMS) and restriction fragment length polymorphism (RFLP) were the sole molecular technique used for diagnosis of the disease. Although many beta-globin gene MUTATIONs exist in the Iranian multiethnic population, these techniques seem labor-intensive, time-consuming and expensive. This has urged us to use new techniques such as reverse hybridization and direct sequencing this issue. METHODs-In this study, reverse hybridization was applied in parallel with ARMS to screen for the 10 most common beta-thalassemia MUTATIONs and hemoglobin S in 82 patients clinically diagnosed as beta-thalassemia minor and major. Results-From the 82 cases detectable by both METHODs, 80 had similar results. Compared to ARMS, reverse hybridization appeared to be more reliable, cost-effective, fast and applicable. Conclusion-Considering the vast spectrum of beta-thalassemia MUTATIONs in Iran, a fast and reliable technique such as reverse hybridization represents vital advantages in comparison with the traditional diagnostic METHODs. In fact, it is recommended as the technique of choice that can be employed by the National Thalassemia Project for the detection and prenatal diagnosis of beta-thalassemia in Iran.