Hepatitis C virus (HCV) is a major global health problem and one of the most common causes of chronic liver diseases. The current study was carried out to investigate the ability of High Resolution Melting (HRM) Method in HCV Genotyping. 40 HCV-Positive sera with unknown and 12 sera with known HCV genotypes were collected from different clinical laboratories in Tehran as well as Rasht in north of Iran. The RT-PCR was performed on RNA extracted from each sample. The first PCR amplification product was used as template for second run in a Nested-PCR followed by HRM Method. Unknown genotypes were determined with comparing of their normalized and difference graphs with those obtained by known genotypes as standard. All results were confirmed by direct sequencing. In a comparison between results obtained by HRM Method and those of direct sequencing, only 2 out of 40 samples with unclear HCV genotypes had been incorrectly genotyped by HRM Method. The results obtained by this study show that HRM can be introduced as a simple, fast, and reliable Method for HCV Genotyping in clinical laboratories.

In recent decades, different Methods have been introduced for the Genotyping of Sin-gle Nucleotide Polymorphisms (SNPs) and mutations in nucleic acid sequences. These Methods have several applications ranging from agriculture to medicine. The Loop-mediated isothermal amplification (LAMP) Method was first introduced by Notomi et al. Since then, different Methods derived from LAMP have been extensively applied in detecting pathogens. The LAMP Method is an isothermal technique that amplifies the target DNA segment using four different primers that have been uniquely designed for recognizing six distinct zones on the objective gene; the process of reaction contin-ues at a constant temperature via a strand displacement reaction. Amplifying and detecting the targeted zone can be accomplished in one stage. Although the LAMP Method is mostly used for pathogen detection, several studies have used this Method for Genotyping. The present article reviewed various studies that used the LAMP Method for SNP detection. The outcomes indicated that the LAMP technique could be a reliable and alternative technique for Genotyping. Further studies are recom-mended to use this approach for Genotyping.

Background: Mycobacterium tuberculosis Genotyping is an essential step for understanding the epidemiology of tuberculosis. Mycobacterial-interspersed-repetitive-units (MIRU)-variable-number-of-tandem-repeats (VNTR) typing is an important Method for this purpose. Objectives: The study aimed to determine the reproducibility of 15-loci MIRU-VNTR Method. Methods: DNA extraction and 15-loci MIRU-VNTR were carried out for Genotyping of 60 M. tuberculosis isolates collected from different clinical samples of 27 M. tuberculosis patients, each with two to four clinical isolates of M. tuberculosis. The similarity of M. tuberculosis isolates of the same patient was investigated by MIRU-VNTRplus. Results: The patterns of MIRU-VNTR were identical in 43 M. tuberculosis isolates collected from 20 tuberculosis patients, giving the repeatability of 71. 6% for the test. In 12 isolates of M. tuberculosis belonging to five patients, the variable-number tandem repeat (VNTR) in only one locus was different. In three M. tuberculosis isolates of one patient, two different genotypes were identified. Conclusions: This study confirms the high reproducibility of 15-loci MIRU-VNTR, except for limited cases.

Background: Genetic polymorphisms of drug metabolisms by cytochrome P450 (P450s) could affect drug response, attracting particular interest in the pharmacogenetics.Due to the importance of CYP2C19* 17 allele and its capability of super- fast metabolism and also lack of information about distribution of the alleles in Iranian population, this research aimed to use High Resolution Melting (HRM) Method compared to PCR-RFLP for Genotyping healthy Iranian population.Methods: Blood samples were collected from 100 healthy Iranian volunteers. DNA was extracted by salting out Method. Real-time PCR was used for amplification of the CYP2C19 gene and the alleles were identified by HRM. Sequencing was used to confirm the amplified DNA fragments and data were analyzed using SPSS software ver.18.Results: The frequency of alleles CYP2C19*1/*1, CYP2C19*1/*17 and CYP2C19*17/*17 were estimated as 58.33, 29.1 and 11.1%, respectively. Specificity and sensitivity of HRM Method were 90% and 100%, with respect to PCR-RFLP. Also, HRM analysis has been evaluated as a faster and more effective approach.Conclusion: Comparison of our results based on HRM analysis with PCR-RFLP showed that our developed Method is rapid, accurate, fast and economic to study the CYP2C19*17 allele and it is appropriate for other similar population genetic studies.

Background: Apolipoprotein E 􁈺, ApoE􁈻,is one of the several important lipoproteins in lipid metabolism, and many studies have been conducted on it. In various studies, the role of this protein in diseases such as Alzheimer's disease, cardiovascular disease, diabetes and immune deficiency diseases such as AIDS have been addressed. The gene is a polymorphic apolipoprotein with three transcripts or different alleles called e2, e3 and e4. These three alleles have six genotypes E2E2, E2E3, E2E4, E3E3, E3E4 and E4E4 with different frequency in the population. Depending on the demographic, geographic and racial characteristics of these genotypes, they are seen with varying frequency in the population, which is in any case the highest frequency related to E3E3. In this study determinated apolipoprotein E Genotyping in diabetic patients with cardiovascular diseases with allele specific PCR Method. Materials and Methods: This study was performed on 107 blood samples of diabetic patients with cardiovascular disease living in Kashan. To determine the genotype of this gene, allele specific PCR with five primers was used and then the pattern of bands in electrophoresis was used. Results: The results show that in this population (diabetic patients with cardiovascular disease living in Kashan), as in most Iranian and world populations, the most genotype is E3E3. Also, the frequency of e2, e3 and e4 alleles is 5. 1, 79 and 15. 9%, respectively. Conclusion: This study shows that there is an average impact of this genotype on lipoproteins of plasma or types of CVD, so it can be helpful to predict and prevent diseases associated with this genotype.