Showing 9 results for Polymerase Chain Reaction
Hossein Goudarzi, Hanieh Rezaee, Mitra Rafizadeh, Elnaz Mirsamadi, Afsoon Mirsamadi,
Volume 15, Issue 5 (10-2012)
Abstract
Background: H.pylori is one of the most common chronic bacterial infections in population so more than 85 percent are infected in Iran. H.pylori can cause different gastrointestinal disease like gastritis, peptic ulcers and even cancer. One of the effective factors in pathogenesis of bacteria is cytotoxin associated with gene A (cagA). Strains with cagA gene are more virulent. The aim of this study was to determine the frequency of cagA gene of H.pylori in patients with gastric disorders who were admitted to Imam Hossein Hospital, Tehran. Materials and Methods: In this descriptive study, DNA was extracted from 84 paraffin- embedded tissues using QiaAmp tissue kit. H.pylori was verified with PCR of 16sRNA sequences specific for Helicobacter spices and cagA gene was determined using specific primer by the PCR method. The prevalence of cagA gene in three clinical groups gastritis, gastric ulcer, and atrophic patients was compared. Results: Among 84 H.pylori positive isolates ,72 biopsy samples were positive for 16sRNA (85.7%) and 46 (63.9%) for cagA. The prevalence of cagA positive strains in peptic ulcer patients (43.5%) was greater than in those with gastritis (30%). Conclusion: Results showed that Helicobacter pylori strains with cagA are more common in patients with peptic ulcer and cancer.
Somayeh Kiaie, Hamid Abtahi, Mohammad Alikhani, Ghassem Mosayebi,
Volume 15, Issue 7 (12-2012)
Abstract
Background: Vibrio cholerae is a gram-negative bacterial pathogen that causes diarrheal disease cholera. One of the most pathogenic factors of Vibrio cholera is pili. Pili plays an important role in colonization and persistence of bacteria in small intestine. Materials and Methods: In this study, pili A (tcpA) gene was amplified by Polymerase chain reaction (PCR) method and sub-cloned into expression vectors such as pGEX4T-1. Escherichia coli competent cells were transformed by recombinant plasmids and the expression of protein with IPTG. The recombinant proteins were purified by affinity chromatography (GST) and immunoblot analysis was used for evaluation of new recombinant proteins antigenicity. The concentration of recombinant proteins was measured according to Bradford assay. Results: The results of this study indicated that recombinant proteins were expressed successfully in competent cell of E. coli, such as E. coli BL21 (DH3). The recombinant protein was purified by affinity chromatography (GST). The immunoreactivity pattern of anti-Tcp antibody with recombinant proteins of TcPA showed that the recombinant proteins had antigenic properties. Conclusion: Because these recombinant proteins are antigenic, these proteins may be considered as tentative candidates for designing cholera vaccine.
Esmaiel Saberfar, Zahra Goodarzi, Ali Najafi,
Volume 15, Issue 8 (1-2013)
Abstract
Background: Influenza type A virus is one of the most important viral agents in human respiratory diseases. The genetic variability of the influenza viruses leads to the incidence of new epidemics worldwide. Hence, there is a growing need for rapid and effective new methods capable of detection and differentiation of influenza virus circulating strains. This study was done to develop a method for rapid differentiation of the subtypes of influenza type A virus. Materials and Methods: In this experimental study, reverse-transcription and polymerase chain reaction (RT-PCR) were performed using a primer set based on M gene of H1N1, H3N2, H5N1, and H9N2 influenza subtypes. Then the amplified fragments were subjected to digestion using subtype specific restriction endonuclease enzymes. Results: The results of PCR reaction showed that the primer pair of the M gene was specific and capable of amplifying all influenza subtypes understudy. Also, different restriction fragment length polymorphism patterns (RFLP) were generated using enzyme digestion reaction on the amplified segment of M gene. Conclusion: RT-PCR and RFLP analysis of the M gene can be employed as a useful method for differentiating influenza virus subtypes
Azar Jafari, Sharbanuo Parchami Barjui, Somaye Reiisi, Morteza Hashemzadeh Chaleshtori, Sepideh Miraj,
Volume 16, Issue 10 (1-2014)
Abstract
Background: Preeclampsia (PE) is a serious problem of pregnancy and its etiology is still unknown. The inheritance of preeclampsia is one of the theories regarding to the etiology of preeclampsia. Methylenetetrahydrofolatereductase (MTHFR) is a key enzyme in folate metabolism and the C677T polymorphism of the MTHFR gene is associated with decrease MTHFR activity, and therefore cause higher blood levels of homocysteine and leads to vascular disease that can be the reason of preeclampsia. The aim of this study was to evaluate the relationship between MTHFR gene C677T polymorphism with PE development in south-west of Iran.
Materials and Methods: This case-control study was performed in 129 preeclamptic pregnant women and 125 control individuals.The C677T polymorphism of the MTHFR gene was determined by PCR-RFLP method.
Results: The CC, CT and TT genotypes frequency of C677T polymorphism of MTHFR gene were 57.4, 38.8 and 3.9 percent in preeclamptic women and 53.6, 40 and 6.4 percent in control group. They were not significantly different (p=0.614). However, the frequency of TT genotype was higher in control group (p=0.36). There was not any significant difference in T allele distribution between preeclamptic women (23.3%) and control group (26.4%).
Conclusion: Our results showed that there was not any correlation between the C677T polymorphism and PE but the TT genotype of C677T polymorphism seems to be a protective factor for preeclampsia.
Mana Shojapour, Ghasem Mosayebi, Fardin Faraji, Keyvan Faraji, Ali Ghazavi,
Volume 17, Issue 3 (6-2014)
Abstract
Background: Multiple sclerosis (MS) is an autoimmune disorder with unknown etiology. Genetic and environmental factors associated with MS susceptibility. Genetic studies show an important role for human leukocyte antigens (HLA) and susceptibility to autoimmune diseases such as MS. There is controversy between the association of HLA alleles with MS susceptibility in various studies. However, with consider the high incidence of MS in Iranian population and limit information about association of HLA and MS, we analyzed HLA alleles in MS patients.
Materials and Methods: In this case-control study, 60 MS patients and 40 normal individuals with the same ethic background and geographic area were analyzed for HLA-DRB and DQB alleles by single specific primer-polymerase chain reaction (SSP-PCR) method.
Results: HLA-DRB1*03 and DQB1*02 alleles frequencies in MS patients were greater than healthy controls. There was no significant difference in frequency of other HLA-DR alleles between the MS patients and normal individuals.
Conclusion: DRB1*03 and DQB1*02 alleles confer increased susceptibility to MS in this population. However, to determine the role of HLA in Iranian MS patients, more studies are needed.
Meysam Hasan Nejad Bibalan, Ezzatollah Ghaemi, Fateme Shakeri, Naeme Javid,
Volume 17, Issue 6 (9-2014)
Abstract
Background: Staphylococcus aureus is a gram-positive bacterium that has remained a persistent pathogen, causing infections such as endocarditis and toxic shock syndrome in humans. The accessory gene regulator (agr) system of Staphylococcus aureus is responsible for controlling the expression of many genes that code virulence factors and hemolysis.This study was carried out to determine the S.aureus agr group based on their source of isolation and any relation between agr specificity groups, pigmentation and hemolysis .
Materials and Methods: DNA of 194 S. aureus isolates were extracted by lysozym-phenol chloroform method, included 85clinical samples, 58 samples which isolated from nose of health care workers and 51 cases obtained from food product in Gorgan, North of Iran. PCR-based assays were used to evaluate agr locus nucleotide polymorphism for the identification of agr specificity group. Pigmentation on nutrient agar medium and hemolysis on sheep Blood agar medium were assessed.
Results: The majority of isolates belonged to agr group I (43.3%), followed by agr group III (28.87%), agr group II (22.68%), and agr group IV (5.15%). The isolates belonged to agr group IV have greater ability to produce hemolysin (60%) whereas isolates belonged to agr group III have greater ability to produce pigment (60.5%).
Conclusion: agr group I was predominant among health care worker and food product specimens in Gorgan, North of Iran but in strains isolated from patient, agr group III was predominant. Investigation of the possible role of agr group III in Staphylococcus aureus infection in the next studies is recommended.
Somayyeh Moatti, Behrouz Shojaee Sadi, Ehsanollah Ghaznavi-Rad,
Volume 20, Issue 7 (10-2017)
Abstract
Abstract
Background: Integrons are mobile genetic elements that play an important role in dissemination of antibiotic resistance genes. The aim of present study is to determine the antibiotic resistance profile, frequency of integrons genes (class 1, 2, 3) and compare it between MRSA and MSSA isolates from clinical infections.
Materials and Methods: 50 MRSA and 50 MSSA isolates from March to September 2015 were isolated from infection site of hospitalized patients referred to Valiasr hospital Arak, Iran were subjected to this study. All isolates were tested for susceptibility to antibiotics using disk diffusion method. Then, the mecA gene was studied to validate resistance. The frequency of integrons (class 1, 2, 3) and the variable region genes like qacE
1 and sul1 in isolates were determined by PCR method.
Results: The highest antibiotic resistances rate in isolates was found for clindamycin.
All of the isolates were susceptibel to vancomycin. 80% of MRSA and 40% of the MSSA isolates carried class 1 integrons, whereas class 2 integron were found in 12 % and 4% of MRSA and MSSA isolates, respectively. Also, all isolates that were class 1 integron gene positive contain qacE1 and sul1 genes. Class 3 integrons were not found.
Conclusion: The high frequency of class 1 integron in MRSA and MSSA isolates associated with high rate of antibiotic resistance indicating that may be integrons play an important role facilitating the spread of antimicrobial resistance in this region. Clinical doctors and infection control committee should take this issue seriously.
Mehrdad Nasrollahzadeh Sabet, Mohammad Foad Heidari, Mohammad Khanalipour, Saadat Allah Ghaffari, Milad Jafari Ashiani, Sajjad Biglari, Emran Esmaeilzadeh,
Volume 23, Issue 5 (11-2020)
Abstract
Background and Aim: Since late 2019, with the emergence of a new type of coronavirus that causes a new respiratory disease called COVID-19, there have been many concerns about the spread of this disease and how to deal with it. Due to the ability of the virus to be transmitted rapidly, diagnosing the infected individuals in the early stages for isolating them is critical. This study aims to evaluate the reliability of Computed Tomography (CT) scan in diagnosing COVID-19.
Methods & Materials: Participants were 212 patients admitted to hospital with confirmed diagnosis of COVID-19. Demographic information, medical history, symptoms, and the chest CT scan results were collected and analyzed. Finally, the power of CT scans in the diagnosis of this disease was compared with the Real-Time Polymerase Chain Reaction (RT-PCR) molecular test.
Ethical Considerations: This study received ethical approval from the ethics committee of AJA University of Medical Sciences (Code: IR.AJAUMS.REC.1399.091).
Results: The sensitivity of CT scan in the diagnosis of COVID-19 was relatively high, but its false-positive results were also high.
Conclusion: CT scan is a relatively sensitive method for diagnosing COVID-19, but caution should be made due to its high false-positive results which can lead to increased financial burden on the health system.
Ahmad Hamta, Sahar Adl,
Volume 24, Issue 1 (3-2021)
Abstract
Background and Aim: Breast cancer is the most common cancer type and the leading cause of cancer-induced deaths in women, worldwide. The Fibroblast Growth Factor Receptor 2 (FGFR2) is a tyrosine kinase receptor that plays an essential role in the growth, invasion, movement, and angiogenesis of tumor cells. Several single nucleotide polymorphisms have been found in the intron 2 of the FGFR2 gene, i.e., associated with a high risk of breast cancer. Genetic variation in this receptor is a new risk factor for breast cancer. The current study aimed to evaluate the association of single-nucleotide polymorphism rs2981582C/T in women with breast cancer.
Methods & Materials: In total, 80 women with breast cancer and 80 healthy women (controls) were selected from Markazi Province, Iran to participate in this research. Polymorphism rs2981582 was analyzed to investigate its association with breast cancer. DNA extraction from blood samples was performed using a kit. The presence of these single-nucleotide polymorphisms was determined by Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR - RFLP). Statistical analyses were performed by SPSS using Chi-squared test at P≤0.05.
Ethical Considerations: This study was approved by the Ethics Committee of the Arak University (Code: IR.ARAKMU.REC.1395.28).
Results: Significant differences were observed in the frequency of rs2981582 polymorphism in the FGFR2 gene between the control and patient groups (P=0.000). In the patient group, the TT genotype was significantly associated with the risk of breast cancer (P=0.001; OR=3.566). On the other hand, allele C indicated a protective role against the disease (P=0.000).
Conclusion: The obtained data revealed a significant relationship between rs2981582 C/T polymorphism and the risk of breast cancer; thus, this single-nucleotide polymorphism could be used as a biomarker to predict breast cancer.
