Microbiology
Samaneh Abolbashari; MohammadTaghi Shakeri; Maryam Hami; Aida Gholoobi; Amin Hooshyar Chechaklou; Mohammad Sadegh Damavandi; Aref Movaqar; Razieh Yousefi; Zahra Meshkat; Saeedeh Hajebi Khaniki
Abstract
Background & Objective: Polyomaviruses types BK and JC and Cytomegalovirus (CMV) have been shown to be related to kidney transplantation complications. This study aimed to assess the prevalence of these viruses in patients receiving kidney transplantation.Methods: This cross-sectional study was performed ...
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Background & Objective: Polyomaviruses types BK and JC and Cytomegalovirus (CMV) have been shown to be related to kidney transplantation complications. This study aimed to assess the prevalence of these viruses in patients receiving kidney transplantation.Methods: This cross-sectional study was performed on 40 kidney transplant recipients and 44 donors. Urine samples were used for the extraction of viral DNA. The prevalence of JC and BK viruses and their viral loads were determined by real-time polymerase chain reaction.Results: JC and BK viruses were identified in 31% and 92.3% of all subjects, respectively. The frequency of JC and BK cases was not statistically different between the recipient and donor groups (P>0.05). All patients in the donor group and 96.8% of the recipients were positive for CMV IgG antibody. The mean viral load of BK in donors and recipients was 4.5×1010 and 3.3×1011 copies, respectively. The mean viral load of JC was 8.6×107 copies in donors and 2.9×108 copies in recipients. The distribution of BKV was significantly higher in recipients than donors (P=0.001), while no difference was observed between the two studied groups for JCV.Conclusion: This study showed a relatively high prevalence of BK and JC viruria in both renal transplant donors and recipients. The viral load for BKV, but not JCV, was higher in recipients than in donors.
Microbiology
Faria Hasanzadeh Haghighi; Ehsan Aryan; Mohammad Derakhshan; Aida Gholoobi; Zahra Meshkat
Volume 13, Issue 4 , October 2018, , Pages 403-407
Abstract
Background & objective: Tuberculosis (TB) remains a major cause of death around the world. Bacillus Calmette Guérin (BCG) is the only vaccine used in TB prevention that has a protective effect in children, but its effectiveness declines in adults. Design and development of new vaccines ...
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Background & objective: Tuberculosis (TB) remains a major cause of death around the world. Bacillus Calmette Guérin (BCG) is the only vaccine used in TB prevention that has a protective effect in children, but its effectiveness declines in adults. Design and development of new vaccines is the most effective way against TB. The aim of this study was to design and construct a DNA vaccine encoding mtb32C and mpt51 fusion genes of Mycobacterium tuberculosis. Methods: First, mpt51 fragment was amplified by PCR method. The pcDNA3.1+/mtb32C plasmid was transformed into E. coli JM109 and then extracted. The mpt51 gene and pcDNA3.1+/mtb32C plasmid were both digested with EcoRI and BamHI restriction enzymes followed by ligation of mpt51 fragment into the digested vector. The recombinant plasmid containing mtb32C and mpt51 was subsequently transformed into competent E. coli TOP10 strain. The clones were confirmed by colony-PCR, restriction enzyme digestion and sequencing. Results: Using agarose gel electrophoresis, a 926 bp fragment corresponded to mpt51 was observed. Digestion of the vector pcDNa3.1+/mtb32C and mpt51 gene was confirmed by electrophoresis. Then, the pcDNA3.1+/mtb32C plasmid was extracted. Sequencing results confirmed the accuracy of the desired plasmid. Conclusion: In this study, we constructed a cloning vector encoding Mtb32C/Mpt51 gene of Mycobacterium tuberculosis. The eukaryotic expression of this vector can be confirmed in future studies. It can be considered as a DNA vaccine in animal models later. Successful cloning provides a basis for the development of new DNA vaccines against tuberculosis.
Amir Tajbakhsh; Faezeh Ghasemi; Seyedeh Zohre Mirbagheri; Mastoureh Momen Heravi; Mehdi Rezaee; Zahra Meshkat
Volume 13, Issue 4 , October 2018, , Pages 429-437
Abstract
Background and Objectives: The incidence of rifampin-resistant strains of Mycobacterium tuberculosis has attracted more attention than the tuberculosis infection due to laborious treatment and control. Recognizing the Mycobacterium tuberculosis genotypes involving in drug resistance via multiplex PCR, ...
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Background and Objectives: The incidence of rifampin-resistant strains of Mycobacterium tuberculosis has attracted more attention than the tuberculosis infection due to laborious treatment and control. Recognizing the Mycobacterium tuberculosis genotypes involving in drug resistance via multiplex PCR, a simple and rapid genotyping method, is an emergency for better treatment and control of tuberculosis. This study was designed to specify the frequency of rifampin-resistant strains of Mycobacterium tuberculosis isolated from patients by multiplex allele-specific Polymerase Chain Reaction assay (MAS-PCR).Methods: In this study, 88 Mycobacterium tuberculosis positive samples were included from Qaem Hospital, Mashhad. MAS-PCR was used to detect the rifampin resistance associated mutations in rpoB gene. Results: Mutations in three codons of rpoB gene causing rifampin resistance were detected in 51 isolates (58.96%). The detected mutations in codons 531, 526, and 516 were 55.68%, 38.63%, and 13.63%, respectively. The simultaneous mutations were detected in 11 isolates (12.50%) in codons 531, 526 and 516, in 21 isolates (23.86%) in codons 531 and 526, and in one isolate (1.13%) in codons 526 and 516. Conclusion: According to the results of this study, the frequency of rifampin-resistant strains of Mycobacterium tuberculosis isolated from Khorasan province patients (North-East of Iran) was high. The developed MAS-PCR assay can be used for rapid detection in clinical diagnostic laboratories in areas with high prevalence of multidrug-resistant Mycobacterium tuberculosis strains. In this respect, MAS-PCR is simple, rapid, and highly sensitive method for drug susceptibility tests for detecting multidrug-resistant Mycobacterium tuberculosis.
Microbiology
Faezeh Ghasemi; Majid Ghayour Mobarhan; Hamed Gouklani; Zahra Meshkat
Abstract
Hepatitis C virus (HCV) is responsible for a vast majority of liver failure cases. HCV is a kind of blood disease appraised to chronically infect 3% of the worlds’ population causing significant morbidity and mortality. Therefore, a complete knowledge of humoral responses against HCV, resulting ...
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Hepatitis C virus (HCV) is responsible for a vast majority of liver failure cases. HCV is a kind of blood disease appraised to chronically infect 3% of the worlds’ population causing significant morbidity and mortality. Therefore, a complete knowledge of humoral responses against HCV, resulting antibodies, and virus-receptor and virus-antibody interactions, are essential to design a vaccine. HCV epitopes or full sequence of HCV proteins can induce HCV specific immune responses. In fact, structural proteins are usually the main target of humoral responses and non-structural proteins are usually the main target of cellular responses. Hence, various vaccines based on distinct antigenic combinations are developed to prevent HCV infection and the current study tried to summarize them.
Microbiology
Nafiseh Izadi; Mahboubeh Naderi Nasab; Elnaz Harifi Mood; Zahra Meshkat
Abstract
Background and Objectives: Since the fluoroquinolones are the broad-spectrum antibiotics, they affect both Gram-negative and Gram-positive bacteria. These antibiotics are widely prescribed by physicians. As a result, some bacteria, especially Enterobacteriaceae, have shown a resistance to this family ...
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Background and Objectives: Since the fluoroquinolones are the broad-spectrum antibiotics, they affect both Gram-negative and Gram-positive bacteria. These antibiotics are widely prescribed by physicians. As a result, some bacteria, especially Enterobacteriaceae, have shown a resistance to this family of antibiotics. The current study aimed at detecting the frequency of qnrA, qnrB, and qnrS genes, novel plasmid-mediated quinolone-resistance genes, among extended-spectrum β-lactamases (ESBL)-positive and ESBL-negative Klebsiella pneumoniae isolates. Materials and Methods: One hundred and thirty isolates of K. pneumoniae were collected from Imam Reza Hospital and its associated clinics from May 2011 to July 2012. The isolates were tested for ESBLs by the conventional methods. Polymerase chain reaction (PCR) was performed to amplify qnr A, B, and S. Results: Thirty-eight (29.3%) isolates were ciprofloxacin-resistant. Among 130 K. pneumoniae infectious isolates, 56 (43%) were capable of producing ESBL; 10.8% (n=14), 15.4% (n=20), and 20.8% (n=27) of ESBL-producing K. pneumonia were positive for qnrA, qnrS, and qnrB, respectively, and 13.8% (n=18) of the isolates harbored 2 or 3 qnr genes. Conclusion: The results of the current study showed that quinolone-resistance genes were more frequent in ESBL-producing K. pneumoniae (37.5%) isolates, compared with the ESBL-negative isolates (20.89%). The prevalence of qnr genes was high in K. pneumoniae isolates, with higher frequency in ESBL-positive strains. Most of the isolates were positive for all 3 groups of qnr genes and the qnrB was the most common one.
Microbiology
Sina Rostami; Alireza Pasdar; Sina Gerayli; Hamed Hatami; Samaneh Sepahi; Fatemeh Nategh; Mojtaba Meshkat; Seyed Mousalreza Hoseini; Mitra Ahadi; Hamid Reza Sima; Hasan Vosughinia; Mohammad Reza Sarvghad; Abbas Esmaeelzade; Hosein Nomani; Homan Mosanan Mozafari; Fariba Rezai Talab; Mohammad Taghi Shakeri; Zahra Meshkat
Abstract
Background and Objectives: Interferon-gamma is an important cytokine, which facilitates immunity against intracellular pathogens. Several factors, including genetic variations of cytokine-producing genes have been shown to influence the progression and severity of Hepatitis C virus (HCV) infection. Methods: ...
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Background and Objectives: Interferon-gamma is an important cytokine, which facilitates immunity against intracellular pathogens. Several factors, including genetic variations of cytokine-producing genes have been shown to influence the progression and severity of Hepatitis C virus (HCV) infection. Methods: Between January and December 2012, 87 HCV-infected individuals and 89 individuals without HCV infection were recruited for the study of Single Nucleotide Polymorphism (SNP) at Interferon Gamma (IFNG) +874 T/A. After extraction of genomic DNA from Peripheral Blood Mononuclear Cells (PBMCs) in blood sample of the individuals, Amplification Refractory Mutation System (ARMS) polymerase chain reaction was performed to evaluate the SNP at this position. Results: The frequency of genotype TA was 62.1% in the HCV-infected group, while it was 47.2% for the control group (p=0.033). However, after adjusting for confounders (including alcohol consumption, drug addiction, transfusion, and tattoos), the genotypes at this position did not show any statistically significant association with HCV infection (adjusted P values were above 0.05). The frequency of allele A was slightly higher in patients than the controls (55.2% versus 48.3%).Carriers of A allele were more frequent in patients with HCV infection compared to the control group (55.17% in patients versus 48.31% in the control group; P=0.02). However, after adjustment for confounders, the results were no longer statistically significant (P=0.2). Conclusion: A carrier status for certain alleles and genotypes at Interferon Gamma (IFNG) +874 T/A may lead to higher susceptibility to HCV infection in a certain population.
Microbiology
Roghayeh Teimourpour; Amineh Sadat Tajani; Vahid Reza Askari; Sina Rostami; Zahra Meshkat
Volume 11, Issue 3 , July 2016, , Pages 222-230
Abstract
Background: Hepatitis C virus (HCV) infection is one of the most prevalent infectious diseases responsible for high morbidity and mortality worldwide. Therefore, designing new and effective therapeutics is of great importance. The aim of the current study was to construct a DNA vaccine containing ...
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Background: Hepatitis C virus (HCV) infection is one of the most prevalent infectious diseases responsible for high morbidity and mortality worldwide. Therefore, designing new and effective therapeutics is of great importance. The aim of the current study was to construct a DNA vaccine containing structural proteins of HCV and evaluation of its expression in a eukaryotic system. Methods: Structural proteins of HCV (core, E1, and E2) were isolated and amplified from JFH strain of HCV genotype 2a using PCR method. The PCR products were cloned into pCDNA3.1 (+) vector and finally were confirmed by restriction enzyme analysis and sequencing. The eukaryotic expression of the vector was confirmed by RT-PCR. Results: Recombinant vector containing 2241bp fragment of HCV structural genes was constructed.The desired plasmid was sequenced and corresponded to 100% identity with the submitted sequences in GenBank. RT-PCR results indicated that the recombinant plasmid could be expressed efficiently in the eukaryotic expression system. Conclusion: Successful cloning of structural viral genes in pCDNA3.1 (+) vector and their expression in a eukaryotic expression system facilitates the development of new DNA vaccines against HCV. A DNA vaccine encoding core-E1-E2 antigens was designed. The desired expression vector can be used for further attempts in the development of vaccines.
Microbiology
Samira Rashidian; Roghayeh Teimourpour; Zahra Meshkat
Volume 11, Issue 2 , April 2016, , Pages 112-119
Abstract
Background: Tuberculosis (TB) remains as a major cause of death around the world. Construction of a new vaccine against tuberculosis is an effective way to control it. Several vaccines against this disease have been developed. The aim of the present study was to cloning of tb10.4 gene in pcDNA3.1+ plasmid ...
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Background: Tuberculosis (TB) remains as a major cause of death around the world. Construction of a new vaccine against tuberculosis is an effective way to control it. Several vaccines against this disease have been developed. The aim of the present study was to cloning of tb10.4 gene in pcDNA3.1+ plasmid and evaluation of its expression in eukaryotic cells. Methods: Firstly, tb10.4 fragment was amplified by PCR and the PCR product was digested with restriction enzymes. Next, it was cloned into pcDNA3.1+ plasmid. Following that, pcDNA3.1+/tb10.4 recombinant plasmid was transfected into eukaryotic cells. Results: 5700 bp band for pcDNA3.1+/tb10.4 recombinant plasmid and 297 bp fragment for tb10.4 were observed. Cloning and transfection were successful and designed recombinant vector was confirmed by sequencing. Conclusion: Successful cloning provides a basis for the development of new DNA vaccines against tuberculosis. In the current study, the aim was cloning of tb10.4 gene in pcDNA3.1+ plasmid and transfection into eukaryotic cells.
Microbiology
Faezeh Ghasemi; Sina Rostami; Maryam Sadat Nabavinia; Zahra Meshkat
Volume 11, Issue 1 , January 2016, , Pages 41-46
Abstract
Background: Human papillomavirus (HPV) is responsible for the development of cervical neoplasia. Infection with human papillomavirus type 16 (HPV-16) is a major risk factor for the development of cervical cancer. The virus encodes three oncoproteins (E5, E6 and E7), of which, the E7 oncoprotein ...
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Background: Human papillomavirus (HPV) is responsible for the development of cervical neoplasia. Infection with human papillomavirus type 16 (HPV-16) is a major risk factor for the development of cervical cancer. The virus encodes three oncoproteins (E5, E6 and E7), of which, the E7 oncoprotein is the major protein involved in cell immortalization and transformation of the infected cells. The aim of the current study was to develop Michigan Cancer Foundation 7 (MCF7) cells, which could stably express E7 protein of HPV type 16. Methods: E7 gene of HPV type 16 was introduced into MCF7 cells by Lipofectamine 2000 reagent and the transfected cells were treated with G418 antibiotic. After antibiotic selection of the transfected cells, stable expression of E7 gene of HPV16 was confirmed by reverse transcriptase polymerase chain reaction (RT-PCR). Results: Antibiotic selections of transfected cells were performed and transfected cells were alive in cytotoxic concentration of the antibiotic. RNA was extracted from transfected cells and E7 gene of HPV16 was amplified by RT-PCR method and a 350-bp band corresponds to E7 was observed. Conclusion: Results confirmed the stable transfection of cells. The stably transfected cells can be used as a useful tool in future studies on HPV16 and cancers caused by this virus.
Masoumeh Salehpour; Naser Tayyebi Meibodi; Roghayeh Teimourpour; Adel Ghorani-Azam; Samaneh Sepahi; Sina Rostami; Zahra Meshkat
Abstract
Background &Objective: Breast cancer is the most common female malignancy. Detection of DNA of human papillomaviruses (HPVs) in breast carcinomas suggests that the virus may play a role in the pathogenesis of this disease. The aim of this study was to evaluate the frequency of HPVs genotypes 6, 11, ...
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Background &Objective: Breast cancer is the most common female malignancy. Detection of DNA of human papillomaviruses (HPVs) in breast carcinomas suggests that the virus may play a role in the pathogenesis of this disease. The aim of this study was to evaluate the frequency of HPVs genotypes 6, 11, 16, 18 and 31 in paraffin-embedded tissue samples of invasive breast carcinomas. Methods: Three hundred and twenty six paraffin-embedded tissue samples of breast cancer were studied. PCR was performed using specific primers for HPV genotypes. Results: Of total 206 (63.2%) samples positive for Beta-globin gene, 54 (26.2%) were HPV-positive and 152 (73.8%) were negative for HPV. Distribution of HPV genotypes were as follows: 19 (25.7%) were positive for genotype 11, 5 (6.8%) were positive for genotype 6; and 2 cases (2.7%) were positive for both genotypes 6 and 11. Samples were also screened for HPV genotypes 16, 18 and 31 but none was positive. Conclusion: The current study confirmed the association of HPV and breast cancer. However, all samples were negative for high-risk HPV types 16, 18 and 31. How to cite this article: Salehpour M, Tayyebi Meibodi N, Teimourpour R, Ghorani-Azam A, Sepahi S, Rostami S, et al. Frequency of Human Papillomavirus Genotypes 6, 11, 16, 18 And 31 in Paraffin-Embedded Tissue Samples of Invasive Breast Carcinoma, North-East of Iran. Iran J Pathol. 2015;10(3):192-8.
Nafiseh Izadi; Mahboubeh Naderi Nasab; Elnaz Harifi Mood; Zahra Meshkat
Volume 9, Issue 3 , July 2014, , Pages 199-205
Abstract
Background & Objectives:Extended-spectrum-B-lactamase (ESBL)-producing strains of Klebsiella Pneumoniaare an important cause of many serious infections in hospitalized and nonhospitalized patients and delayed treatment of these infections in crease chance of death in patients. This study was performed ...
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Background & Objectives:Extended-spectrum-B-lactamase (ESBL)-producing strains of Klebsiella Pneumoniaare an important cause of many serious infections in hospitalized and nonhospitalized patients and delayed treatment of these infections in crease chance of death in patients. This study was performed to determine the prevalence of ESBL-producing K. Pneumonia and to evaluate the frequency of TEM and SHV genes among the clinical samples. Methods:One hundred and thirty isolates of K. Pneumoniawere collected at Imam Reza Hospital in Mashhad (Iran) from May 2011 to July 2012. ESBL production was determined by the double disk diffusion (DDs) test. PCR method was used to detect TEM and SHV genes. Results:Of 130 patients withK. pneumoniainfection 28 were out-patients and 102 hospitalized patients. The most specimens was urine samples (n=25 in out-patients, n=39 in hospitalized patients, totally 49.2%) followed by wound samples (n=3 in out-patients, n=21 in hospitalized patients, totally 21.5%), blood samples (n=19 in hospitalized, 14.6%). The prevalence of ESBL producingK. pnemoniaewas estimated 43% (n=56) including three of ESBLs positive isolates from out-patients and 53 from hospitalized patients. Of 56 ESBLs positive isolates, 44(87.54%) TEM, 39(69.64%) SHV and in 27 cases (48.21%) both TEM and SHV were detected. Conclusion:A high prevalence of ESBL-producing K. Pneumoniaamong the hospital isolates obtained of urinary followed by blood and wound samples were documented. The majority of them carried both TEM and SHV genes. Results of this study alarm for the physicians because treatment and control nosocomial infections for them were difficult.
Reza Akhavan; Zahra Meshkat; Mehrangiz Khajekaramadini; Mojtaba Meshkat
Volume 8, Issue 2 , April 2013, , Pages 73-80
Abstract
Background & Objectives: Tuberculosis is one of the greatest health problems in Iran. The distribution of the disease is not equal in all parts of the country. The aim of this study was to evaluate the frequency of positive results for Mycobacterium tuberculosis in samples referred to an academic ...
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Background & Objectives: Tuberculosis is one of the greatest health problems in Iran. The distribution of the disease is not equal in all parts of the country. The aim of this study was to evaluate the frequency of positive results for Mycobacterium tuberculosis in samples referred to an academic hospital in an 8 year period.
Materials and Methods: The samples from different wards of Qaem Hospital, Mashhad and samples referred to Outpatient Clinic during the years 2001-2008 and 75 samples from the prison in the same period were analyzed with direct microscopy of smear and culture methods for M. tuberculosis. Basic descriptive statistics were performed using SPSS 11.5 software.
Results: A total 26817 samples were analyzed and the results showed that the frequency of Mycobacterium positive samples in hospitalized patients' samples was 2412 (9%) with microscopy and 1573 (6%) with culture method. In the outpatients, it was 897 (10.2%) and 417 (4.7%) with microscopy and culture methods, respectively. Form 75 samples from the prison, 9 (12%) were positive with microscopy method. Culture method yielded only one (1.3%) positive result in these samples.
Conclusion: The frequency of M. tuberculosis was relatively high in the study groups. Therefore it seems continues surveillance is essential to monitor the M. tuberculosis in hospitals and community.
Zahra Meshkat; Hessam Mirshahabi; Hoorieh Soleimanjahi; Zuhair Mohamad Hassan
Volume 4, Issue 2 , April 2009, , Pages 65-70
Abstract
Background and Objectives: Some of the human papillomaviruses (HPVs) can infect genital tracts and are sometimes associated with anogenital tract cancers. HPVs induced cervical cancers through the expression of E6 and E7 genes by inactivating the tumor suppressor proteins. In this study, E6 and ...
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Background and Objectives: Some of the human papillomaviruses (HPVs) can infect genital tracts and are sometimes associated with anogenital tract cancers. HPVs induced cervical cancers through the expression of E6 and E7 genes by inactivating the tumor suppressor proteins. In this study, E6 and E7 genes were chosen in order to construct an expression vector which is able to express target proteins. Patients and Methods: This experimental investigation was performed in Virology Department of Tarbiat Modares University. An expression vectorcontaining human papillomavirus type 16 E6 and E7 genes was constructed. The accuracy of the plasmid was confirmed by polymerase chain reaction (PCR) and restriction enzyme analysis. The construct was transfected into the eukaryotic cells and its ability for protein production was confirmed by Western blotting. Results:The colonies containing desired plasmid have the fragment about 995 bp. For confirming the ability of the construct for protein production in eukaryotic cells, Western blotting was done using the lyses-cells as antigen and they showed the desired bands using monoclonal antibodies. Conclusion: The designed vector can consider as a based vaccine for construction a therapeutic vaccine in suitable vectors for gene therapy in order to administration in Iranian patients with cervical cancer.
