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Abstract

Background and objectives:

most common causes of morbidity and mortality in industrial and

developing countries. Recent studies have suggested that

A coronary heart disease is one of the

Helicobacter pylori

heart disease therefore, this study was carried out in Gorgan, Iran, to

show the relationship between coronary disease and Helicobacter

pylori infection.

caused infection may be associated with chronic

Materials and Methods:

carried out on 109 patients suffering from acute coronary syndrome

and 85 healthy individuals, ELISA was used to determine Anti

Helicobacter pylori Anti bodies (IgA, IgG ).

In this cross sectional case-control study

Results:

while in control group were %32.9 and %62.4. There was significant

difference between IgA of two groups (p<0.007). Simultaneous

presence of both IgG and IgA in patients affected by Coronary disease

was meaningful (p<0.003).

IgA and IgG antibodies of case group were %51.4 and %53.2

Conclusion

be related to coronary disease, we suggest their investigation in

suspected individuals.

: Since Simultaneous presence of both IgG and IgA may

Key word

Antibody

: Acute Coronary syndrome, Helicobacter Pylori,

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Abstract Background and objectives: Diarrhea is one of the main cases of morbidity and mortality among children in developing countries. Enteroaggregative Escherichia coli (EAggEC) is an emerging diarrheal pathogen that has been associated characteristically with persistent diarrhea among infants, particularly in the developing Counties. Therefore, we decided to study the prevalence of enteroaggregative strain in cases of Diarrhea in Gorgan by PCR method. Material and Methods: This descriptive study was carried out on 455 subjects suffered from Diarrhea in Gorgan during one year (2005-6). At first, the samples were cultivated on the MacConkey agar and EMB agar media, Then all colony Suspected to E.coli were chosen and their DNA extracted by phenol chloroform method. The result was obtained by the selected primer, PCR method. Results: of 455 samples, Twenty cases (4/4%) including men (12) and woman (8) are positive for EAggEC, 85% of sufferers are under 5 years old (45.8% of them are under one year old). The Prevalence of this gene in Summer , Autumn ,Winter ,Spring are 5.3% , 4.2% , 4.1% and 1.8% ,respectively. Conclusion: Based on the prevalence of Enteroaggregative Escherichia coli (EAggEC) in Diarrheagenic cases in Gorgan (4.4%), we do recommend using molecular methods, which are reliable and less expensive than classic methods, in detecting of microorganisms. Key words: Entroaggregative Escherichia coli, Diarrhea, PCR, Gorgan.

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Abstract Background and objectives: With almost nine million new cases each year, tuberculosis is still one of the most Life-threatening diseases in the World. Distribution of drug resistant strains of M.tuberculosis has a lot of importance. This research was carried out to determine the frequency of drug resistance of M. tuberculosis in strains isolated in Golestan province. Material and Methods: In this cross -sectional study, 104 isolate of M.tuberculosis which isolated from patients referred to Gorgan tuberculosis Health Center, in 2008 were studied. DNA was extracted by Boiling Method. By using PCR method, we determine the M.tubeculosis strain and resistance to Rifampin (Using IS6110 and Gene rpoB primers) and resistance to Isoniazid (Using InhA and KatG primers). As a Gold Standandard, “Proportional method” was performed for 45 Samples. Results: 87 strains were identified as M.tuberculosis. 6.9% of them were resistant to Isoniazid, 4.6% to Rifampin and 2.3% to both (MDR).Sensitivity and Specifity of PCR method in detection of resistant to Isoniazid were 95.3% and 57.1% and for Rifampin were 94.7% and 33.3%. Conclusion: We found that in our region, the MDR is not very common. More than 16% of isolated strains from tuberculosis suspected patients were MOTT, for this reason it is necessary to mention that use biochemical or PCR method to determine M.tuberculosis is necessary. Key words: Mycobacterium tuberculosis, MDR, PCR, Proportional method , Golestan province.

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Abstract Background and objectives: After respiratory infection, Diarrhea is the second cause of mortality. Yersinia enterocolitica is the second important cause of infectious diarrhea in children of some countries. In this study, we evaluated the frequency of Yersinia entocolitica of diarrheal specimens in Gorgan, Iran. Material and Methods: This descriptive cross - sectional Study was carried out on diarrheal stools of 455 patients referred to medical centers and laboratory of Gorgan in 2004-2005. DNA extraction using phenol chloroform was performed for all samples. Using two specific primers (genus-specific16s rRNA and ail- specific species genus of Yersinia enterocolitica), we did PCR sample. Results: Yersinia genome was identified in 12 patients(2.63%) and 11 of them was Yersinia enterocolitica. The frequency infection in of girls (3%) was more than boys (2.4%), and the prevalence in winter (4%) was more them other seasons, and under one- year- group (3.4%) and 1-5 years (3.1%) is more than other age groups. It was not observed significant difference. (P> 0.05). Conclusion: The frequency of Yersinia in cases of diarrhea in Gorgan is similar to most regions of Iran and in children under 5 years is observed more in winter. Key words: Yersinia enterocolitica, Diarrhea, children, Gorgan

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Background and Objectives: rapid, accurate and cost effective diagnosis of infectious and non infectious diseases is an essential step for treatment process. Nowadays, in Line with scientific progression in molecular biology, genetics and biochemistry which are based on biotechnology and genetic engineering aspects, new branch of medicine entitled molecular medicine is being derived. It can be helpful in three areas of diagnosis, prophylaxis and treatment. This new branch is going to identify further complexity of diseases and to present efficient solutions for growing health criteria. Therefore, updating and being familiar with the new procedures related to diagnosis, prophylaxis and therapy are necessary for our society. In this paper, we are trying to introduce NASBA technology which has a high potential, at genome level, in recognizing specific characteristic and unique genetic markers of microorganisms. This technology has numerous benefits for easy detection of infectious diseases such as tuberculosis. Furthermore, we review the methods of tuberculosis detection.

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Abstract Background and objectives: Mumps virus is one of the first known causative agents of meningitis in children. On-time diagnosis is the first step in treating meningitis. We aimed to evaluate Mumps virus meningitis in children in Gorgan, Iran Material and Methods: CSF and blood samples were taken from children with meningitis, Jun 2008 till Sep 2010. For 40 samples with negative bacterial culture, Extraction of viral RNA was carried out and Real-time PCR was performed for detection of Mumps virus. Demographic, clinical, biochemical and cytological data were collected. We run SPSS version 18 to analyze the data, using Chi Square (p<0.05). Results: three (7.5 %) samples have Mumps virus, two boys and one girl. All three positive cases have 0.5-1 degrees Celsius fever and vomiting but no bulging fontanel. They have not Kernig, Rodor, Brudzinski’s sign, hepatosplenomegaly, lymphadenopathy, pharyngitis and rash. ESR is higher than normal in all positive cases and CRP is positive in two cases. Protein of CSF in one case is higher than normal range. Conclusion: meningitis is an emergency condition therefore, molecular diagnostic techniques are recommended for early diagnosis and intervention. Key words: meningitis, mumps virus, cerebrospinal fluid, Real-Time PCR

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Abstract Background and objectives: The increase of ESBL producing E.coli can create a tremendous difficult y for the health system. These isolates leads to rapid transmission of causative genes to other clinically important bacteria and synchronously increased resistant to other antibiotics. This study was carried out to determine the prevalence of this isolate and related genes in Gorgan, North of Iran. Material and Methods: This study was conducted on 218 isolated E.coli from urinary tract infection of outpatients referring to six medical laboratories in Gorgan, during 2010-11. The resistance to Cefotaxim (Mast Co.) was assessed by Kirby-Bauer disk diffusion method. The confirmatory test for detection of resistant isolates was carried out by double disk method at the presence of Cefotaxim and clavulanic acid. The presence of β lactamase gene of blatem, blactx and blashv in ESBL was assessed by PCR method. Results: of 218, 70 isolates (32.1%) are resistant to Cefotaxim. Sixty-two (88.6%) of them are confirmed as ESBL producing E.coli. β lactamase genes of blactx, blatem and blashv can be seen in 28(45.2%), 26(41.9%) and 6(9.7%) isolates, respectively. Conclusion: the prevalence of ESBL producing E.coli in Gorgan is in the range of country average and blaCTX-M gene is the most common gene. Key words: E.coli, ESBL, bla gene, UTI

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Abstract Background & Objective: Hepatitis E virus is one of the most common causes of acute infection in adults. Pregnant and transplant patients are more in risk of HEV infection. Fecal-oral is the main route of HEV transmission but recently transmission by blood transfusion has been observed. This study was aimed to determine the prevalence of HEV-Ab in hemodialysis patients in Gorgan, Iran. Material and Methods: In this cross-sectional descriptive study, we investigated 150 hemodialysis patients of Panje Azar hospital in Gorgan. These patients were evaluated for the presence of HEV total Ab by ELISA method. Results: of 150, 6 patients (4%) are positive for HEV-Ab. There has been no significant relation between anti HEV Ab and variables such as age, gender, ethnicity, duration and number of hemodialysis in a week and (P>0.05). Conclusion: This study, which is the first report from this area, show that the lower prevalence of anti HEV Ab in hemodialysis patients in comparison with pregnant and childbearing age women. Keywords: Hepatitis E Hemodialysis Elisa Gorgan

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Abstract Background and Objective: Resistance to antiretroviral agents is a significant concern in clinical management of HIV-infected individuals. Resistance is the result of mutations that develops in the viral protein targeted by antiretroviral agents. Material and Methods: In this cross-sectional study, the blood samples of 40 HIV-positive patients were collected. Twenty of them were drug-naïve and the rest were under treatment for at least one year by antiretroviral agents. Virus genome was extracted from patient's plasma with high-pure-viral-nucleic-acid kit. Then, by means of reverse-transcriptase and specific primers of protease genes were amplified and sequenced. Sequences of genes, drug- antiretroviral- resistant mutations and subtypes were determined using Stanford University’s HIV-drug-resistance databases. Results: Drug-naive patients show 15% resistance to nucleoside-reverse-transcriptase inhibitor (NRTI) and 20% resistance to non-nucleoside-reverse-transcriptase inhibitor (NNRTI). Anti-protease resistance is not observed in any patients. In under treatment patients, drug resistance to NNRTI (25%) is more than drug resistance to NRTI (20%) and the rate of drug resistance to protease inhibitor is 5%. Conclusion: Our findings show a high prevalence of drug-resistant mutations in Iranian-drug-naïve-HIV-infected patients. But in under treatment individuals, the rate of drug resistance is less than previous studies. Keywords: HIV Nucleoside Inhibitor Non-Nucleoside Inhibitor Protease Inhibitor

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Abstract Background and Objective: The main cause of spreading staphylococcal infections among patients is the healthy carriers working in hospitals. With the secretion of different sorts of toxins such as entrotoxin, this bacteria can provide the conditions for attacking on the host. The main objective of this study is identification of the characteristics and differences in the Staphylococcus aureus isolated from healthy carriers and from the patients on the basis of enterotoxin genes (sea-see). Material and Methods: One hundred and twenty of the patients and 80 of healthy carriers worked in health centers of Gorgan, north of Iran, were investigated for S. aureus isolate. The isolates were evaluated by PCR for Enterotoxin Genes A-E (SEA to SEE). Results: Enterotoxin genes (SEA to SEE) was found in 87.5% of the total isolates and the most frequent one was enterotoxin gene sea (N= 124). The prevalence of these isolates in healthy carriers was significantly higher than those of the patients. Conclusion: Based on the results, the high percentage of S. aureus isolated from clinical samples contains enterotoxin genes. Therefore, Human as the source and carrier of S. aureus is paramount importance, which is due to significant relationship between being toxigenic strains and the source of isolation. Key words: Staphylococcus Aureus Enterotoxin Patient Carrier

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Abstract Background and Objective: The emergence of a novel H1N1influenza A virus of animal origin with transmissibility from human to human poses pandemic concern. Current subtypes of Seasonal influenza A viruses spread in human are influenza A H1N1 influenza A H3N2 and influenza type B viruses. The aim of this study was to determine current strains of the H3N2 and new H1N1 subtypes of influenza A virus from patients suspected influenza infection in 2009 flu pandemic in Golestan province, Iran. Material and Methods: In this descriptive study, respiratory samples (n = 153) from patients with acute respiratory symptoms were collected in 2009 flu pandemic applied during 2009 pandemic influenza in Golestan province. After reverse transcription of extracted viral RNA, PCR was developed for both H1N1and H3N2subtypes using CDC specific primers. Results: The mean age of patients was 16.59. Of them 45.1% were male. Thirteen (8.49%) were infected with seasonal influenza H1N1 and 25(16.33%) with seasonal H3N2influenza. Conclusion: The rate of infection with seasonal H1N1and H3N2is similar to other studies reported from Iran, but lower than the rate reported from other parts of the world. Key Words: Influenza A Virus, H1N1, H3N2, RT-PCR, Iran

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Abstract Background and Objective: Bacterial colonization in upper respiratory airways is one of the major risk factors for the development of the ventilator–associated pneumonia (VAP), which is the most common and serious hospital-acquired infection in intensive care unit (ICU). The aim of this study was to determine the frequency of oropharyngeal microorganisms of patients with tracheal tube hospitalized in ICU. Material and Methods: Of 39 patients hospitalized in ICU of panje Azar Hospital, the oropharyngeal cultures were taken after admission. The samples were evaluated for growth of Staphylococcus aureus, Pneumococcus, Enterococcus, Pseudomonas, and E-coli. Results: The mean age of the patients (21 men, 18 women) was 43.64±15.01. The culture was positive in 28.2% and the most common isolate was Pseudomonas aeruginosa (10.3%). Conclusion: Pseudomonas, which is the main pathogen for ventilator- associated pneumonia, may be a potential threat for the patients hospitalized in intensive care units. Keywords: Microbial Colonization, Endotracheal Tube, Intensive Care Unit, Ventilator Associated Pneumonia

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Abstract Mycobacterium genus, including pathogenic and environmental species, is called non-tuberculosis mycobacteria. In this review, we assessed the research about the frequency of non-tuberculosis mycobacteria in Iran. The analyses showed that there are 16 and 28 mycobacterial species isolated in water and soil samples, respectively. The most frequent mycobacterial species in water were M. fortuitum (25.4%) and M. chelonae (25.4%), and in soil it was M. fortuitum (19.7%). The most frequent species in clinical samples was M. fortuitum, too. The frequency of non-tuberculosis mycobacteria in various clinical samples was various, and on average 1.1% of the suspected tuberculosis clients referred to the healthcare centers have non-tuberculosis mycobacteria. Keywords: Environmental Mycobacteria, Non-Tuberculous Mycobacteria, Iran, M. fortuitum

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Abstract Background and Objective: Biofilm is a complex microbial community embedded in a self-produced extracellular polymeric matrix. We aimed to study the extent of biofilm formation by S. Areas isolates and its relation to some phenotypic and genotypic criteria. Material and Methods: One hundred-fifty strains of Staphylococcus aureus isolated from Gorgan were studied. Microtiter plate assay method was used for investigation of biofilm formation.The biofilm formation of strains were recorded and its relation to accessory gene regulator (agr) and antibiotic resistance were assessed by X2 test. Results: Eighty-four isolates (56%) were able to form biofilm. The strength of biofilm formation in agr group I was more than that of other groups. The biofilm formation among S. Areas isolated from the wound and urine (both with 75 %) had the highest capability. Methicillin-resistant isolates had a greater ability to biofilm formation. Conclusion: Methicillin resistant isolates had a greater ability to biofilm formation. Given the importance and treatment related problems of Methicillin-Resistant Staphylococcus Aureus (MRSA) especially Community Acquired-Methicillin-Resistant Staphylococcus Aureus (CA-MRSA), it is a necessity to control or remove the biofilm formation alongside antibiotic treatment. Keywords: Staphylococcus Aureus, Biofilm, Microtiter Plates Assay, PCR

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Background and Objective: Cytomegalovirus (CMV), one of the most common opportunistic pathogens in patients infected with human immunodeficiency virus (HIV), can cause the diseases such as encephalitis, pneumonia, and chorioretinitis. This study aimed at molecular studying of CMV infection in individuals infected with the human immunodeficiency virus. Material and Methods: In this study, 50 venous blood samples from HIV-infected individuals were taken. Patients were divided into two categories: patients under treatment with and without antiretroviral drugs. Plasma were separated from blood samples and examined for the presence of cytomegalovirus genome by PCR. Material and Methods: this study was conducted on 50 blood samples from HIV-infected individuals, and plasma was separated and examined for the presence of cytomegalovirus genome by PCR. Patients were divided into two group of under treatment with and without antiretroviral drugs. Results: Of 50, 28 (% 56) were men and 22 (% 44) were women. CMV genome was identified in 8 samples (16%), and the molecular prevalence of CMV infection was 21.4% (n= 6) in males and 9.1% (n = 2) in females. Conclusion: Given the frequency of Cytomegalovirus Active Infection in HIV-infected individuals under antiretroviral therapy, we should be careful about the treatment of Cytomegalovirus Active Infection. Keywords: Active Infection, Cytomegalovirus, Human Immunodeficiency Virus, Shiraz, PCR

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Abstract Background and objective: The periodic evaluation of antimicrobial activity of different antibiotic is essential because antibiotic sensitivity pattern may also changed during short courses. The aim of this study was to assess the frequency of Multi-drug Resistance (MDR) in Gram negative uropathogens. Material and Methods: This study was conducted on 111 gram negative uropathogens using standard microbiology methods in Gorgan, 2011-2012. Antibiotic susceptibility was investigated by Kirby-Bauer disk diffusion methods (DDM). Results: the most common isolates were klebsiella ( 40.5%) , Enterobacter (26.1%) , pseudomonas (13.5%) , proteus( 6.3%) , acinetobacter (1.8% ) and other gram negative bacteria ( 18.3%) .The highest antibiotic resistance was seen to clindamycin (99.1%), and the most sensitivity to Carbapenems (94.6%).Multi drug resistant was seen in 68.5% of isolates. In inpatients, all of the citrobacter species had resistant to multi drugs simultaneously. Conclusion:a high frequency of multi drug resistant in uropathogens is observed in both inpatients and outpatients. Keywords: Multi Drug Resistant, Gram Negative Bacteria, Urinary Tract Infection

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Abstract Background and Objective: Human Cytomegalovirus (CMV) is an important cause of congenital viral infection that can lead to serious diseases and complications in infants. Application of rapid, sensitive, and specific HCMV detection methods is necessary for congenital infection detection. We aimed to optimize the use of PCR and ELISA for detection of HCMV in infants. Material and Methods: PCR–ELISA was performed by using specific primers and probe for detection of the HCMV glycoprotein B gene. First, the extracted DNA from urine samples and controls were labeled by digoxigenin during DIG-labeling PCR. After that, Biotin-labeled probe captured the DIG-labeled PCR products. The probe-PCR product hybrid is immobilized on a streptavidin-coated Microtiter plate, and detection was confirmed by proxidase-conjugated anti-digoxigenin antibody, and calorimetric substrate. Results: The clinical Human CMV strains isolated from16 patients were detected by this method. The optimized PCR-ELISA method was able to detect less than100 copies of HCMV genome. There was no non-specific reaction. Conclusion: PCR-ELISA can be applied as a sensitive, specific and reliable method for Semi-quantitative CMV detection in clinical samples. Keywords: Cytomegalovirus, Glycoprotein B, PCR-ELISA, Semi-Quantitative

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Abstract Background and Objective: Along with antibiotics, the use of biological methods to combat bacteria is notably considered. A natural barrier such as amniotic membrane is one of the ways of dealing with bacterial infections. The aim of this study was to determine the antibacterial effect of human amniotic membrane. Materials and Methods: This descriptive study was performed in Dezyani teaching Hospital of Gorgan University of Medical Sciences, Iran. To evaluate the antibacterial activity against Staphylococcus aureus, Pseudomonas aeruginosa and Escherichia coli bacteria, 20 amniotic membranes were obtained from postpartum mothers and examined by repeated dilution, diffusion and extraction techniques. Data were collected by observation method and described by mean and standard deviation. Results: The antibacterial activity was found in 15% of the samples against Staphylococcus Aureus and Pseudomonas aeruginosa, while no antibacterial activity was found against E. coli. Given the 15% positive responses, "Diffusion" and "repeated dilution" techniques were more effective in investigating the antibacterial effect of amniotic membrane. Conclusion: The results show the probability of antimicrobial effect of amniotic membrane tissue and it seems that this property can be affected by many factors. Keywords: Amniotic Membrane, Anti-Bacterial Properties, Laboratory Methods

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Abstract Background and Objective: Klebsiella pneumoniae is one of the agents causing nosocomial infection therefore, we decided to report the prevalence of Klebsiella pneumoniae caused infection. Material and methods: The frequency of Klebsiella in culture media samples of Panje Azar hospital was studied in 2011-2012. After determination of the species with biochemical methods and determination of resistance to third generation cephalosporins, the existence of responsible genes for this resistance was investigated using specific primers. The PCR product for CTX-M gene was sequenced. Results: During the study, 70 isolates of Klebsiella were isolated in that 51 (72.8%) related to three months of November, December and January. Except for the one related to November, other ESBL cases belonged to these three months. Based on molecular investigation of ESBL genes, these isolates at least were in 3 types and had a high frequency in Internal, female and Emergency wards. Conclusion: The present report implied a sudden prevalence of Klebsiella pneumoniae that detected and controlled by a correct monitoring. Keyword: Klebsiella Pneumoniae, ESBL, CTX-M

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Abstract Background and Objective: The M2 gene expressing the conserved protein in influenza virus can be used to make a single-dose vaccine with permanent immunity. Material and Methods: The mice were allocated to one case group immunized with pcDNA3-M2 and two control groups with pcDNA and PBS, in three dozes with interval of two weeks. Two weeks after the last injection, Cellular immunity was analyzed by MTT lymphocyte proliferation, interferon gamma (IFN-gamma) and interleukin 4 (IL-4) ratio assays. The remaining animals were challenged with PR8 virus. Results: The production rate of IFN8 and IL4 in pcDNA - M2 group was higher than that of control groups (P >0.0001). Given the results of lymphocyte proliferation, Stimulation index (SI) in vaccinated mice was significantly higher than that of control groups (P<0.05). In comparison with mortality rate of 100% in control groups , the animals Challenged with PR8 vaccine had a 50% fatal rate implying a high protection level for this vaccine. Conclusion: The pcDNA3-M2 Vaccine can be considered as a promising vaccine against influenza infections. Keywords: Influenza Virus, Gene Vaccine, M2 Protein