Medical Laboratory Journal

Prevalence of Echinococcus Granulosus in Carnivores of Iran: Systematic Review and Meta-Analysis Study

J Abdi

Abstract In this study, the prevalence of Echinococcus granulosus was surveyed by meta-analysis and systematic review. Using main keywords of Echinococcus granulosus, dog and carnivores, we searched standard sites such as Google scholar, SID, Pubmed, Iranmedex, Irandoc, Science direct and magiran. Fifteen reliable articles were reviewed and analyzed by STATA software, Ver 11. From obtained articles, the number of 2271 dogs, 167 jackals and 174 foxes have been investigated by Iranian researchers. The infection rate of E.granulosus was reported 27.4% (95% CI, 20.9-33.9) in this study in that the highest from north and north west of Iran with prevalence of 33.3 %(CI:22.1-44.4), the lowest by 21.9 %(CI:12.5-31.3) from west and south west of Iran and the average from the central region of Iran ( 26.9% CI:12.8-41). Among cities the highest (64%) and the lowest (3.3%) infection rate were reported in Garmsar city (2010) and Sistan and Balochestan province (1997). Given that the overall prevalence of E.granulosus in Iran is high, Control and prevention measures including destruction of stray dogs and treatment of owner dogs seems to be necessary. Keywords: Echinococcus Granulosus, Carnivores, Iran, Systematic Review

YMDD and FLLAQ Mutations in Chronic Hepatitis B Patients

Tabarraei, A

Abstract Background and Objective: Lamivudine is the first orally available drug approved for treatment of chronic hepatitis B. Mutations at the YMDD and FLLAQ motifs in the domains of HBV polymerase gene contribute resistance to lamivudine. This study was aimed to determine the rate of YMDD and FLLAQ mutants in hepatitis B patients in Golestan Province, Iran. Material and methods: In this cross sectional study, 120 patients with chronic HBV infection were recruited. Of them, 55 were treated and 65 untreated with Lamivudine. HBV DNA extractions from plasma and polymerase chain reaction (PCR) were performed. For detection of Lamivudine mutants direct sequencing and alignment of products were applied using reference sequence from Gene Bank database. Results: the average age of patients was 36.31±10.07, which 35% of them were female and 65% were male. Mutations at the YMDD and FLLAQ motifs in the domains of HBV polymerase gene were detected in 12 of 55 patients (21.81%) treated with Lamivudine while no mutation was observed in in untreated patients. The YMDD and FLLAQ mutants were detected in 9.16% (11/120) and 0.83% (1/120) of chronic HBV patients, respectively. Conclusion: Usual HBV mutations, which play an important role in lamivudine resistance, detected in this study are similar to other studies. Key words: Hepatitis B Viruse, YMDD Mutation, Lamivudine, Iran.

Optimization of PCR-ELISA in Detection of Human Cytomegalovirus Infection

A Tabarraei, A

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

Serotyping of Salmonella in Unpasteurized Cream Samples and Their Antibiotic Resistance Pattern

Soltan Dallal, MM

Abstract Background and Objective: Cream, a rich dairy product, with a neutral PH and low preservation time is a suitable medium for microbial growth. Salmonella is one of the most important pathogens in causing food poisoning and human gastroenteritis. This study aimed at investigating the quality of traditional cream for the bacterial contamination. Material and Methods: In this cross-sectional study, 196 non-pasteurized cream samples were collected from 5 regions of Hamedan, Iran. After dilution in phosphate buffer and serial dilution preparation, Salmonella was transferred to Rappaport-Vassiliadis (RV) enrichment medium according to CDC guidelines. After 24 h incubation at 37 ° C, a loop was inoculated in MacConkey and Hektoen Enteric (HE) Agar. The suspected colony phenotype was examined and their identification confirmed by API-20 E. Results: The samples (29%) were contaminated with at least one kind of bacteria, Salmonella Spp (4.59%) and Yersinia Spp (2.55%). The other bacteria like Escherichia, Enterobacter, Klebsiella, and Citrobacter were also isolated. Nine samples were contaminated with two kinds of bacteria. Conclusion: The presence of bacteria such as Salmonella and Yersinia in unpasteurized cream indicates that more quality control needs to be applied to the traditional crème produced in the city by health control office of food products. Keywords: Cream, Salmonella SPP, Coliform, Yersinia Enterocolitica, Hamedan

Antibacterial Effect of Human Amnion Membrane

Okhly, M

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

rBCSP31 Antibody Response in Patients with Brucellosis: A Candidate for Brucella Vaccine

A Mohabati Mobarez

Abstract Background and Objective: One of the proteins shared in all strains of Brucella is 31 kDa surface protein (BCPS31) that could be an appropriate target for immunization and serological diagnosis. Material and Methods: In the present study, BCSP31 produced as a recombinant protein in pET28a (+) expression system was utilized, using ELISA, to detect trace specific antibody (IgG) in brucellosis patients' serum that was confirmed by culture. We also evaluated cytokine response of peripheral blood lymphocytes to this protein in the cell culture. Results: The results indicated a significant amount of surface protein antibodies (IgG) in the serum of patients with brucellosis. Evaluation of lymphocyte responses to rBCSP31 also showed a significant IL-12 and IFN-γ production in patients’ lymphocyte cultures. Conclusion: These results suggest that BCSP31 can elicit specific humoral and cellular responses during host infection and it can be used in designing immunization and serologic diagnosis systems. Keywords: Brucellosis, 31kDa Cell Surface Protein, Brucella, Cytokine

Prevalence of Klebsiella Pneumoniae in Panje Azar Hospital of Gorgan, Iran, 2011

Ghaemi, EA

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

Antimicrobial Activity of an Alcaligenes faecalis Strain Isolated from Oil Contaminated Soil

Ebrahimipour, GH

Abstract Background and Objective: The bacteria living in the specific ecological conditions are among the most promising antimicrobial producers. This study aimed at isolating antimicrobial producing bacteria from soils contaminated with crude oil. Material and Methods: the samples were obtained from crude oil contaminated soils around Dezful located in Khuzestan province, Iran, and antimicrobial producing bacteria were isolated using disc diffusion and cross streak culture. Then, the best bacterium was selected and its antimicrobial potency was studied against indicator microorganisms. The isolate was also characterized based on biochemical properties and phylogenetic analysis. Results: based on the results, the highest antimicrobial activity of isolated bacterium was related to Candida albicans, Aspergillus niger, Bacillus subtilis, E. coli and Klebsiella pneumonia. An intermediate effect was determined against Serratia marcesens and Staphylococcus aureus, whereas no effect was observed against three strains of Enterococcus. Using biochemical characteristics and phenotypic traits, the isolate was identified as Alcaligenes faecalis. Conclusion: given that the isolate has broad spectrum activity against a various range of microorganisms and in comparison with some antimicrobial compounds produced by other Alcaligenes species, it seems the novelty of this antimicrobial compound. Keywords: Antimicrobial Compound, Oil Contaminated Soil, Alcaligenes faecalis

Antibiotic Susceptibility of Pseudomonas Aeruginosa Isolated from Cystic Fibrosis Patients

Ghasemi, F

Abstract Background and Objective: Cystic fibrosis (CF) is an autosomal recessive genetic disease and Pseudomonas aeruginosa is one of the most common bacteria colonized in CF patients. Growing resistance of this bacterium to antibiotics now a day is a challenge of controlling infection in CF patient. In this study colonization of CF patients with Pseudomonas aeruginosa and antibiotic susceptibility pattern of isolated strains were examined. Material and Methods: From 100 CF patients, during a year, sputum and bronchial swabs were collected. After culturing the samples, some of them were reported as Pseudomonas aeruginosa using biochemical tests. Mucoid strains of Pseudomonas aeruginosa were identified the same as non-producing alginate strains while for catching single pure colony, repeated passage was used. For determining antibiotic resistance of Pseudomonas aeruginosa to some antimicrobial agents Kirby-Bauer method based on CLSI was used. Results: Of 100 samples, 40 (40%) were positive for Pseudompnas aeruginosa. The prevalence of P. aeruginosa was 23.8, 36.84 and 80% at the age of 1-3, 4-12 and 13, respectively. Conclusion: Statistically, there is a significant difference between age and contracting with Pseudomonas aeruginosa in that the higher the age the more colonization with Pseudomonas aeruginosa. Key words: Pseudomonas Aeruginosa, Cystic Fibrosis, Drug Resistance

Prevalance of vanA Gene in Vancomycine Resistant Staphylococcus Aureus (VRSA) Isolated from Chicken and Turkey Meat in Tehran, Iran

Hosseini Doust, SA

Abstract Background and Objective: Staphylococcus aureus have been isolated frequently from the infection of blood, skin and soft tissue, and is one of the causative agents of food-borne illnesses. The purpose of this study was to investigate the occurrence of vanA in raw meat samples in Tehran, Iran. Material and Methods: Totally, 119 samples of chicken and turkey raw meat were cultured for Staphylococcus aureus. The Resistance to vancomycin was determined and Van A Gene detected by PCR method. Results: twenty- nine strains of Staphylococcus aureus were isolated. Of these, 14 (48.5%) were fully resistant to vancomycin and six of them had van A gene. Conclusion: Given that a large number of the samples were infected to VRSA and diversity of quality in production and distribution of foods and meat, the results could be varied. To avoid the transmission of antibiotic resistance chain to human, we should pay attention to make regulation and health conditions in production and distribution. Keywords: Staphylococcus Aureus, Vancomycin Resistance, Van A, Turkey Raw Meat, Chicken

Glycated Hemoglobin, Gastric Juice Nitric Oxide and Oxidative Stress in Diabetic Patients Infected by Helicobacter Pylori

H Dolatkhah

Abstract Background and Objective: Recently, diabetes mellitus has been known as one of the main cause of upper gastrointestinal symptoms. Since a high prevalence of H. Pylori in diabetic patients has been reported, we aimed to evaluate the level of gastric juice Nitric Oxide (NO°), Oxidative Stress and Glycated Hemoglobin. Material and Methods: In case group, the participants were 60 diabetic patients infected with H. Pylori, and in control groups 60 diabetic patients without H. Pylori and 60 healthy individuals. The level of NO° in gastric juice was measured calorimetrically and the activity of superoxide dismutase (SOD) and glutathione peroxidase (GPX) in gastric biopsy was determined using standard methods. The percentage of Glycated Hemoglobin (HbA1C) was measured by ion exchange chromatography. Results: In case group compared to controls, significantly increased level of blood HbA1C, nitric oxide in gastric juice, activity of SOD and GPX in the gastric mucosa were observed (p<0.0001). Conclusion: A significant increase of glycated hemoglobin in diabetic patients with H. Pylori and high activity of antioxidant enzymes in the case group may indicate a high production of reactive oxygen species and the presence of oxidative stress in these patients. Key Words: Diabetes Mellitus, H. Pylori Infection, Glycated Hb, Nitric Oxide, Oxidative Stress

Relationship between Ferritin Serum Level and Attention Deficit Hyperactivity Disorder

MT Badele

Abstract Background and Objective: With regard to high prevalence of attention deficit hyperactivity disorder (ADHD) and its being significantly affected by nutritional factors, we aimed to determine the relationship between Ferritin serum level and ADHD. Material and methods: This ex-post- facto (causal comparative research) design study was conducted on 60 children, selected via convenience sampling. Thirty of them were ADHD children diagnosed by a psychiatrist using DSM IV checklist, as a case group, and the rest were healthy ones located in control group. Having their family informed consent, their Ferritin level was measured via ELIZA method. Results: The results show that Ferritin serum level of ADHD children are lower than that of healthy ones. Using t- test, it was indicated that the difference is significant (p= 0.002). Besides, the result of Pearson correlation coefficient showed that there is no significant relation between Ferritin and ADHD. Conclusion: In terms of the results and the importance of this issue, we recommend conducting further controlled research. Keywords: Ferritin, ADHD, Attention Deficit Hyperactivity Disorder

Identification of Malassezia Species Isolated from Patients with Seborrheic Dermatitis Using PCR-RFLP Method in Arak, Iran

Didehdar, M

Abstract Background and Objective: Malassezia that is a part of normal flora is lipophilic yeast involved in a variety of skin diseases such as seborrheic dermatitis, pityriasis versicolor, atopic dermatitis and psoriasis. Seborrheic dermatitis affects most often the sebaceous-gland-rich areas of skin such as face, scalp, and parts of the upper trunk. Dandruff is a mild variant of seborrheic dermatitis characterized by scaling. In this study, Malassezia species causing dandruff were identified. Material and Methods: In this descriptive study, the samples (n= 60) from participants with dandruff were examined under a microscope using 10% KOH solution and cultured in Leeming and Notman ager medium. DNA Extraction was performed from colonies by glass bead and the Malassezia genus, and species were detected by CfoI enzyme using PCR-RFLP method Results: Of 60, 40 (66.6%) were positive for Malassezia yeast. The positive samples in direct examination grew in culture medium. Malassezia species isolated were Malassezia globosa (25 cases), Malassezia restricta (10 cases), Malassezia furfur (3 cases) and Malassezia sympodialis (2 cases). Conclusions: In most studies, the Malassezia species were identified as the agents causing seborrheic dermatitis. In our study, Malassezia globosa was isolated as a dominant species. Keywords: Seborrheic Dermatitis, Malassezia SPP, Arak

Lipase Gene Expression of Resistant and Sensitive Candida Albicans to Fluconazole Isolated from Patients Suffering from Oral Candidiasis and Vaginal Candidiasis

Nasrollahi Omran, A

Abstract Background and Objective: With the development of drug resistance in strains of fungi, there is a considerable resistance of Candida albicans strains to fluconazole. Molecular studies are developing to determine the relationship of such a drug resistance with the increased gene expression of enzymes produced in drug-resistant Candida isolates. We aimed to evaluate the relationship between extracellular lipase gene (LIP8) expression of Candida albicans isolated from candidiasis and sensitivity or resistance to fluconazole. Material and Methods: Drug susceptibility of Candida albicans was performed in oral and vaginal candidiasis to determine the proportion of strains sensitive or resistant to fluconazole using NCCLS method. To evaluate and compare the expression of these genes in the susceptible and resistant strains, RT real-time PCR reaction was used. Results: Of 46 Candida albicans, 20 were susceptible, 12 were semi-susceptible and 14 were resistant to fluconazole. By using PCR reaction, the results showed that the expression of this gene in fluconazole-susceptible isolates was moderate, while it was high in the isolates resistant to fluconazole. Conclusion: The results of lipase gene (LIP8) expression showed that the additional expression of some genes of the enzymes responsible for virulence of Candida may also play a role in resistance to fluconazole. Keywords: Candidiasis, Lipase Gene Expression, RT real-time PCR, Fluconazole

Comparison of Routine Method with Antibody and Antigen Ones for Diagnosing Giardia-Entamoeba Histolytica in Stool and Blood

Roozbehani, M

Abstract Background and Objectives: Giardia lamblia and Entamoeba histolytica are the most prevalent human intestinal pathogenic protozoa, worldwide. The clinical features of Giardia infection are acute diarrhea, a chronic condition with continuous diarrhea and malabsorption. Entamoeba histolytica invade intestinal tract without any typical clinical indications, and it can involve liver and other organs too. Therefore, we aimed to study these protozoa by serological and parasitological methods. Material and Methods: In this comparative study, the stool and blood specimens were collected from 1025 patients selected via simple random sampling in three different laboratories located in Tehran and Karaj, Iran (2012). Formalin Detergent test was performed on all samples. Both serum and stool positive samples of this method were analyzed for antigen and antibodies related to Giardia lamblia and Entamoeba histolytica, respectively. Results: of 1025 stool specimens, 76 (4.7%) were positive for Giardia lamblia and 19 (1.8%) for Entamoeba histolytica using Formalin-detergent method. In ELISA, 81 (7.9%) coproantibodies to Giardia lamblia and 24 (2.3%) coproantibodies to Entamoeba histolytica, 78 (7.6%) corproantigen for Giardia lamblia, and 5 (0.4%) for Entamoeba histolytica were observed. circulatory antibodies to Entamoeba histlytica were detected in 22 cases (2.1%) Conclusion: Sensitivity of microscopic method compared to serological methods is higher than 90% therefore, Formalin-detergent method can be the best method for stool examination. Key words: Giardia Lamblia and Entamoeba Histolytica, Coproantibody, Coproantigen, Blood Antibody

Cloning of Minor Autolysin of Streptococcus Pneumoniae

Pourmand, MR

Abstract Background and Objective: Increased antibiotic resistant strains and inadequacy of current vaccines against pneumococcal infections necessitate the study of novel protein antigens. It seems that minor autolysin of Streptococcus pneumoniae may have antigenicity. Thus, we aimed at cloning its gene for the first time. Material and Methods: After DNA extraction of Streptococcus pneumoniae (ATCC 49619), Specific primers were designed for amplifying minor autolysin gene fragment, using PCR. The purified gene fragment was inserted into pET21a vector and was transformed into bacterial competent cells by heat shock technique. The presence of gene and absence of mutation in the recombinant vector were checked out with sequencing and enzymatic digestion methods. The gene sequence was finally analyzed by bioinformatic tools. Results: The gene of minor autolysin was cloned successfully and the result of enzymatic digestion was the indication of complete isolation of this gen from plasmid. . Bioinformatics studies revealed that the mature protein was lacking signal peptide and the gene encoded 318 amino acids with a molecular weight of 36.4 kDa. Conclusion: The presentation and characterization of novel antigens such as minor autolysin could help us with finding new approaches for preventing and controlling pneumococcal infection. Keywords: Streptococcus Pneumoniae, Minor Autolysin, Cloning

Prevalence of Cryptosporidium among Diarrheic Children Hospitalized in Gonbad Kavus City, 2011

Sharbatkhori, M

Abstract Background and Objective: Cryptosporidium is a common protozoan causing diarrhea in human, specifically in children. Hence, we aimed to investigate the prevalence of this protozoan among diarrheic children hospitalized in Gonbad Kavus in 2011. Material and Methods: Three stool samples were collected from diarrheic children in two hospitals of Gonbad city and a relevant questionnaire was filled out for each child. The stool samples were concentrated by formalin ether method, and the infection was assessed by modified acid-fast staining method. Results: Of 547 children, 27 (4.9%) were infected with cryptosporidiosis. There was no significant relationship between the amount of infection and gender and habitation area (urban/ rural). The infection rate was significantly prevalent in 2-4-year-old children (P=0.013). The most and the least infection rate were observed in spring and winter, respectively (P< 0.0001). There was a significant association between the disease and keeping animal (P= 0.041) Conclusion: The prevalence of cryptosporidiosis in diarrheic children in Gonbad is almost equal to other regions of the country and keeping animal and spring season may be considered as the risk factors for the disease. Keywords: Cryptosporidium, Cryptosporidiosis, Diarrhea, Children, Golestan, Iran

Prevalence of E. coli O157: H7 Isolated from Patients with Acute Gastroenteritis

Zolfaghari, MR

Abstract Background and Objective: Diarrheal diseases may occur in all age groups, worldwide. Escherichia coli (E. coli O157: H7) is one of the most important bacterial agents causing this disease. The purpose of this study was to assess the prevalence of diarrhea in patients with acute diarrhea caused by this bacterium. Material and Methods: In this study, 214 stool samples were collected from acute diarrheic patients in hospitals and clinical laboratories of Malayer city, Iran. The samples were cultured in rich, differentiation, specific and selective medium. To detect E. coli O157: H7, rapid detection method and antisera were used . Results: In 12.15% of the samples, lactose fermentation was observed and E. coli was afirmed. The results of lactose positive samples showed that 6.54% were negative-β- Glucuronidases and were confirmed as E. coli O157. Finally, by using O157: H7 antisera, 1.87% of 14 samples that were negative- β-Glucuronidases were asserted as E. coli O157. Conclusion: based on the results, the prevalence of E. coli O157: H7 is higher in children and elders than the others. Food and water play a significant role in transferring E. coli. Keywords: Prevalence, E. coli O157: H7, Gastroenteritis, Patients