Comparative evaluation of real-time PCR and ELISA for the detection of human fascioliasis.

Fascioliasis is a zoonotic parasitic infection caused by Fasciola species in humans and animals. Despite significant advances in vaccination and new therapeutic agents, little attention has been paid to validating methods for the diagnosis of fascioliasis in humans. Serological techniques are convenient assays that significantly improves the diagnosis of Fasciola infection. However, a more sensitive method is required. The aim of this study was to compare the Real-Time PCR technique with the indirect-ELISA for the detection of Fasciola hepatica in human. Using a panel of sera from patients infected with Fasciola hepatica (n = 51), other parasitic infections (n = 7), and uninfected controls (n = 12), we optimized an ELISA which employs an excretory-secretory antigens from F. hepatica for the detection of human fascioliasis. After DNA extraction from the samples, molecular analysis was done using Real-Time PCR technique based on the Fasciola ribosomal ITS1 sequence. Of 70 patient serum samples, 44 (62.86%) samples were identified as positive F. hepatica infection using ELISA and Real-Time PCR assays. There was no cross-reaction with other parasitic diseases such as toxoplasmosis, leishmaniasis, taeniasis, hydatidosis, trichinosis, toxocariasis, and strongyloidiasis. The significant difference between the agreement and similarity of the results of patients with indirect ELISA and Real-Time PCR was 94.4% and 99.2%, respectively (Cohen's kappa ≥ 0.7; P = 0.02). Based on the Kappa agreement findings, the significant agreement between the results of ELISA and Real-Time PCR indicates the accuracy and reliability of these tests in the diagnosis of F. hepatica in humans.

Association between colorectal cancer, the frequency of Bacteroides fragilis, and the level of mismatch repair genes expression in the biopsy samples of Iranian patients.

BACKGROUND: Deficient DNA mismatch repair (MMR) can cause microsatellite instability (MSI) and is more common in colorectal cancer (CRC) patients. Understanding the carcinogenic mechanism of bacteria and their impact on cancer cells is crucial. Bacteroides fragilis (B. fragilis) has been identified as a potential promoter of tumorigenesis through the alteration of signaling pathways. This study aims to assess the expression levels of msh2, msh6, mlh1, and the relative frequency of B. fragilis in biopsy samples from CRC patients. MATERIALS AND METHODS: Based on the sequence of mlh1, msh2, and msh6 genes, B. fragilis specific 16srRNA and bacterial universal 16srRNA specific primers were selected, and the expression levels of the target genes were analyzed using the Real-Time PCR method. RESULTS: Significant increases in the expression levels of mlh1, msh2, and msh6 genes were observed in the cancer group. Additionally, the expression of these MMR genes showed a significant elevation in samples positive for B. fragilis presence. The relative frequency of B. fragilis in the cancer group demonstrated a significant rise compared to the control group. CONCLUSION: The findings suggest a potential correlation between the abundance of B. fragilis and alterations in the expression of MMR genes. Since these genes can play a role in modifying colon cancer, investigating microbial characteristics and gene expression changes in CRC could offer a viable solution for CRC diagnosis.

Characterization of class 1 integrons in metallo-ß-lactamase-producing Acinetobacter baumannii isolates from hospital environment.

BACKGROUND AND OBJECTIVE: The emergence and widespread dissemination of antibiotic resistance in A. baumannii, has become a globally challenge. The increasing hospital outbreaks by multi-drug resistant (MDR) A. baumannii strains, shows the necessity of continuous monitoring to find sources of resistant strains in hospitals. This study aimed to identify the presence of class 1 integrons and metallo-ß-lactamase (MBL) related genes in A. baumannii isolates from hospital environment. METHODS: In order to identify A. baumannii isolates, a total of 297 environmental samples were collected from burn wards and intensive care units (ICUs) of two university hospitals. Resistance to common antibiotics was studied by disk diffusion method and microbroth dilution assay was used to determine the minimum inhibitory concentrations (MICs) of imipenem, colistin and tigecycline. The A. baumannii isolates were studied by polymerase chain reaction (PCR) for the presence of class 1 integrons (intI1, intl CS) and metallo-ß-lactamases (MBLs) (blaIMP, blaVIM, blaNDM) genes. RESULTS: A. baumannii was identified in 68/297 (22.9%) of hospital environment. All A. baumannii strains were multidrug-resistant (MDR), but none of them were resistant to colistin, tigecycline and ampicillin-sulbactam. All (100%) and 38 (95.0%) of A. baumannii isolates from ICUs and burn wards were imipenem resistant respectively. Class 1 integrons was identified in 30/40 (75.0%) and 23/28 (82.1%) isolates from burn wards and ICUs respectively. Two different types of gene cassettes were identified, which included: arr-2, ereC, aadA1, cmlA5 and arr2, cmlA5. MBL genes including blaVIM and blaIMP were detected in 26/28 (92.8%), 27/28(96.4%) and 39/40 (97.5%) and 31/40 (77.5%) of the isolates from the ICUs and the burn wards respectively. None of the isolates contained the blaNDM-1 gene. CONCLUSION: The findings of the present study showed that the isolation rate of MBL producing carbapenem-resistant A. baumannii (CRAB) was relatively high in the environmental surface of burn wards and ICUs, which can be considered as a potential source of outbreaks in hospitalized patients.

Molecular investigation of Toxocara infection from the serum of people living with HIV in Alborz, Iran.

BACKGROUND: Toxocara infection is one of the most common neglected infections of poverty and a helminthiasis of global importance. Traditional diagnostic methods such as antibodies detection in serum samples are limited due to cross-reactivity and poor sensitivity. The use of molecular base methods for diagnosis of Toxocara infection in Iran has not been fully explored. The purpose of the current study was to estimate the prevalence of Toxocara infection from serum samples of people living with HIV in Alborz province, Iran using serological and molecular methods. METHODS: Blood samples were collected from 105 people living with HIV. Epidemiological data of participant were obtained through a structured questionnaire to investigate the risk factors. Patients CD4+ T cell count were recorded. Anti-Toxocara IgG antibodies were detected by ELISA, with a cut-off point of 11. PCR was performed to detect genetic material of Toxocara species in the serum samples. RESULTS: The mean CD4+ count in HIV-infected individuals with positive toxocariasis serology was 255.1 ± 21.6 cells/µL. Seropositivity for Toxocara species was observed in 12/105 (11.4%) people living with HIV. Three samples gave positive results on PCR analysis. Based on the data, a statistically significant relationship was found between anti-Toxocara IgG antibodies seropositivity and underlying conditions (p = 0.017). No significant statistical association was observed between seropositivity for Toxocara and gender, age, exposure to domestic animals or pet keeping, education levels, and occupation (p > 0.05). The findings of PCR confirmed Toxocara DNA in 3/12 (25.0%) serum samples. CONCLUSION: These findings demonstrated for the first time that people living with HIV from Alborz province, are being exposed to this zoonosis and a relatively high seroprevalence of Toxocara in HIV/AIDS people needs comprehensive health education regarding personal hygiene and how to avoid exposure to this parasite infection, especially in people with an impaired immune system.

Clonal relatedness of carbapenem-resistant Acinetobacter baumannii: high prevalence of ST136pas in a burn center.

BACKGROUND: Carbapenem-resistant Acinetobacter baumannii (CRAB) is a global health crisis. This study aimed to determine the clonal relatedness of antibiotic-resistant A. baumannii isolates in hospitalized patients who suffered from burn wound infection. METHODS: One hundred and six A. baumannii isolates from 562 patients with burn wound infections, were identified and examined for antimicrobial susceptibility. Detection and characterization of carbapenem-hydrolyzing class D OXA-type beta-lactamases (CHDLs) were performed by PCR assays. The clonal relatedness of A. baumannii isolates was determined by multilocus sequence typing (MLST) according to the Pasteur scheme, dual-sequence typing of blaOXA-51-like and ampC genes, and RAPD-PCR method. RESULTS: All isolates were carbapenem-resistant while susceptible to colistin, minocycline, doxycycline, and ampicillin-sulbactam. The intrinsic blaOXA-51-like was detected in all isolates, and blaOXA-23-like was identified in 92.5% of isolates. However, blaOXA-143-like and blaOXA-58-like genes were not detected among isolates. Four distinct blaOXA-51-like alleles were determined as follows: blaOXA-317 (67.0%), blaOXA-90 (9.4%), blaOXA-69 (17.0%), and blaOXA-64 (6.6%) and four ampC (blaADC) allele types including ampC-25 (6.6%), ampC-39 (9.4%), ampC-1 (17.0%), and blaADC-88 (67.0%) were identified. MLST (Pasteur scheme) analysis revealed four ST types including ST136 (singleton), ST1 (CC1), ST25 (CC25), and ST78 (singleton) in 71, 18, 7, and 10 of A. baumannii strains, respectively. Five RAPD clusters including A (1.9%), B (26.4%), C (57.5%), D (7.5%), and E (1.9%) were characterized and 5 (4.7%) strains were found to be singletons. CONCLUSION: The present study demonstrated that there was a high prevalence of blaOXA-23-like producing CRAB in the clinical setting. The majority of isolates belonged to ST136 (singleton). However, blaOXA-23-like producing multi-drug resistant international clones including ST1, and emerging lineages (e.g. ST25 and ST78) were also identified. Interestingly, in this study ST2 was not detected.

Detection of multidrug-resistant Acinetobacter baumannii from burn patients and healthcare workers in Iran.

Multidrug-resistant (MDR) Acinetobacter baumannii is a serious global health threat. Burn patients are at high risk to acquire A. baumannii infections from endogenous sources. This study evaluated carbapenem resistance and clonal relatedness of A. baumannii isolated from burn patients and healthcare workers (HCWs).The study was performed in 100 non-duplicated A. baumannii isolates from nasal and hand samples of hospitalized burn patients and HCWs in two hospitals of Iran from June 2020 to August 2021. Antimicrobial susceptibility testing was performed and carbapenemase genes were detected by PCR. Clonal relatedness of A. baumannii isolates was determined by two single-locus sequence-based typing of blaOXA-51-like and ampC and by multilocus sequence typing (MLST).All A. baumannii isolates were found to be MDR while susceptible to colistin. The intI1, conserved segments of class 1 integron (intI1 CS), blaIMP, blaVIM, blaOXA-51-like, and blaOXA-23-like, genes were detected in 32.5%, 29.1%, 36%, 95.3%, 100%, 100%; and 14.3%, 14.3%, 21.4%, 92.9%, 100%, and 85.7% of isolates from patients and from healthcare workers, respectively. The blaOXA-58, and blaOXA-143 were not detected among the isolates. Using dual-locus blaOXA-51-like and ampC sequence-based typing (SBT), the isolates obtained from nasal samples of burn patients were grouped into 3 clusters including blaOXA-317, blaADC-88 (72.1%); blaOXA-64, ampC-25 (18.6%); and blaOXA-69, ampC-1 (9.3%). While only allele type blaOXA-317, blaADC-88 was determined among isolates from HCWs. MLST results showed A. baumannii ST136, ST25, and ST1 from burn patients. However, A. baumannii strains from HCWs belonged to ST136. Our findings indicate high prevalence of globally spreading of MDR A. baumannii ST136 carrying blaOXA-23-like from nasal and hand samples of burn patients and HCWs.

Intestinal parasites and diabetes: A systematic review and meta-analysis.

Background: Investigating the association between infectious agents and non-communicable diseases is an interesting emerging field of research. Intestinal parasites (IPs) are one of the causes of gastrointestinal complications, malnutrition, growth retardation and disturbances in host metabolism, which can play a potential role in metabolic diseases such as diabetes. The aim of the present study was to investigate the prevalence of IPs in diabetic patients and the association between IPs and diabetes. Methods: A systematic literature search was conducted from January 2000 to November 2022in published records by using PubMed, Scopus, and Web of Science databases as well as Google scholar search engine; Out of a total of 29 included studies, fourteen cross-sectional studies (2676 diabetic subjects) and 15 case-control studies (5478 diabetic/non-diabetic subjects) were reviewed. The pooled prevalence of IPs in diabetics and the Odds Ratio (OR) were evaluated by CMA V2. Results: In the current systematic review and meta-analysis, the pooled prevalence of IPs in diabetic patients was 26.5% (95% CI: 21.8-31.7%) with heterogeneity of I2 = 93.24%; P < 0.001. The highest prevalence based on geographical area was in Region of the Americas (13.3% (95% CI: 9.6-18.0)).There was significant association between the prevalence of intestinal parasites in diabetic cases compared to controls (OR, 1.72; 95% CI: 1.06-2.78). Conclusion: In line with the high prevalence of IPs in diabetic patients, significant association was found however, due to the limitations of the study, more studies should be conducted in developing countries and, the prevalence of IPs in diabetics should not be neglected.

Ocular Toxocariasis Associated with Blurred Vision and Visual Impairment: Report of Four Cases.

Ocular toxocariasis in humans is caused by infection with larvae of Toxocara species, which are common ascarid roundworms of mammals, kept in close proximity to human. Four cases with a history of contact with dogs and cats and blurred vision and visual impairment over periods of variable duration were examined. We screened patients diagnosed with ocular larva migrans syndrome between March and June 2021 at the Ophthalmology clinics affiliated with Alborz University of Medical Sciences, Karaj, Iran. Detailed demographics, clinical characteristics, and fundus photography were recorded. Anti-Toxocara antibodies in the sera and vitreous fluid detected by ELIZA. Complete recovery in all four patients was achieved following treatment with oral albendazole. The diagnosis of ocular toxocariasis can be challenging, because both the condition is relatively uncommon and its presentation varies from patient to patient. There are lots of differential diagnoses like retinoblastoma, therefore correct, quick diagnosis, and treatment is very important.

Reverse vaccinology approach to identify novel and immunogenic targets against Porphyromonas gingivalis: An in silico study.

Porphyromonas gingivalis is a primary causative agent of chronic periodontitis. Moreover, it leads to several systemic diseases, including rheumatoid arthritis, cardiovascular, neurodegenerative, and Alzheimer's diseases. It seems that the development of a vaccine against this bacterium is necessary. Thus, this study decided to identify novel immunogenic targets and developed multiple epitope-based vaccines against P. gingivalis. For this purpose, the pan/core-proteome of this bacterium was studied, and the suitable vaccine targets were selected based on different properties, including exposed localization of proteins, antigenicity, non-allergenicity, non-similarity to host proteome, stability, B-cell epitopes and MHC II binding sites, sequence conservation, molecular docking, and immune simulation. Through the quartile scoring method, 12 proteins with ≥ 20 scores were considered as suitable immunogenic targets. The results of the protein domain and functional class search showed that most of the immunogenic proteins were involved in the transport and metabolism of inorganic ions and lipids. In addition, two unknown function proteins, including WP_004584259.1 and WP_099780539.1 were detected as immunogenic targets. Three constructions carrying multi-epitopes were generated including Naked, LCL, and as chimeric structures. Among them, FliC chimeric protein had the strongest affinity to the human TLR2, 4, and 6, while the LCL platform represented the highest level of immune stimulation response. The obtained results from this study revealed new insights into prophylactic routes against P. gingivalis by introducing novel immunogenic targets. However, further investigations, including site-directed mutation and immunoassay are needed to confirm the pathogenic role and protectivity of these novel proteins.

Larva migrans in BALB/c mice experimentally infected with Toxocara cati ensured by PCR assay.

BACKGROUND: Toxocara cati, the cat roundworm, is a parasitic nematode that known to cause toxocariasis in intermediate hosts and humans. In this study, we characterized the dynamics of T. cati larvae migration in BALB/c mice after inoculation with eggs and ensured the migration detecting the larval DNA by a PCR. To evaluate the dynamics of larval migration and distribution, twenty-four BALB/c mice were orally inoculated with 2500 T. cati infective eggs and the visceral organs of the infected animals were examined by pepsin digestion and microscopic parasite counts, followed by PCR at day 1 to 28 post-inoculation. RESULTS: The PCR assays were successfully used for detection of T. cati larvae in tissue samples and T. cati larvae and the DNAs were found in the liver, lungs, heart, kidneys and the brain. We detected T. cati in 92.2% of tissue samples by PCR, 30% higher than the conventional pepsin digestion technique. CONCLUSION: Our findings demonstrated that the PCR assay is a sensitive and specific for the detection of T. cati larvae. Therefore, it could become a useful tool for the investigation of the dynamics of larval migration and Toxocara infection in murine model.

Serosurvey of anti-Toxocara antibodies and associated risk factors in domestic dogs and cats owners in Karaj, Alborz Province of Iran.

INTRODUCTION AND OBJECTIVE: Toxocariasis is a zoonotic parasitic infection with important public health considerations. The aim of the study was to assess the prevalence of anti-Toxocara species antibodies and associated risk factors in domestic dogs and cats referred by their owners to veterinary clinics located in Karaj, Alborz Province, Iran. MATERIAL AND METHODS: A cross-sectional study involving 540 owners of dogs and cats was conducted between July - December 2020. A questionnaire administered by direct interviews was used to collect socio-demographic information and data on associated risk factors. Blood samples were collected and tested by indirect enzyme-linked immunosorbent assay (ELISA). RESULTS: The overall sero-prevalence of toxocariasis among the 540 participants was 16.7% (90 of 540). When participants included in the sample were classified by age, those aged 10-29 years demonstrated higher Toxocara infection prevalence than other groups (45.6%, 41 of 90). Univariate analysis revealed that the pet owners who had contact with soil [adjusted odds ratio (AOR) = 7.61, 95% CI: 6.06-9.24, P = 0.028], practiced handwashing after contact with dogs and cats (AOR = 2.42, 95% CI: 1.15-4.85, P = 0.046), and feeding the pets with raw meat (AOR = 11.01, 95% CI: 5.21-19.43, P = 0.023) had an increased risk of acquiring toxocariasis. The study showed that demographic characteristics such as age, gender, place of residence, education, and pet's habitats were not significantly associated with toxocariasis. CONCLUSIONS: Given the findings and the progressive impact of toxocariasis in public health and its high prevalence in developing countries, including Iran, measures should be taken to inform the public about zoonoses and eliminate their putative transmission.

High prevalence of clonally related multiple resistant Salmonella Infantis carrying class 1 integrons in broiler farms.

The poultry industry in Iran is the main supplier of protein in the food chain. In the present study, we showed the importance of the possible dissemination of clonally related multiple drug resistant (MDR) Salmonella Infantis in broiler farms in Iran. In total, 156 fecal samples belonging to 23 poultry farms in Razavi Khorasan province, northeast of Iran, were examined for the presence of Salmonella serovars. Molecular serotypes and serogroups, class 1 and 2 integron types, colistin resistance genes ( mcr1 and mcr2) and antimicrobial susceptibility patterns were determined on the recovered Salmonella isolates. Based on PCR analysis, 30 recovered Salmonella isolates were identified asS. Infantis (23 isolates; 76.6%),S. Enteritidis (six isolates; 20%), and one isolate (3.3%) was not serotyped by the applied method. Class 1 integrons were detected in 22 isolates (95.6%) and class 2 integrons were not detected in any of the isolates. Although colistin resistance was prevalent in disc diffusion test, mcr-1 and mcr-2 genes were not detected. All class 1 integrons carried the cassette aadA1 gene. All Salmonella isolates were resistant to colistin and amoxicillin/clavulanic acid and MDR patterns were observed in most (96.6%) isolates. This study revealed a high prevalence rate of S. Infantis and the presence of class 1 integrons in broiler farms. The presence of the same integron cassettes in the sequenced isolates suggests that strains are clonally related. Stringent monitoring programs are required to prevent the spreading of MDR Salmonella serovars into food chain via poultry products.

Molecular study of metallo-ß-lactamases and integrons in Acinetobacter baumannii isolates from burn patients.

BACKGROUND: Productions of metallo-ß-lactamases enzymes are the most common mechanism of antibiotic resistance to all beta-lactam classes (except monobactams) in Acinetobacter baumannii. MBLs are usually associated with gene cassettes of integrons and spread easily among bacteria. The current study was performed to detect the genes encoding MBLs and integron structures in A. baumannii isolates from burn patients. METHODS: This study was performed on 106 non-duplicate A. baumannii isolates from burn patients referred to Shahid Motahari Hospital in Tehran. Antibiotic susceptibility of A. baumannii isolates was performed using disk diffusion and broth microdilution method in accordance with the CLSI guidelines. The presence of class 1 integron and associated gene cassettes as well as MBLs-encoding genes including blaVIM, and blaIMP were investigated using PCR and sequencing techniques. RESULTS: In this cross-sectional study all (100%) of the A. baumannii isolates examined were multidrug resistant. All isolates were sensitive to colistin and simultaneously all were resistant to imipenem. PCR assays showed the presence of blaVIM and blaIMP genes in 102 (96.2%) and 62 (58.5%) isolates of A. baumannii respectively. In addition, 62 (58.5%) of the A. baumannii isolates carried integron class 1, of which 49 (79.0%) were identified with at least one gene cassette. Three types of integron class 1 gene cassettes were identified including: arr2, cmlA5, qacE1 (2300 bp); arr-2, ereC, aadA1, cmlA7, qacE1 (4800 bp); and aac(3)-Ic, cmlA5 (2250 bp). CONCLUSION: A high prevalence of MBLs genes, especially blaVIM, was identified in the studied MDR A. baumannii isolates. In addition, most of the strains carried class 1 integrons. Furthermore, the gene cassettes arrays of integrons including cmlA5 and cmlA7 were detected, for the first time, in A. baumannii strains in Iran.

The combination of CipA and PBP-7/8 proteins contribute to the survival of C57BL/6 mice from sepsis of Acinetobacter baumannii.

Due to the emergence of multi-drug resistant Acinetobacter baumannii strains, there is an urgent need to develop several new strategies to control this bacterium. In this context, vaccination may be the best approach to reduce the morbidity and mortality associated with MDR isolates in vulnerable groups. Serum resistance factors have a key role in the pathogenesis of A. baumannii and can be considered as potential vaccine candidates. This project aimed to evaluate the immunological reactivity of CipA and PBP-7/8 as two serum resistance factors in a combination form against sepsis infections of A. baumannii. Recombinant proteins were obtained and immunological evaluations were performed against sepsis infection in the C57BL/6 mouse model. The data showed a statistically significant increase in total IgG levels in all three immunization regimens (CipA, PBP-7/8, and CipA + PBP-7/8) compared to the control group. The ratios of IgG2c/IgG1 in the CipA, PBP-7/8, and CipA + PBP-7/8 schedules were 8.7, 46.50, and 33.29, respectively. It appears that the immunization schedules developed a strong polarized Th1 response. The cytokine profiles of the three plans showed that IFN-γ was highly concentrated in the combination plan. However, the highest concentration of IL-17 belonged to the PBP-7/8 plan. In conclusion, the data of total IgG, survival rates and splenic bacterial loads showed that the CipA + PBP-7/8 plan was more effective than each protein individually.

Molecular analysis of methicillin-resistant Staphylococcus aureus isolates from four teaching hospitals in Iran: the emergence of novel MRSA clones.

BACKGROUND: The global spread of methicillin-resistant Staphylococcus aureus (MRSA) infections necessitates the use of validated methods for the identification and typing of this bacterium. This study aimed to determine the distribution of main molecular types of MRSA strain circulating among hospitalized patients in teaching hospitals in Isfahan and Kashan. METHODS: A total of 146 Staphylococcus aureus strains were isolated from patients in four teaching hospitals in Isfahan and Kashan during June 2017 to September 2018. The antimicrobial resistance patterns of Staphylococcus aureus strains were performed by disc diffusion method. The MRSA strains were identified phenotypically and confirmed by PCR assay. The prevalence of microbial surface components recognizing adhesive matrix molecules (MSCRAMMs) genes among MRSA strains was evaluated by multiplex PCR. The genotypes of MRSA strains were determined by multilocus sequence typing and SCCmec typing. RESULTS: Of 146 Staphylococcus aureus isolates, 24 (16.4%) isolates were identified as MRSA strains. According to antimicrobial susceptibility testing the highest resistance rates were seen for tetracycline, erythromycin, ciprofloxacin and gentamicin. All of Staphylococcus aureus isolates were susceptible to vancomycin whereas 3 (2.1%) isolates were resistant to linezolid. Three different SCCmec types were obtained among MRSA strains including 16 (66.7%) SCCmec type V, 3 (12.5%) SCCmec type III and 5 (20.8%) SCCmec type II. Of 24 MRSA isolates 20 (83.3%) carried MSCRAMMs genes including eno (70.8%), fib (54.1%), cna (25.0%), fnbB (16.6%), ebps 5 (20.8%), and the fnbA, bbp and clfA genes were not detected in any MRSA isolate. MLST analysis revealed 11 sequence types among MRSA isolates as follows: ST239, ST291, ST22, ST861, ST889, ST8, ST59, ST343, ST772, ST6 and ST1465. Also seven MLST-based clonal complexes (CCs) were identified among MRSA strains including: CC8, CC7, CC398, CC59, CC22, CC1 and CC5. CONCLUSIONS: A relatively high diversity was found in MRSA genotypes in Kashan and Isfahan hospitals, and seven clonal complexes were identified. Pandemic MRSA clones including CC8 and CC22 were the most prevalent clones and the novel ST types including ST1465, ST861, ST 889 and ST772 are reported for the first time in Iran in the present study. In addition the high prevalence of MSCRAMMs genes in MRSA isolates demonstrates the high potential of these strains for pathogenicity.

Construction, Cloning, and Expression of CagA Recombinant Protein of Helicobacter pylori.

BACKGROUND: This study aimed to assess construction and expression of CagA recombinant protein of Helicobacter pylori (H. pylori) in Escherichia coli (E. coli) BL21. METHODS: Bioinformatics was used in designing the desired gene by Gene Runner. Next, the construct was subcloned to pET21b vector and this process was confirmed by Polymerase Chain Reaction (PCR), enzyme digestion and sequencing techniques. Then, it was cloned in the Escherichia coli BL21 as an expression host. Expression of protein was verified using sodium dodecyl sulfate- polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting technique. For purification of the protein, the Ni-NTA column was used. Protein concentration was determined by the Bicinchoninic Acid Protein Assay Kit (Parstoos). Finally, Western blotting was performed using CagA antibodies and normal human serum for determining immunogenicity feature with human antiserum. RESULTS: According to the results of the present study, CagA construct was cloned into the pET21b vector and after confirmation and cloning in host expression, recombinant protein with the size of 38 kDa was successfully expressed and purified. The recombinant CagA protein showed immunogenicity characteristics with human antiserum. CONCLUSION: In conclusion, only 5'-end of recombinant protein CagA with high immunogenicity effects was successfully constructed, cloned and expressed. Also, CagA recombinant protein showed good immunogenicity activity with human antiserum.

Environmental soil contamination by Toxocara species eggs in public places of Ilam, Iran.

INTRODUCTION: The purpose of the study is to assess environmental contamination by Toxocara species eggs in public places in the city of Ilam, Ilam Province, southwest Iran. MATERIAL AND METHODS: Between September 2018 and March 2019, 130 soil samples were collected from public places of 5 district municipalities of Ilam, southwest Iran. Soil samples were examined by microscopy following flotation method by sodium nitrate. RESULTS: Soil analysis showed that 5.88% of the soils stored, 52.54% from gardens, 29.42% from rubbish, and 11.72% from green spaces were contaminated with Toxocara spp. eggs. In total, 13.08 % of soil samples (17/130) were positive for Toxocara eggs (P > 0.05). CONCLUSIONS: The findings revealed that care should be taken when using soil from gardens, green spaces and rubbish, and also should be seriously considered because of the potential issues of toxocariasis and also the risk to the public.

Prevalence of Toxocara and Toxascaris infection among human and animals in Iran with meta-analysis approach.

BACKGROUND: Toxocariasis is a worldwide zoonotic parasitic disease caused by species of Toxocara and Toxascaris, common in dogs and cats. Herein, a meta-analysis was contrived to assess the prevalence of Toxocara/Toxascaris in carnivore and human hosts in different regions of Iran from April 1969 to June 2019. METHODS: The available online articles of English (PubMed, Science Direct, Scopus, and Ovid) and Persian (SID, Iran Medex, Magiran, and Iran Doc) databases and also the articles that presented in held parasitology congresses of Iran were involved. RESULTS: The weighted prevalence of Toxocara/Toxascaris in dogs (Canis familiaris) and cats (Felis catus) was 24.2% (95% CI: 18.0-31.0%) and 32.6% (95% CI: 22.6-43.4%), respectively. Also, pooled prevalence in jackal (Canis aureus) and red fox (Vulpes vulpes) was 23.3% (95% CI: 7.7-43.2%) and 69.4% (95% CI: 60.3-77.8%), correspondingly. Weighted mean prevalence of human cases with overall 28 records was 9.3% (95% CI: 6.3-13.1%). The weighted prevalence of Toxocara canis, Toxocara cati, and Toxascaris leonina was represented as 13.8% (95% CI: 9.8-18.3%), 28.5% (95% CI: 20-37.7%) and 14.3% (95% CI: 8.1-22.0%), respectively. CONCLUSION: Our meta-analysis results illustrate a considerable prevalence rate of Toxocara/Toxascaris, particularly in cats and dogs of northern parts of Iran. The presence of suitable animal hosts, optimum climate and close contact of humans and animals would have been the reason for higher seroprevalence rates of human cases in our region. Given the significance clinical outcomes of human Toxocara/Toxascaris, necessary measures should be taken.

Ability of biofilm production and molecular analysis of spa and ica genes among clinical isolates of methicillin-resistant Staphylococcus aureus.

OBJECTIVE: This study aimed to evaluate the phenotypic and genotypic characterization of biofilm formation and spa and ica genes among clinical isolates of methicillin-resistant Staphylococcus aureus. RESULT: This cross-sectional study was performed on 146 Staphylococcus aureus isolates from hospitalized patients in Isfahan Province Hospitals. MRSA isolates were confirmed using disk diffusion test with oxacillin disk and amplification of mecA gene by PCR assays. Ability of biofilm production was evaluated targeting the icaA and icaD genes. Of 146 Staphylococcus aureus isolates, 24 (16.4%) carried mecA genes and identified as MRSA strains. Strong ability of biofilm production was seen among 76.02% (111/146) S. aureus isolates and 87.5% (21/24) MRSA strains, respectively. Also, 75.0% (18/24) MRSA isolates carried icaA and icaD was not detected in these strains. Analysis of spa gene showed 70.83% (17/24) MRSA strains were spa positive. From which 14 and 3 strains identified with one band (150, 270, 300, 360, 400 bp) and two bands (150-300 bp), respectively. According to data obtained, the prevalence of MRSA isolates from Isfahan Province Hospitals is relatively high and a remarkable percentage of them show strong power in biofilm production. Also analysis of spa gene showed a fairly large diversity among MRSA strains.