Virulence genes and antimicrobial resistance pattern in Proteus mirabilis strains isolated from patients attended with urinary infections to Tertiary Hospitals, in Iran.

Background: Proteus mirabilis is a frequent reason for catheter-associated urinary tract infections (UTIs). The aim of this study was to identify virulence genes and antimicrobial resistance patterns in P. mirabilis strains isolated from patients who attended a tertiary hospital in Iran. Methods: In this study, 100 P. mirabilis strains from urine samples were isolated. These isolated strains were identified by biochemical and PCR-based tests, and their antibiotic resistance was profiled through a standard procedure using 14 antibiotics. PCR assays were used to detect virulence-related genes in P. mirabilis strains. The biofilm formation of each P. mirabilis strain was examined. Results: Of the 100 P. mirabilis isolates, 16 (16%) were multidrug-resistant. High resistance was observed against cotrimoxazole (97%), nalidixic acid (93%), cefotaxime (77%), and amoxicillin (62%). Sixty of the 100 isolates showed resistance against extended-spectrum cephalosporins. The prevalence rates of the genes related to the virulence factors in this study were mrpH (100%), ucaA (91%), hpmA (94%), zapA (95%), ptaA (100%), ureG (100%), pmfA (100%), fliC (97%), and mrpA (90%) using PCR method. Strong biofilm formation was observed in 20% (5/25) of the strains isolated from non-catheterized samples and 80% (20/25) of strains isolated from catheterized samples. Conclusions: Resistance to antibiotics and the prevalence of pathogenicity genes are high in Proteus mirabilis strains iolated from UTIs.

Genotypic identification of Pseudomonas aeruginosa strains isolated from patients with urinary tract infection.

BACKGROUND: Numerous nosocomial infections including urinary tract infection (UTI) have been reported to be linked to Pseudomonas aeruginosa (P. aeruginosa). This bacterium is one of the most common pathogen colonized in the urinary tract. The main purpose of this study was to evaluated the presence of antibiotic resistance genes and also the most frequent genotype patterns of P. aeruginosa in the patients with UTI hospitalized in different wards of hospitals. MATERIALS AND METHODS: In this study, 70 strains of P. aeruginosa isolated of urine samples from the patients with UTI were assessed. The isolated strains were genotyped using Multiple-Locus Variable Number Tandem Repeat Analysis (MLVA) method. We have also analyzed the presence of TEM and SHV resistant genes in the isolates. RESULTS: A total of 70 P. aeruginosa strains was isolated from the UTI patients. Based on MLVA method, 61 various genotypes of P. aeruginosa were identified which grouped into two main clusters and 4 sub-clusters. Moreover, approximately 80% and 70% of isolated strains carried the TEM and SHV resistance genes, respectively. CONCLUSION: Our findings showed that the majority of patients hospitalized in different wards of hospitals have experienced the urinary tract infection caused by P. aeruginosa. According to the genotyping results, a high diversity of the P. aeruginosa population was observed in the patients with UTI. Our results can provide a better understanding of the P. aeruginosa genotype distribution and epidemiology of infection, which can be applied as basic data for future antibiotic therapies.

C - reactive protein of cerebrospinal fluid, as a sensitive approach for diagnosis of neonatal meningitis.

BACKGROUND: Meningitis, is a potentially life-threatening condition that can rapidly progress to permanent brain damage, neurologic problems, and even death. Bacteria and viruses cause the great majority of meningitis disease in infants and children. CRP is used mainly as a marker of inflammation. OBJECTIVE: This study was conducted to assess the diagnostic value of CSF-CRP levels for differentiating between septic (bacterial) and aseptic infantile meningitis. METHODS: 49 hospitalized infants aged less than two months with suspected meningitis were enrolled in a cross-sectional analytic study. All of patients underwent lumbar puncture to obtain CSF. smears, cultures, cytological and biochemical analysis and latex agglutination testing were carried out on all CSF samples. Latex agglutination test was carried out on all CSF samples using a commercially available kit. CSF-CRP level of all infants was measured using the immunoturbidometric technique. RESULTS: Of 49 infants in this study, 20 and 29 cases were diagnosed as septic and aseptic meningitis, respectively. The CRP levels were obtained as 0.95±0.68 mg/L in septic and 0.16±0.36 mg/L in aseptic meningitis groups and this difference was statistically significant (p<0.001) between the two groups (0.79±0.32 mg/L). Based on the ROC curve, cut off levels for CRP was obtained 0.17 mg/L. At this level, there was 95% sensitivity and 86% specificity to differentiate septic and aseptic meningitis. CONCLUSION: CSF-CRP has suitable diagnostic value in distinguishing between infantile bacterial from aseptic meningitis especially in cases of negative bacterial culture of the blood and spinal fluid.

Diagnostic Value of CRP, Procalcitonin, and Ferritin Levels in Cerebrospinal Fluid of Children with Meningitis.

BACKGROUND AND OBJECTIVE: Bacterial meningitis is a serious disease with high rate of mortality and morbidity in children. Invasion of pathogens causes brain and meningeal inflammation, which leads to the release of biomarkers into cerebrospinal fluid (CSF). Identification of these biomarkers can help the physicians to differentiate between bacterial and aseptic meningitis. In the current study, some of these biomarkers such as Procalcitonin, C reactive protein (CRP), and Ferritin, were compared in cerebrospinal fluid (CSF) of patients with bacterial and aseptic meningitis. METHODS: In a prospective cross sectional study in a referral children hospital in Tehran during 2011- 2013, the CSF levels of Procalcitonin, Ferritin, and CRP were measured in 57 children with clinically suspected meningitis. The Mann-Whitney u test and the chi-square test were used to compare two groups, children with bacterial and aseptic meningitis. The cut-offs of biomarker levels for differentiation between the 2 groups were constructed by receiver-operating - characteristic curve (ROC). RESULTS: 57 subjects (30 bacterial and 27 aseptic meningitis), were enrolled in this survey. In comparing the two groups, the CSF levels of Ferritin, CRP, and procalcitonin in bacterial meningitis were significantly higher than in aseptic meningitis (P values=˂0.001, 0.001, ˂0.001respectively), with sensitivity/ specificity being 92.9% / 68%, 92.9% / 84%, and 96.4% / 80%, respectively. Positive Predictive Values (PPV) were, correspondingly, 96.4%, 92.8%, and 92.8% for procalcitonin, Ferritin, and CRP. Corresponding Negative Predictive Values (NPV) were, respectively equal to 95.4%, 70%, and 88%. CONCLUSION: CSF of children with bacterial meningitis contains higher levels of inflammatory mediators including Procalcitonin, Ferritin, and CRP, compared to aseptic meningitis The biomarkers provided high sensitivity (especially PCT) and specificity (especially CRP). Using these complementary biomarkers would be useful for early diagnosis of bacterial meningitis and selection of appropriate treatment.

Double-stranded RNA viral infection of Trichomonas vaginalis (TVV1) in Iranian isolates.

The Totiviridae family includes a number of viruses that can infect protozoan parasites such as Leishmania and Giardia and fungi like Saccharomyces cerevisiae. Some isolates of Trichomonas vaginalis are also infected with one or more double-stranded RNA (dsRNA) viruses. In this study, the frequency of Trichomonas vaginalis virus (TVV1) was evaluated in Iranian isolates of T. vaginalis in Tehran, Iran. One thousand five hundred vaginal samples were collected from patients attending obstetrics and gynaecology hospitals associated with Iran University of Medical Sciences in Tehran, Iran from October 2015 to September 2016. Trichomonas vaginalis isolates were cultured in Diamond's modified medium. Nucleic acids were extracted using a DNA/RNA extraction kit and RT-PCR was performed. Among 1500 collected vaginal samples, 8 (0.53%) cases of T. vaginalis infection were found. Half (4/8) of the T. vaginalis positive cases were infected with TVV1. Phylogenetic mapping indicated that the Iranian isolates were most closely related to TVV1-OC5, TVV1-UR1. Iranian isolates of T. vaginalis were infected with TVV1. The frequency of viral infection (TVV1) in T. vaginalis isolates found in this study is higher than previously reported in Iran.

Serum levels of vitamins A and D, and zinc in children with acute diarrhea: A cross-sectional study.

BACKGROUND: Diarrhea is a leading cause of mortality and morbidity during the early life period especially in developing countries. Micronutrients deficiencies have been proposed either as a risk factor or a consequence of diarrhea. Association studies highlight the relation of vitamins and minerals' deficiencies with acute diarrhea. In this regard we aimed to evaluate the status of vitamins A and D, and zinc serum levels in children with acute diarrhea. METHODS: In this cross sectional study performed in a referral teaching hospital, we measured and compared baseline vitamin A, 25-hydroxy vitamin D (25(OH)D), and zinc serum levels in 25 children admitted with acute diarrhea and 25 other children who were admitted for undergoing elective surgeries. RESULTS: 25-(OH)D levels were significantly lower in the diarrhea group (p=0.03). We were unable to demonstrate a significant difference in the levels of vitamin A and zinc between the case and control groups (p= 0.14 and p=0.07, respectively). CONCLUSION: We observed lower serum 25(OH)D levels in children with acute diarrhea. Whether this finding indicates a premorbid risk factor or simply a consequence of diarrhea needs further studies. Regardless of the cause and effect relationship, supplementation with vitamin D in acute diarrhea remains as a plausible consideration.

Searching the Staphylococcal Superantigens: Enterotoxins A, B, C, and TSST1 in Synovial Fluid of Cases With Negative Culture Inflammatory Arthritis.

BACKGROUND: Accurate and rapid diagnosis of bacterial arthritis is not always possible in unvaccinated (Streptococcus pneumoniae and Haemophilus influenzae type B) children in Iran. OBJECTIVES: Searching the staphylococcal superantigen (entrotoxin A, B, C and TSST1) in synovial fluid of cases with inflammatory arthritis. PATIENTS AND METHODS: This cross sectional study was implemented in the pediatric and orthopedic wards, Rasoul Akram Hospital, Tehran, Iran (2008-2010) upon synovial fluid (SF) aspirated from 66 children (five months to 16 years; mean age 11 ± 3.8 years) with monoarthritis. Staphylococcal supperantigens (enterotoxins A, B, C, TSST1) were assessed by Enzyme-linked immunosorbent assay (ELISA) in synovial fluid of cases with inflammatory arthitis. Staphylococcal superantigens compared between cases with positive and negative Staphylococcus aureus culture (P < 0.05 was significant). RESULTS: S. aureus was the most common cause of septic arthritis. Positive S. aureus culture in SF was reported in 10.6% (7/66) of the cases. Enterotoxin A was the least common type of superantigens found even in SF negative culture; 47% of the cases had one or more staphylococcal superantigens. Enterotoxin A was the least common type in SF; there was poor agreement between positive culture for S. aureus and presence of enterotoxins B, C, and TSST1 in SF, and intermediate agreement (KAPPA Index = 0.67) for enterotoxin A. CONCLUSIONS: A possible role (%47) for staphylococcal toxins was defined even in SF negative cultures obtained from monoarthritis cases. Failure in isolation of organisms might be due to natural un-growth of microorganism in synovial fluid, and previous antibiotic usage or low technical methods. It could not be determined from the data obtained in the current investigation whether or not staphylococcal toxins (superantigens) play a pathogenic role without direct invasion of the organism. It is recommend to use complementary methods for searching the S. aureus superantigens in future studies.

The rise of staphylococcal super antigens in psoriatic patients: a case-control study.

BACKGROUND: Staphylococcus aureus, the major virulence factor of hospital and community acquired infections, secretes numerous exotoxins (super antigens), which may affect immunological and inflammatory status in psoriatic skin lesion. OBJECTIVES: This study is designed to compare the S. aureus super antigens level in sera of psoriatic patients with normal cases (nevus). PATIENTS AND METHODS: A case control study was performed in dermatology ward of Rasoul Hospital in Tehran, IR Iran (2008 - 2010). Staphylococcal super antigens (Entrotoxin A, B, D and TSST1) were measured in serum of 41 psoriatic patients and 28 normal persons (Nevus) by ELISA. Chi square values (CI 95%, P value < 0.05) were calculated for all categorical variables. RESULTS: In this study 63.4% (26) of cases were male, 36.6% (15) were female. Age ranged from 4 months to 64 years old, with a mean age of 33.7 ± 15.4 years. Type of skin disease in cases: 20% (8) were inflicted by the Gutate form of the disease; 59% (23) with chronic plaque psoriasis (CPP), 7.7% (3) with erythrodermic and 12.8% (5) had other types of the disease (plaque, pustular, inverse). TSST (toxic shock syndrome toxin) was detected in 47% (20/41) of cases and in 6% (1/28) of the controls with a significant difference. (P value = 0.000) Entrotoxins (A, B, D) were detected in the sera of 48.8% (21/41) of cases; and only 6 %( 1/21) of controls, showed significant differences (P value = 0.000) positive TSST was more common in spring, and correlates with CPP type of psoriasis, but not related to patient`s gender and age. CONCLUSIONS: In this study, S. aureus were 25 times more in psoriatic patients. Super antigens should be first detected in the serum samples; if negative, the skin lesions should be examined by PCR especially in chronic types of disease. Adding the antibiotics against S. aureus to other conventional treatments might be helpful. It has a more important and significant role in children with acuteinfection.

Assessments of blood lead levels in children with febrile convulsion.

BACKGROUND: Lead elements have an adverse effect on human health. The most important complications of lead poisoning are disorders of nervous system particularly seizure .This study aimed to evaluate the blood lead levels and its association with convulsion in a group of hospitalized febrile children. METHODS: In this analytic cross-sectional study, 60 hospitalized febrile children with 1- 60 month old participated in the study via non-probability convenience sampling method. All of the information included sex, age, weight, blood lead levels and history of convulsion gathered in the questionnaire. Finally all of data were statistically analyzed. RESULTS: 66.7% of samples were male and 33.3% were female. The mean age was 32.57±38.27 months and the mean weight was 13.04±9.61kg. The Mean and Standard deviation of Blood lead level was 4.83±3.50µg/dL. 10% of samples had lead levels greater than 10µg/dL. 53.3% of patients have convulsion and other don't have it. Blood lead levels was 4.91±3.65µg/dL in children with convulsion and 4.73± 3.38µg/dL in children without it; the difference was not significant (p= 0.8). CONCLUSION: Overall, no significant association was found between blood lead levels and convulsion.

Searching the H. pylori; serology & PCR in children with adenoid hypertrophy and rhino sinusitis: a cross sectional study, Tehran, Iran.

OBJECTIVE: Searching the H. pylori -DNAs in adenoid and IgA, IgG antibodies in serum of children. METHODS: A cross- sectional study had done upon 53 children (Mean age 8 ±1.9 years) with adenoid surgery in ENT and Pediatric Department of Rasul Akram Hospital (2008-2010). Of 53 cases with adenoid surgery, 40 cases had rhino sinusitis (in sinus CT scan). The resected adenoid tissues (n=40) centrifuged and homogenized and its DNAs extracted and searched for H. pylori- DNAs by qualitative PCR. Serum H. pylori antibodies (IgG &IgA ELISA) calculated quantitatively. Chi square values (p < 0.05) calculated for all categorical variables. The agreement between serologic test and PCR was assessed by the calculation of kappa statistic. RESULTS: Positive PCR for H. pylori detected in 15%of cases; regardless of sex and age of cases. Positive IgA: 17.5% (7/40) and IgG: 20% (8/40) respectively. All cases with positive H. pylori- IgG were female; p=0.003 and meaningful differences in mean age of cases observed for positive IgA (p=0.001) and IgG (p=0.01). Poor agreement observed between positive PCR and serum IgG&IgA (Fisher's Exact test=0.3; 0.5). CONCLUSION: Positive PCR in adenoid tissue (15%) was very close to positive serum IgA (15%) but without any agreement for each case. The H. pylori infection may have a relative role at least in 15% of children with adenoid surgery. Chronic sinusitis and ear infection might be added to infected adenoid tissue as a reservoir for bacteria. The search by specific culture may elucidate better the role of H. pylori infections in both gastric and adenoid tissues. The decision for use of antibiotics to eradicate the H. pylori infection in recurrent or chronic adenotonsillar infections (with rhinosinusitis) before adenoid surgery needs Randomized Control Trial (RCT) studies. Drug of choice for eradication of H. pylori dependent to antibiotic sensitivity test in each country.

Identification of bacterial antigens and super antigens in synovial fluid of patients with arthritis: a cross sectional study.

BACKGROUND: An accurate and prompt diagnosis of bacterial arthritis is essential for earlier treatment and a good outcome. Superantigens produced by Staph. Aureus are among the most lethal toxins. The paper objective was Identification of common bacterial antigens and S.aureus superantigens in synovial fluid (SF) of children with negative culture and direct smear for other bacteria except for S.aureus. METHODS: In this cross-sectional study a total of 62 patients with a mean age of 11 ± 3.8 years (range: 5 months-16 years) with acute arthritis in pediatric and orthopedic wards of Rasoul Hospital (2008-2010) were studied. Three common bacterial antigens (e.g. S.pneumonia, H.influenza, N. meningitis) using LPA (latex particle antigen) and Staphylococcal superantigens (TSST1; Enterotoxin A; B; C) using ELISA method (ABcam; USA) were identified in 60 adequate SF samples with negative culture and negative direct smears) for other bacteria except for S.aureus. Staphylococcal superantigens were compared with S.aureus infection (positive culture or direct smear). RESULTS: Positive bacterial antigens (LPA test) were found in 4 cases including two S. Pneumonia, one N.meningitis, and one H.influenza. S.aureus was diagnosed in 7 cases including 4 positive cultures and 3 positive smears. Staphylococcal superantigens (toxins) were found in 73% of SF samples. Some cases had 2 or 3 types of toxins. S.aureus toxins were reported in 47% of culture negative SF samples. Positive TSST1, Enterotoxin B, Enterotoxin A, and Enterotoxin C were found in 47% (n= 28), 18% (n= 10), 39% (n= 22), and 39% (n = 21) of cases respectively. The most common type of superantigens was TSST1; and Enterotoxin A was the less common type. Except for Enterotoxin A, no relation between positive S.aureus culture and positive tests for superantigens in SF was found. CONCLUSION: S.aureus has a prominent role in septic arthritis. S.aureus toxins might have a prominent role in arthritis with negative SF culture. Rapid identification of bacterial antigens (LPA) or S.aureus superantigens (toxins) are valuable for diagnosis in cases with negative cultures. We recommend usage of complementary methods (e.g. antigen detection tests) in children. Those tests are cheaper and easier in comparison with PCR as a complex and time-taking method. Identification of S.aureus superantigens in SF of all cases with negative culture, or treatment with antagonist drugs needs further clinical trial studies.

Searching Mycoplasma pneumonia by serology & PCR in children with adenoid hypertrophy and rhinosinusitis: A case control study, Tehran, Iran.

BACKGROUND AND OBJECTIVES: Chronic infection in childhood is a leading cause of adeno-tonsillectomy. The aim of this study was to determine the role of M. pneumoniae in children with rhino sinusitis and adenoid hypertrophy. METHOD AND MATERIAL: This case - control study was carried out in the pediatric and ENT wards of Hazrat Rasul Hospital, Tehran, Iran (2007-2009). In this trial, we investigated 40 cases with adenoid surgery and 32 controls.We looked for M. pneumoniae -DNA (PCR) in adenoid tissues resected from cases and 31 nasopharyngeal swabs in controls and IgM & IgG antibodies (ELISA) were compared between the 2 groups, P < 0.05 was considered to show a significant value. RESULTS: Positive PCR results were observed in 35% of cases and none of controls, positive-IgG was seen in 20% of cases and 6.4% of controls (P = 0.71) which was higher in older cases (6 vs. 4 years, p < 0.05). Positive -IgM was seen in 10% of cases vs. 9.7% of controls, (P= 0.74); without any difference for age (6.2/ 5.3 years, p = 0.1). A positive PCR result was not related to positive IgG (p = 0.014), but to a positive IgM (p= 0.1). CONCLUSION: M. pneumoniae infection was found serologically (IgM & IgG) in10% and 20% of cases, respectively. These numbers along with positive PCR in adenoid tissue of cases (30%) indicates the prominent role for M. pneumoniae in adenoid hypertrophy. We concluded that children in Iran will have been infected with M. pneumoniae and would have obtained immunity between the ages of 6 and 8. Adenoid tissue might act as a reservoir for M. pneumoniae and cause rhino sinusitis concomitant with adenoid hypertrophy in infected children. Theoretically, suitable M. pneumoniae eradicating antibiotics before adenoid surgery (with rhino sinusitis or chronic ear infection) might be helpful treatment, but it needs future RCT studies to be proven.

Evaluation of the endothelial cell antibodies in serum and perilymphatic fluid of cochlear implanted children with sensorineural hearing loss.

INTRODUCTION: Serum Anti endothelial Cell Antibodies (AECAs) play a prominent role in idiopathic Sensorineural Hearing Loss (SNHL) in that they induce vascular damage (immune mediated). The of the current study is To compare AECAs in serum and perilymphatic fluid of idiopathic SNHL children (<15y) undergoing cochlear implant surgery. METHODS: This was a cross sectional study performed in the cochlear implant ward in Rasoul Akram hospital, Tehran, Iran (2008 -2010) on 99 SNHL children undergoing cochlear implant surgery. The data collected from47 idiopathic and 52 non-idiopathic SNHL cases. AECAs were measured by indirect immuno fluorescence assay and compared in sera and perilymphatic fluids between the two groups. P-value < 0.05 was considered significant. RESULTS: Idiopathic SNHL was diagnosed in 47.5% of cases. Positive AECA results in serum and perilymphatic fluid were 10% and 12%, respectively. Although AECA results in perilymphatic fluids were different between idiopathic and non-Idiopathic SNHL patients (PV < 0.05), AECAs in serum showed no significant difference between the two (PV = 0.1). No significant difference was detected between the mean age of idiopathic and non-idiopathic SNHL patients with positive AECAs in serum and perilymphatic fluids (PV = 0.2; PV = 0.2). DISCUSSION: Idiopathic SNHL was diagnosed in 47.5% of studied cases. Idiopathic SNHL has a poor out come in children. In cases with idiopathic SNHL, finding AECAs in perilymphatic fluids are more valuable than in the serum. We suggest that serum and perilymphatic fluids testing for AECAs would be helpful in management of idiopathic SNHL cases. Specific immunosuppressive treatments for selected cases suffering from Idiopathic SNHL (only in those older than 5) might be successful in disease management. However, this theory should first be validated by randomized clinical trials.

Infection in childhood sensory hearing loss.

OBJECTIVE: To evaluate the possible role of infectious agents on the occurrence of idiopathic sensorineural hearing loss (SNHL) in children. METHODS: This case control study was carried out at Rasul Hospital in Tehran, Iran from 2002-2003. We compared specific serum antibodies (IgG, IgM) measured by enzyme linked immunosorbent assay in 95 sensory hearing loss cases and 63 controls. RESULTS: Acute infections (IgM) detected in cases included: cytomegalovirus (CMV) 34.6%, toxoplasma 11.5%, mumps 8.7%, rubella 5.3%, and herpes simplex 5.3%. Previous infections (IgG) detected in cases included CMV 72%, herpes 6.6%, toxoplasma 26%, mumps 23.3%, and rubella 17.2%. Acute CMV and toxoplasma infections were more frequent in cases. Previous CMV, toxoplasma, rubella, and herpes infections were higher in controls. There was no significant difference for acute mumps, rubella, and herpes infections between cases and controls. CONCLUSION: These data are compatible with infectious agents having a significant role in the studied idiopathic SNHL cases, but association does not prove causation. We recommend specific drugs for confirmed active infections (CMV, toxoplasma, herpes) in idiopathic SNHL infants diagnosed before their first birthday. Mumps and rubella induced SNHL are preventable with routine vaccination.