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Molecular Detection of Common Bacterial Pathogens Causing Meningitis
H Sadighian,MR Pourmand
Iranian Journal of Public Health , 2009,
Abstract: "nBackground: The clinical diagnosis of meningitis is crucial, particularly in children. The early diagnosis and empiric an-tibi-otic treatments have led to a reduction in morbidity and mortality rates. PCR and the enzymatic digestion of 16SrDNA frag-ment which is produced by universal primers led up fast and sensitive determination. The purpose of this study was to investi-gate a rapid method for detection of common bacterial pathogens causing meningitis."nMethods: According to the gene encoding 16SrDNA found in all bacteria, a pair of primers was designed. Then the univer-sal PCR was performed for bacterial agents of meningitis (Streptococcus pneumoniae, Neisseria meningitidis, Haemophilus influ-enzae, etc.) by employing broad- range DNA extraction method. The ob-tained uni-versal PCR products were digested with restriction enzymes (HaeIII, AluI and MnlI) to identify bacterial species. "nResults: By the enzymatic digestion of the universal products of each standard strain of the above bacteria, specific patterns were achieved. These specific patterns may be used for comparison in CSF examination. The analytical sensitivity of the as-say was approximately 1.5′102 CFU/ml of CSF even in samples with high amount of proteins. Conclusion: The universal PCR coupled with enzymatic digestion can be used to detect and identify bacterial pathogens in clini-cal specimens rapidly and accurately. Molecular diagnostic of bacterial meningitis, though expensive and labor-inten-sive, but is valuable and critical in patient management.
Campylobacter Enteritis among Children in Iran
MR Pourmand,MH Shirazi,M Aarabi
Iranian Journal of Public Health , 2005,
Abstract: Campylobacter enteritis is a common form of acute gastroenteritis. Among children, especially in developing countries, Campylobacter infections can cause sever life-threatening diarrheal disease. The incidence of Campylobacter infection among children is age related with a higher incidence among younger children in the developing world whereas in industrialized countries the incidence is highest in older children. In a study of American children, Campylobacter was isolated in 4.8% of diarrheal stools in aged 1-4 years. In 1985 the prevalence of Campylobacter diarrheal was 4.4%, whereas in current report 6% of stool samples from children aged<5 years with diarrhea grew Campylobacter jejuni. There were no significant differences between age groups of patients. All thirteen isolated strains of Campylobacter were resistance to Bactrim, Colistin and Polymyxin B and were sensitive to Neomycin, Erythromycin, Gentamicin and Nalidixic acid. The incidence of human campylobacteriosis is increasing worldwide. Thus, public health awareness about the problem is necessary, with a view towards setting up national surveillance programs.
Characterization of Vancomycin Resistant Enterococcus faecium
M Talebi,SS Eshraghi,MR Pourshafie,MR Pourmand
Iranian Journal of Public Health , 2007,
Abstract: Background: To determine the species distribution, updated drug susceptibility patterns and genes conferring resistance in clini-cal vancomycin resistant enterococcal (VRE) isolates.Methods: Clinical enterococcal isolates collected during 7 months, from September 2005 to April 2006 from hos-pitalized pa-tients and outpatients were studied. Twenty five VRE were isolated from 450 enterococci samples (5.6%). VRE isolates were subjected to antibiotic susceptibility tests. Genotype of these isolates was determined by PCR.Results: All of the isolates were E. faecium and carried the vanA gene. Antibiotic susceptibility tests showed that the iso-lates were resistant to ampicillin 25(100%), ciprofloxacin 25(100%), gentamicin 24(96%), erythromycin 25(100%), tetracy-clin 10(40%) and chloramphenicol 2(8%).Conclusion: VRE strains were resistant to three antibiotics and were susceptible to new antibiotics linezolid and dalfopris-tin-quinupristin. Switching to treatment with these antibiotics would relieve the problem for a short time.
Isolation of Enteroinvasive Escherichia Coli (EIEC) from Diarrhea in Children by Polymerase Chain Reaction (PCR) Technique
A Akbari,MR Pourmand,MK Sharifi Yazdi,M Hosseini
Payavard Salamat , 2010,
Abstract: Background and Aim: Enteroinvasive Escherichia coli (EIEC) strains include a group of diarrheagenic Escherichia coli (DEC) and are known to cause shigellosis-like symptoms in both adults and children. They belong to a limited number of serotypes and their somatic (O) antigens are identical with, or related to, certain Shigella antigens. EIEC strains are confirmed by demonstration of invasiveness by polymerase chain reaction (PCR) for detection of the ipaH (invasive plasmid antigen H) gene that is specific for these strains among DEC.Since in our country,Iran study for detection of these strains. hasnot been carried out therefore the aim of this study was detection of EIEC in diarrheal under 5 year old children in Tehran.Materials and Methods: During the descriptive study,300stool samples were collected from children with diarrhea in Ali Asghar Hospital and children medicinal center of Tehran during 4 months (April-Jul 2008). E.coli species were isolated by standard bacteriological and biochemical tests. Presence of invasive plasmid antigen H (ipaH) gene in confirmed colonies was investigated by PCR technique.Results: Among 300 stool specimens studied using culture method and biochemical tests,39(13%) E.coli species were isolated. Among these 39 strains,7(2.3%) strains containing ipaH gene (EIEC) were detected by PCR technique.Conclusions: Enteroinvasive Escherichia coli (EIEC) in our country, Iran, may be as bacterial pathogen causing childhood diarrhea. Therefore we should apply new techniques for investigation of these strains.
Prevalence of Genes Encoding Bi-Component Leukocidins among Clinical Isolates of Methicillin–Resistant Staphylococcus aureus
SA Havaei,S Ohadian Moghadam,MR Pourmand,J Faghri
Iranian Journal of Public Health , 2010,
Abstract: "nBackground: Staphylococcus aureus has been recognized as a major human pathogen and is the major cause of nosocomial infections. Gamma-toxin, leukocidin and other bi-component toxins are a family of proteins encoded by the hlg and luk-PV, respectively. Panton-Valentine leukocidin (PVL) is an example of these toxins and causes leukocyte destruction and tissue necrosis. The aim of this study was to determine the prevalence of bi-component leukocidin in Methicillin - Resistant Staphylococcus aureus (MRSA) isolates in staphylococcal infections."nMethods: Collectively, 143 isolates of S. aureus were obtained from Tehran University of Medical Sciences hospitals and confirmed with biochemical tests. Then polymerase chain reaction was used to detect luk-PV loci and luk-E/D. Coagulase gene was used as internal control. The antibiotic susceptibility patterns of isolates were determined using disk diffusion method."nResults: Out of 149 S. aureus isolates 24.2% were luk-PV positive and 73.8% were luk-E/D positive."nConclusion: There was PVL-positive MRSA isolates with high prevalence in evaluated hospitals. The diseases from these bacteria are with extensive necrosis, leucopenia and even death. We desire that, prevent from progress and death by diagno-sis and right treatment.
Minimum inhibitory concentration of ciprofloxacin in combination with hexahydroquinoline derivatives against Staphylococcus aureus
M Yousefi,MR Pourmand,AR Shahverdi,M Amini
Tehran University Medical Journal , 2012,
Abstract: Background: Staphylococcus aureus is the most common pathogen responsible for skin and soft tissue infections worldwide. Methicillin-resistant S. aureus is a major cause of both nosocomial and community acquired infections. The emergence of antimicrobial-resistant S. aureus is of global concern. Fluoroquinolone antimicrobials including ciprofloxacin, levofloxacin, and moxifloxacin are used to treat skin and soft tissue infections due to S. aureus. Emergence of ciprofloxacin resistance has increased in community acquired methicillin-resistant S. aureus strains. The aim of this study was to evaluate the minimum inhibitory concentration of ciprofloxacin and hexahydroquino-line derivatives against methicillin- and ciprofloxacin-resistant S. aureus.Methods: Identification of S. aureus was performed by routine microbiological tests in the Department of Pathobiology in Winter 2012. The susceptibility of S. aureus strains to both methicillin and ciprofloxacin was examined by the Kirby-Bauer disk-diffusion method. The minimum inhibitory concentration of ciprofloxacin, hexahydroquinoline derivatives and their combination were separately determined by broth microdilution method against methicillin- and ciprofloxacin-resistant S. aureus.Results: The minimum inhibitory concentration of ciprofloxacin decreased in the presence of hexahydroquinolinein derivatives in comparison with ciprofloxacin alone.Conclusion: This study showed that hexahydroquinoline derivatives enhance the antibacterial effect of ciprofloxacin against methicillin- and ciprofloxacin-resistant S. aureus. Therefore, these derivatives could be used as inhibitors of antibiotic resistance in combination therapies. This enhancement may be related to the inhibitory effect of hexahydroquinoline derivatives on the expression of antibiotic efflux pump in the bacteria. However, the structural features of a fluoroquinolone that determine whether it is affected by efflux transporters are not fully defined.
Prevalence of tst, entC, entA and entA/C genes in staphylococcus aureus strains isolated from different foods
Eshraghi S,Salehipour Z,Pourmand MR,Rahimi Forushani A
Tehran University Medical Journal , 2009,
Abstract: "nBackground: Staphylococcus aureus is a major foodborne pathogen throughout the world. Enterotoxins and toxic shock syndrome toxin-1 are important virulence factors and as pyrogenic toxin superantigens have profound effects on the ir host. Thus circulation of TSST1 producing S.aureus among people and food chain is a worrying issue. The present paper was conducted to study Prevalence of tst, entC, entA and entA/C genes in staphylococcus aureus strains isolated from different foods. "n"nMethods: Over 1040 food samples have been analyzed differentially according to Iran national standard (number= 1194) for S.aureus identification. After DNA extraction, PCR reactions were carried out by reference strain as positive control, adequate primers. "n"nResults: At present study, prevalence of foodstuffs contaminated by S.aureus isolates was about 9.5% (100 strains). Of 25% of isolates producing entC, 28% (seven strains) had tst gene at the same time and of 8% of isolates producing entA, 12.5% (one strain) were positive for tst genes simultaneously. Altogether of 9% isolates producing combination of entC and entA, 44.4% (four strains) were also producer of tst gene. "n"nConclusion: Prevalence of TSST1 producing strains in combination with enterotoxin genes is considerable especially with entC and A plus C. On the other hand, circulation of these isolates in humans, animals, foods and environment has hazardous effect for general public health.
The Identification of Nocardia in BAL Specimens of Bronchoscopic Patients by Using Classical and Molecular Methods
Heidarzadeh S,Pourmand MR,Ghasemi A,Zarrinfar H
Tehran University Medical Journal , 2011,
Abstract: Background: Nocardiosis is a rare and potentially life-threatening infection caused by several species of the Nocardia genus. The objective of this study was to develop and evaluate a rapid and new method to clinically identify relevant Nocardia species. Rapid and accurate diagnosis of Nocardia species is essential for the treatment of severe infections and prevention of cerebral abscess. Methods: One hundred and eighty patients, 103 (57.22%) male and 77 (42.78%) female, with severe symptomatic pulmonary infection were studied in the course of a 12-month period in Dr. Shariati Teaching Hospital affiliated to Tehran University of Medical Sciences in 2010. The specimens were cultured and identified using microbiological and biochemical tests. Polymerase chain reaction (PCR) was used to directly identify the organism in the broncoalveolar lavage samples collected from the patients. NG1 and NG2 primers were used to amplify a Nocardia genus-specific 598-bp fragment of 16S rRNA.Results: Nineteen samples (10.56%) were positive with PCR and 5 samples (2.78%) with conventional methods. All samples with positive cultures were also positive by PCR.Conclusion: The results of this study showed that PCR has a high sensitivity and accuracy for the detection of Nocardia compared with culture and biochemical tests. Considering the rapidity, precision, high sensitivity and specificity of molecular techniques, use of these techniques is suggested in conjunction with conventional methods for the detection of Nocardia phenotypes in clinical laboratories and research centers.
The importance of inducible clindamycin resistance in enterotoxin positive S. aureus isolated from clinical samples
Memariani M,Pourmand MR,Shirazi MH,Soltan Dallal MM
Tehran University Medical Journal , 2009,
Abstract: "nBackground: Clindamycin is a suitable antibiotic for treatment of skin and soft tissue infections. Moreover, it can suppress toxin production in many pathogenic bacteria such as S. aureus. There are two mechanisms of resistance in this antibiotic. Constitutive resistance can be detected by standard disk diffusion method but in the case of inducible resistance, D-test should be carried out. The main aim of this study is to determine prevalence of clindamycin inducible resistance among methicillin resistant and susceptible isolates of S. aureus isolated from different clinical samples. "nMethods: A total of 87 clinical isolates from clinical samples were collected. Methicillin resistance was determined using standard disk diffusion method. Subsequently, D-test was carried out according to CLSI guideline. Presence of the sea gene (enterotoxin A) was detected by PCR using specific primers. "nResults: Out of 87 isolates, 18(20.7%) were clindamycin inducible resistant while constitutive resistance was detected among 21(24.1%) isolates. The 95% Confidence intervals for the proportion of inducible clindamycin resistance among clinical isolates of S. aureus was 12.2% to 29.2%. The inducible phenotype in MRSA isolates was more common than that of MSSA isolates (33.3% vs 5.1%).Significant differences were found between prevalence of inducible clindamycin resistance and type of infection (p=0.045). Importantly, there was a significant correlation between sea gene and the constitutive/inducible resistance (p<0.0001). "nConclusions: Due to the high prevalence of clindamycin inducible resistance among clinical isolates of S. aureus, we recommend D-test to avoid treatment failure.
Evaluating the Efficiency of Lettuce Disinfection According to the Official Protocol in Iran
M Yarahmadi,M Yunesian,MR Pourmand,A Shahsavani
Iranian Journal of Public Health , 2012,
Abstract: Background: The objective of this study was to evaluate the efficacy of Sanitization of Lettuce according to the protocols set forth by Iranian Ministry of Health and Medical Education for reducing populations of total coliform, fecal coliform, and helminth eggs present on lettuce.Methods: In the present study, we determined the load of total coliform, fecal coliform, and parasites of lettuce. The lettuce was sanitized by protocol of Iranian Ministry of Health and Medical Education. The protocol consists of 3 levels to disinfect the fruits and vegetables. The procedure was as follows: first washing stage. The leaves of leafy vegetables washed with tap water, second stage, separation of helminth eggs by 3 to 5 droplets of detergent per liter for 5 min; third stage, disinfection of vegetables by calcium hypochlorite solution (with 200 mg/l free chlorine) for 5 min; and finally the disinfected vegetables were washed with tap water.Results: The average initial levels of total coliform and fecal coliform in the samples were 3.36 log10 cfu/g and 2.31 log10 cfu/g, respectively. Helminth eggs were not detected in any of the samples tested. The efficiency of total coliform and fecal coliform removal were 78.1% (0.75 log10cfu/g) and 79.6% (0.67 log10cfu/g), respectively, after washing. This increased up to 94.8(1.44 log10cfu/g) and 98.5% (1.90 log10cfu/g) after the use of detergent. Chlorine disinfection rose these amounts up to 98.3% (2.18 log10cfu/g) and 100% (2.31 log10cfu/g), respectively.Conclusion: By applying the protocol large parts of microorganisms existing on lettuce have indeed been removed.
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