Drug resistance patterns of bacteria isolated from patients with nosocomial pneumonia at Tehran hospitals during 2009-2011

INTRODUCTION: Nosocomial pneumonia remains an important cause of mortality and morbidity worldwide. Surveillance programs play an important role in the identification of common etiologic agents and local patterns of antimicrobial resistance. METHODOLOGY: In this study we determined the frequency and antimicrobial susceptibility of pathogens isolated from patients with nosocomial pneumonia during 2009 to 2011. RESULTS: A total of 642 bacteria were isolated from 516 suspected samples. Acinetobacter baumannii (21.1%, n = 136), was the commonest isolated pathogen followed by Pseudomonas aeruginosa (17.4%, n = 112), Staphylococcus aureus (15.8%, n = 102) and enterococci (8.4% n = 54). The most effective therapeutic agents against A. baumannii were polymyxin B (95.5% susceptible), ceftriaxone/tazobactam (72% susceptible) and levofloxacin (52.9% susceptible). Polymixin B (89.2% susceptible), ceftriaxone/tazobactam (89.2% susceptible) and piperacillin-tazobactam (80.3% susceptible) were found to be the most active agents against P. aeruginosa. Extended-spectrum beta-lactamases were detected among isolates of K. pneumoniae (45.4%) and E. coli (20.3%). Overall, the prevalence of methicillin-resistant S. aureus and vancomycin resistant enterococci were 80.4% and 40.7% respectively. Linezolid was found to be the most active antibiotic against these pathogens. The etiology of 50% of the nosocomial infection cases was polymicrobial. CONCLUSIONS: The combination of ceftriaxone/tazobactam seems to be beneficial agent against multidrug-resistant Gram-negative bacilli isolated form respiratory tract infections. The results of our study can be used for guiding appropriate empiric therapy in this geographic region

Development of a modified DNA extraction method for pulsed-field gel electrophoresis analysis of Staphylococcus aureus and enterococci without using lysostaphin

A modified pulsed-field gel electrophoresis (PFGE) protocol was developed and applied to clinical isolates of Staphylococcus aureus and enterococci to reduce the cost of using lysostaphin. This protocol reduces the expenses of PFGE typing of S. aureus and enterococci as it removes the use of lysostaphin during the spheroplast formation from these bacteria

Detection and quantification of Streptococcus pneumoniae from Iranian patients with pneumonia and individual carriers by real time PCR

The aim of this study was to develop a real time polymerase chain reaction (PCR) for quantitative detection of Streptococcus pneumoniae from clinical respiratory specimens. Initially, 184 respiratory specimens from patients with community acquired pneumonia (CAP) (n = 129) and 55 cases with hospital associated pneumonia (HAP) were bacteriologically investigated. To check the colonization status among the healthy individuals, 32 preschool and 31 adults were screened in parallel. All specimens were cultured on selective culture media to isolate S. pneumoniae, Legionella spp. and Mycoplasma spp. A 166 bp fragment corresponding to cbp A gene of S. pneumoniae was amplified from clinical specimens using Taqman probe real time PCR. Culture showed 14, but real time PCR showed 15 specimens as being positive for S. pneumoniae. The specificity and sensitivity of real time PCR was 99.14% and 100 respectively. Co-infections of S. pneumoniae with Legionella pneumophila, Chlamydophila pneumoniae, Mycoplasma pneumoniae and Staphylococcus aureus were observed in 5 cases (35.72%). S. pneumoniae was counted <103 cfu/ml from the co-infected cases. Using real time PCR, a cutoff of 103cfu/ml is introduced to differentiate colonization from infection in respiratory tract. This is the first report on the prevalence CAP with S. pneumoniae in Iran (12.40%).Key words: Streptococcus pneumoniae, community acquired pneumonia (CAP), real time polymerase chain reaction (PCR), choline binding protein A (cbp A)

Rapid, cost-effective, sensitive and quantitative detection of Acinetobacter baumannii from pneumonia patients

BACKGROUND AND OBJECTIVES: Pneumonia with Acinetobacter baumannii has a major therapeutic problem in health care settings. Decision to initiate correct antibiotic therapy requires rapid identification and quantification of organism. The aim of this study was to develop a rapid and sensitive method for direct detection of A. baumannii from respiratory specimens. MATERIALS AND METHODS: A Taqman real time PCR based on the sequence of bla(oxa-51) was designed and used for direct detection of A. baumannii from 361 respiratory specimens of patients with pneumonia. All specimens were checked by conventional bacteriology in parallel. RESULTS: The new real time PCR could detect less than 200 cfu per ml of bacteria in specimens. There was agreement between the results of real time PCR and culture (Kappa value 1.0, p value<0.001). The sensitivity, specificity and predictive values of real time PCR were 100%. The prevalence of A. baumannii in pneumonia patients was 10.53 % (n=38). Poly-microbial infections were detected in 65.71% of specimens. CONCLUSION: Acinetobacter baumannii is the third causative agent in nosocomial pneumonia after Pseudomonas aeroginosa (16%) and Staphylococcus aureus (13%) at Tehran hospitals. We recommend that 104 CFU be the threshold for definition of infection with A. baumannii using real time PCR

Single tube real time PCR for detection of Streptococcus pneumoniae, Mycoplasma pneumoniae, Chlamydophila pneumoniae and Legionella pneumophila from clinical samples of CAP

We designed a multiplex real time PCR for rapid, sensitive and specific detection of Streptococcus pneumoniae, Legionella pneumophila, Chlamydophila pneumoniae and Mycoplasma pneumoniae. The study cases consisted of 129 patients with community acquired pneumonia (CAP). Bacteriological techniques were implemented for detection of the cultivable organisms. DNA were extracted from sputa, throat swabs, bronchoalveolar lavages and tracheal aspirates and used as templates in real time PCR. The primers and probes were designed for cbpA (S. pneumoniae), p1adhesin (M. pneumoniae), mip (L. pneumophila) and ompA (C. pneumoniae). After optimization of real time PCR for every organism, the experiments were continued in multiplex in a single tube. Of 129 CAP specimens, the positive cultures included 14 (10.85) for S. pneumoniae, 9 (6.98) for L. pneumophila and 3 (2.33) for M. pneumoniae. Four specimens (3.10) were positive for C. pneumoniae by real time PCR. The sensitivity of our real time PCR was 100 for all selected bacteria. The specificity of the test was 98.26, 98.34, 100 and 100 for S. pneumoniae, L. pneumophila, M. pneumoniae and C. pneumoniae, respectively. This is the first report on the use of multiplex real time PCR for detection of CAP patients in the Middle East. The method covers more than 90 of the bacterial pathogens causing CAP. © 2012 Akadémiai Kiadó, Budapest

Community-acquired pneumonia related to intracellular pathogens

Community-acquired pneumonia (CAP) is associated with high rates of morbidity and mortality worldwide; the annual incidence of CAP among adults in Europe has ranged from 1.5 to 1.7 per 1000 population. Intracellular bacteria are common causes of CAP. However, there is considerable variation in the reported incidence between countries and change over time. The intracellular pathogens that are well established as causes of pneumonia are Legionella pneumophila, Mycoplasma pneumoniae, Chlamydophila pneumoniae, Chlamydophila psittaci, and Coxiella burnetii. Since it is known that antibiotic treatment for severe CAP is empiric and includes coverage of typical and atypical pathogens, microbiological diagnosis bears an important relationship to prognosis of pneumonia. Factors such as adequacy of initial antibiotic or early de-escalation of therapy are important variables associated with outcomes, especially in severe cases. Intracellular pathogens sometimes appear to cause more severe disease with respiratory failure and multisystem dysfunction associated with fatal outcomes. The clinical relevance of intracellular pathogens in severe CAP has not been specifically investigated. We review the prevalence, general characteristics, and outcomes of severe CAP cases caused by intracellular pathogens

Serological Prevalence of Toxoplasmosis in Meat Producing Animals

Background: Toxoplasmosis is a zoonotic disease, caused by cosmopolitan coccidian Parasite, Toxoplasma gondii. One of the most important sources of human infection is feeding from raw or uncooked meat of infected animals. In this study the prevalence of toxoplasmic infection in three important meat producing animals in Iran was studied. Methods: Using indirect immunoflourcent antibody assay (IFA), 483 serum samples of goats, sheep and cattle from industrial slaughterhouse of Kermanshah, western Iran were tested for total antibodies against T. gondii. Results: Antibodies to T. gondii were found in 23.7% of goats, 22.5% of sheep and 4.8 % of cattle at titer of ≥1:20. The highest titers observed in goats, sheep and cattle were 1: 2560, 1: 1280 and 1: 640, respectively. Conclusion: It is suggested that feeding of raw or undercooked meat of goats and sheep is important in transmition of toxoplasmic infection to human in Kermanshah district

Detection and quantification of Streptococcus pneumoniae from Iranian patients with pneumonia and individual carriers by real time PCR

The aim of this study was to develop a real time polymerase chain reaction (PCR) for quantitative detection of Streptococcus pneumoniae from clinical respiratory specimens. Initially, 184 respiratory specimens from patients with community acquired pneumonia (CAP) (n = 129) and 55 cases with hospital associated pneumonia (HAP) were bacteriologically investigated. To check the colonization status among the healthy individuals, 32 preschool and 31 adults were screened in parallel. All specimens were cultured on selective culture media to isolate S. pneumoniae, Legionella spp. and Mycoplasma spp. A 166 bp fragment corresponding to cbp A gene of S. pneumoniae was amplified from clinical specimens using Taqman probe real time PCR. Culture showed 14, but real time PCR showed 15 specimens as being positive for S. pneumoniae. The specificity and sensitivity of real time PCR was 99.14 and 100 respectively. Co-infections of S. pneumoniae with Legionella pneumophila, Chlamydophila pneumoniae, Mycoplasma pneumoniae and Staphylococcus aureus were observed in 5 cases (35.72). S. pneumoniae was counted &lt;10 3 cfu/ml from the co-infected cases. Using real time PCR, a cutoff of 10 3cfu/ml is introduced to differentiate colonization from infection in respiratory tract. This is the first report on the prevalence CAP with S. pneumoniae in Iran (12.40). © 2011 Academic Journals

Hospital acquired pneumonia: Comparison of culture and real-time PCR assays for detection of Legionella pneumophila from respiratory specimens at Tehran Hospitals

Legionella pneumophila is an important etiological agent in both hospital and community acquired pneumonia. The sensitivity of culture for isolation of L. pneumophila from clinical specimens is low and time consuming. Similar problem also exists when the method of direct immunofluorescence is used. To detect this organism quantitatively from respiratory specimens, a Taq Man based real-time PCR targeting the mip sequence was developed. Both real-time PCR and culture methods were applied on 262 respiratory specimens from 262 ICU patients with pneumonia admitted to 5 different hospitals in Tehran. The results of real-time PCR were compared with those obtained by culture. Real-time PCR and culture found 12 and 4 specimens, respectively, as positive for L. pneumophila. Its technical specificity (100%) was checked against a panel of microorganisms consisting of both Gram-positive and Gram-negative bacteria. Our real-time PCR assay showed high sensitivity (100%) and specificity (96.9%) and could detect 200 organisms per ml from respiratory specimens. Using real-time PCR as a screening method, the frequency of nosocomial pneumonia with L. pneumophila at Tehran hospitals was estimated as 4.58%