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Comparison of Real-time PCR method and blood culture in diagnosis of septicemia
Ali Gholami , Mohammad Reza Arabestani ,
Volume 73, Issue 11 (2-2016)
Abstract

Background: Bloodstream infections (BSI) have a high incidence and high mortality in the worldwide. The mortality rate is variable between 20-70%. Therefore, early and timely detection of BSI agent in clinical laboratories is necessary. The aim of this study was to determine an efficient diagnostic tool to septicemia in accompany of blood culture method by Real-time PCR (using panbacterial 23S rRNA gene).

Methods: This cross-sectional study was conducted in two analytical and clinical stages in Hamadan University of Medical Sciences, Iran, from October 2014 to June 2015. In analytical stage, sensitivity (by serial dilution from 104 to 1 CFU/ml) and specificity of the primer were evaluated with the Staphylococcus aureus (as Gram positive indicator bacteria) and Escherichia coli (as Gram-negative indicator bacteria), human genome (from Hella cell culture), Candida albicans yeast and Aspergillus fumigatus fungus. In clinical stage, 121 blood samples were collected from patients suspected to sepsis in intensive care unit (ICU) from Hamadan University Hospitals. Finally, the results of Real-time PCR and blood culture methods were compared.

Results: The Real-time PCR showed a sensitivity ranging from 2 to 10 target copies per reaction to the whole blood for Escherichia coli and Staphylococcus aureus respectively. The specificity of this method was evaluated and no false positive amplification was identified. 57.85% (70 cases) of the samples were positive by Real-time PCR and 13.22% (16 cases) of the samples were positive by blood culture. However, none of the cases that were positive by blood culture were negative in Real-time PCR. As well as, 44.62% (54 cases) of cases were positive by Real-time PCR but blood culture showed no bacteria in the samples, and 42.15% (51 cases) were negative by both methods. Correlation or agreement of Kappa was 0.20, that indicating poor agreement between the two methods.

Conclusion: Real-time PCR is more sensitive than blood culture and also, because of high sensitivity of this primer by Real-time PCR, we can use it for screening blood samples from suspected patients of sepsis.


Frequency and antibiotic resistance patterns of isolated bacteria from positive blood culture of hospitalized patients
Azadeh Vahedi , Akram Baghani , Zohre Baseri , Mohammad Reza Pourmand ,
Volume 75, Issue 12 (3-2018)
Abstract
Background: Bloodstream infections are the most important causes of morbidity and mortality in hospitalized patients. Blood culture plays an important role in identifying most of bacterial agents of bloodstream infections. Knowledge about bacterial agents of bloodstream infections and also antibiotic resistance of these bacteria are important. Antibiotic resistance among bacterial agents of bloodstream infection including Acinetobacter, Klebisella, Pseudomonas, Escherichia coli, Enterobacter, Enterococcus, Staphylococcus aureus and Staphylococcus coagulase negative (CoNS) is one of the major challenges faced by physicians in treating. Therefore, this study was aimed to determine the frequency and antibiotic resistant patterns of bacterial isolates from hospitalized patient's blood cultured samples in the hospital, Tehran, Iran.
Methods: This research is a descriptive and retrospective study based on recorded data in Shariati hospital laboratory and under the supervision of Tehran University of Medical Sciences. The bacterial isolates were collected from positive blood cultures from October 2013 to March 2014. The frequency of bacterial isolates were determined by phenotypic and biochemical tests. The antibiotic resistance patterns of isolated bacteria were found by disk diffusion agar method. The diameters of inhibition zone were recorded and interpreted according to Clinical and Laboratory Standards Institute (CLSI) 2013.
Results: The frequency of bacterial isolates was determined among 595 positive blood cultures as followed: 41% Pseudomonas, 20% Staphylococcus epidermidis, 10% Escherichia coli, 6% Acinetobacter lwoffii, 6% Staphylococcus aureus, 5% Stenotrophomonas, 3% Acinetobacter baumannii. The antibiogram test showed that 96.2% of Acinetobacter lwoffii, 92.8% of Acinetobacter baumannii, 66% of Pseudomonas aeruginosa, 85.7% of Staphylococcus epidermidis, 65% of Staphylococcus aureus, 75% of Klebsiella, 73.7% of Escherichia coli, and 50% of Stenotrophomonas were resistant to imipenem, piperacillin, piperacillin, erythromycin, erythromycin, ciprofloxacin, trimethoprim-sulfamethoxazole, and ceftazidime respectively.
Conclusion: The most prevalent bacterial isolate among the blood cultures of patients was Pseudomonas. The patients more than 50 years were more susceptible to blood stream infections. The most bacteria were isolated from the internal medicine department of hospital. The antibiotic resistance was also increasing especially in Acinetobacter, Staphylococcus coagulase negative, Escherichia coil and Klebsiella

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