Vancomycin utilization evaluation at hematology-oncology ward of a teaching hospital in Iran

The aim of the present study was to evaluate the pattern of vancomycin administration in the hematology-oncology ward of Nemazee Hospital, Shiraz, Iran. Study criteria were developed to assess the several parameters involved in vancomycin therapy. These parameters include the appropriateness of drug usage, dosage, duration of therapy, monitoring for toxicity and serum concentration monitoring. The serum concentration was measured by an automated Fluorescence Polarization Immunoassay. Clinical and preclinical parameters such as Glomerular Filtration Rate [GFR], microbial culture, antibacterial sensitivity, WBC count and fever were collected and recorded for analysis. Sixty patients were enrolled in the study, consisting of 45 males and 15 females. The age range was 15 to 68 years. In this study, 68.63% of the vancomycin used for the patients with febrile neutropenia was compatible with the Infectious Disease Society of America [IDSA] guideline. The initial dosage of vancomycin in 68.63%, rate of infusion in 100%, and dilution of vancomycin in100%, were appropriate. Inappropriate use was more evident in the continuation of vancomycin in 50% of the patients. No appropriate dosage adjustment was done for 50% of the patients with increased serum creatinine. Based on the results, the indication of vancomycin in febrile neutropenia was satisfactory. However, there were some required factors such as continuation of vancomycin, adjustment of dosage or interval, microbial culture, antibiotic sensitivity test before the first dose administration, measurement of serum concentration and monitoring which had to be revised in order to achieve an effective treatment

Antifungal susceptibility of the Aspergillus species by Etest and CLSI reference methods

Because resistance to antifungal drugs is seen in patients, susceptibility testing of these drugs aids in choosing the appropriate drug and respective epidemiology. This study has investigated and compared susceptibility patterns of the Aspergillus species sisolated from patients by the Clinical and Laboratory Standards Institute [CLSI] reference broth microdilution [MD] assay and Etest method. The minimum inhibitory concentrations [MICs] of various antifungal agents [amphotericin B, ketoconazole, itraconazole, and voriconazole] for 108 Aspergillus species isolated from patients were determined by CLSI M38-A broth MD and Etest. The isolates were obtained from clinical samples that included tissues, sputum, bronchoalveolar lavage, abdominal tap, and cerebrospinal fluid. As revealed by the MD method, 63.9% of the isolates were sensitive to amphotericin B and 36.1% were resistant. Etest revealed that 61.1% were sensitive to amphotericin B and 38.9% were resistant. As for ketoconazole, 108 isolates [100%] were shown to be sensitive through the MD method; while the Etest revealedan 88.9% sensitivity and 11.1% were resistant. All species were susceptible to voriconazole, according to both methods. The measure of agreement [Kappa Index] for these three drugs was satisfactory [>/= 0.6]. According to the MD method, 69.4% of the species were susceptible to itraconazole, whereas 30.6% were not. For this drug, the Etest showed 86.1% susceptible and 13.9% resistant. Voriconazole was the most effective agent against isolates. Using RPMI agar, we found the Etest to be helpful, readily available, and easy to use for determining invitro susceptibilities of Aspergillus species to voriconazole, amphotericin B, ketoconazole, and itraconazole in the region of this study

Microbial susceptibility, virulence factors, and plasmid profiles of uropathogenic Escherichia coli strains isolated from children in Jahrom, Iran

Background: Urinary tract infections [UTIs], including cystitis and pyelonephritis, are the most common infectious diseases in childhood. Escherichia coli [E. coli] accounts for as much as 90% of the community-acquired and 50% of nosocomial UTIs. Therefore, identification of E. coli strains is important for both clinical and epidemiological implications. Understanding antibiotic resistance patterns and molecular characterization of plasmids and other genetic elements is also epidemiologically useful

Methods: To characterize uropathogenic strains of E. coli, we studied 96 E. coli strains recovered from urine samples of children aged 1 month to 14 years with community-acquired UTIs in Jahrom, Iran. We assessed virulence factors [VFs], drug sensitivities, and plasmid profiles

Results: Drug sensitivities of the isolates were: 19.8% [ampicillin], 24% [trimethoprim-sulfamethoxazole], 29.2% [tetracycline], 75.5% [nalidixic acid], 80.4% [cefixime], 84.6% [gentamicin], 91.4% [ciproAoxacin], 96.8% [nitrofurantoin], 96.8% [amikacin] and 100% [imipenem]. Totally, 76 isolates harbored plasmids with an average of 5.5 plasmids [range: 1-10] in each strain. Plasmid profiling distinguished 22 different E. coli genotypes in all isolates that ranged in similarity from 50% to 100%. PCR showed that the prevalence of virulence genes ranged from 15.62% forhly to 30.2% for pap

Conclusion: These data mandate local monitoring of drug resistance and its consideration in empirical therapy of E. coli infections. Plasmid analysis of representative E. coli isolates also demonstrates the presence of a wide range of plasmid sizes, with no consistent relationship between plasmid profiles and resistance phenotypes. Plasmid profiles distinguished more strains than did the antimicrobial susceptibility pattern

Comparison of community and healthcare-associated MRSA in Iran

To characterize and compare the epidemiological and microbiological aspects of community and healthcare-associated MRSA [CA-MRSA, and HA-MRSA] cases in Iran, this prospective cohort study was conducted from January to December 2008 in seven hospitals. Staphylococci were isolated from 109 hospitalized patients. MRSA isolates were classified into HA-MRSA and CA-MRSA based on clinical features. Antibacterial susceptibility patterns of the isolates to eight antibiotics routinely used to treat infected patients were determined according to standard agar dilution methods. Staphylococcal Cassette Chromosome mec [SCCmec] type of isolates and their correlation with antimicrobial susceptibility patterns in CA and HC isolates were determined. Of 109 isolates, 15[13.7%] were community-associated and 94 [86.3%] were healthcare-associated MRSA. The most frequent SCCmec types in the studied hospitals were SCC mec type I [56.9%] and type II [22%]. Relatively sulfamethoxazole, clindamycin, rifampin, erythromycin, tetracycline and doxycycline were noticed. To our knowledge, this is the first time that the analysis of SCCmec type is carried out in Iran according to the clinical criteria. Difference in the prevalence of HC-MRSA and CA-MRSA based on the clinical and epidemiological features may indicate the need for revisiting the classification of MRSA. The high prevalence of multi-drug resistant MRSA could be as a result of the excessive use of antibiotics in the hospitals. Therefore, periodical assessment of antibacterial susceptibility patterns of the MRSA strains is warranted

Il-1 beta [+3953 C/T] and IL-8 [-251 A/T] gene polymorphisms in H.pylori mediated gastric disorders

Previous studies imply that IL-1 and IL-8 gene variations may play a crucial role in the genetic predisposition to different gastric disorders upon H. pylori infection. The aim of this study was to determine the potential association between the prevalence of certain polymorphic sites and the risk of gastric disorders in Iranian population. One hundred and forty three unrelated individuals with different gastric disorders and 374 normal individuals with no gastric disorders and with a negative serology test for H. pylori [control group] were studied for the association between IL-1 beta [+ 3953 C/T] and IL-8 [-251 A/T] gene polymorphisms and H. pylori - mediated gastritis and gastric ulcer. An analysis of genotype frequency for these genes was performed using RFLP- PCR. Based on the data obtained from culture and pathologic findings, the patients were classified into three subpopulations: H pylori [+] non-ulcerative gastritis [+], H. pylori [+] ulcerative gastritis [+] and H. pylori[-] non-ulcerative gastritis [+]. A significantly higher frequency of TT genotype [p=0.02] in IL-1 beta +3953 in H.pylor[+] ulcerative gastritis [+] was revealed compared to the control group. There were no significant differences among other subpopulations. No significant differences in allele and genotype frequencies of IL-8 [-251A/T] were found among the patients. The data suggest that TT genotype in IL- 1 beta +3953 may be a major contributing genetic risk factor for H. pylori induced gastric ulcer. Moreover, the role of other bacterial and host response factors, such as bacterial adherence peptides, host chemokines, and genes involved in gastric acid secretion, must be further investigated in different ethnic populations

Changing prevalence of Helicobacter pylori in south of Iran

The prevalence of Helicobacter pylori has declined rapidly in Asia. This has been shown in both seroprevalence-based and endoscopy-based studies. The present study was conducted to determine the prevalence of gastric infection caused byH. pylori in an Iranian population residing in south of Iran. A total of 522 patients [266 females and 256 males with the mean age of 44.3 +/- 13.0, range 18-83 years] underwent endoscopy in Shiraz, southern Iran. The diagnosis of H. pylori infection was established by rapid urease test, culture and gram staining and the gastric disease was confirmed by an expert pathologist. From ulcerative [n=296] and non-ulcerative [n=226] patients, 156 [52.7%] and 94 [41.6%] H. pylori strains were isolated by culture, respectively. The prevalence of H. pylori infection was significantly higher in patients aged 21-30 and >50 years [66.66% and 62.12%, respectively]. However, H. pylori was not detected in 22 patients aged < 20 years The present study revealed a significant decline in the prevalence of H pylori infection in the studied population. It seems that in parallel with better therapeutic approaches and elimination of bacteria, an improvement in the personal hygiene and living conditions of the Iranian population contribute to lower prevalence of H. pylori

Comparison of arbitrarily primed-polymerase chain reaction and plasmid profiles typing of Pseudomonas aeruginosa strains from burn patients and hospital environment

To identify the strengths and weakness of arbitrary primed-polymerase chain reaction [AP-PCR] and plasmid profiles for typing of Pseudomonas aeruginosa [P. aeruginosa] and tracking of source of infections. Seventy-four strains of P. aeruginosa were isolated from burn patients and hospital environment between January to April 2003 in Ghotbadden Burn Hospital, Shiraz, Iran. The strains were classified by photo Capt Mw program, similarity and clustering of strains were assessed using NTSYS-PC version 2.02K software. Based on 50% and 64.7% and 67.5% similarity on the plotted dendrogram, 38 plasmid profiles were classified into: 2, 3 and 5 clusters, respectively. Photo Capt Mw program categorized AP-PCR products to 47 different types of 6 to 12 bands between 0.376 to 3.7 kb. Based on dendrogram pattern 3 levels [62%, 81% and 84.6%] of similarity were selected. Using these criteria 2, 5and 11 clusters were obtained, respectively. As compared with plasmid profiles, AP-PCR analysis protocol is rapid, reproducible and differentiated the isolates with higher discrimination power. These results suggest that during admission of patients in burn center a limited number of common strains cross-contaminate burn victims. However, transmissions of infection from hospital environment to patients also occur in the minority of the victims. To control cross-contamination of the patient wounds with antibiotics resistant isolates, strong disinfection of patients' bathroom after scrubbing of each patient wounds is mandatory

Discrimination of dominant helicobacter pylori strains isolated from patients with different gastroduodenal pathologies by protein profiling

It is not clear what factors determine divergent outcomes of infections caused by H. pylori. In the present study, the protein profiles of different strains of H. pylori, isolated from three groups of patients with ulcerative disease, non-ulcerative gastritis and cancer disease, were analyzed using 1D-SDS-PAGE. The patterns of different H. pylori strains were highly divergent. About 30.76% [7 bands] of the 26 observed protein bands were common in all strains isolated from 3 groups of the patients. While the similarity for the strains inside each group were 75% [15 from 20], 76.47% [13 from 17] and 78.57% [11 from 14] for cancerous, ulcerative and nonulcerative group, respectively. Some of the observed bands were significantly specific for each group. Therefore, we speculated that some H. pylori strains might be more associated with a specific disease than others, giving the clustering of some, but not all, strains within each disease group. In conclusion, this study showed that protein profile can be a characteristic in discrimination of dominant strains in different gastric clinical status. Specific and dominant proteins of different strains isolated from three groups of patients under study were candidates for further exploration for laboratory tests, which analyze disease-specific H. pylori strains, and for diagnosis of the different diseases and outcomes associated with this widespread bacterium

Effect of cyclophosphamide on the course of candida albicans infection in normal and vaccimated mice

To evaluate the immunomodulating effect of cyclophosphamide [Cy] on the course of Candida albicans [C. albicans]. We performed this study in the Shiraz Medical School, Shiraz, Iran during April to November 2003. Five groups of 10 mice [vaccinated group] were immunized by 5 equal injections of 2x105, 2.5x105 and 3x105 of the organism intraperitoneally. Then, the group received Cy on day zero and was challenged with lethal doses of C. albicans [7.74x105 colony forming unit] on days zero, one, 3, 6 and 12 post-Cy injection. Another 5 equal groups of 10 mice [non-vaccinated group] received Cy on day zero and similar to vaccinated ones were challenged with lethal doses of the organism too. The control groups received just Cy on day zero and were sacrificed on days zero, one, 3, 6 and 12 days post-Cy injection. We performed the hemogram and the spleen and studied the renal tissues microscopically and macroscopically. In vaccinated group, we observed an increase in survival time and in spleen and renal weights were visible while in non-vaccinated ones, a significant decrease was also observed on days one and 3 and an increased on days 6 and 12 post-Cy injection. We observed atrophy and necrosis in the spleen while inflammation and necrosis were also observed in the kidneys on days one and 3. We noticed a significant hyperplasia in the white pulp on days 6 and 12 post-Cy injection. We conclude that hyperplasia in the white pulp of spleen and the increase in peripheral polymorphonuclears due to selective effects of Cy could effectively protect the animal against C. albicans infection

Immunodominant antigens of helicobacter pylori strains isolated from patients with different gastroduodenal diseases

To detect the immunogenic proteins in Helicobacter pylori [H. pylori] strains isolated from patients with different gastric diseases. We performed this study in the Clinical Microbiology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran, during July 2003 to September 2004. Total proteins of H. pylori strains isolated from the gastric biopsies of 3 groups of patients were separated by 1D-SDS-PAGE and then blotted with the sera of their respective hosts. In SDS-PAGE the members of each group showed high correlation according to similarity in their patterns, resulting in considering them in the same cluster. The patterns of immunoblots differed from that of Coomassie Brilliant Blue stained gels. The blotting method did not recognize some of the protein bands in the SDS-PAGE. Only the bands of 106 and 45 kDa from H. pylori strains isolated from patients with gastric cancer were significantly [p<0.05] recognized specifically with the sera of their respective patients, and the band of 13 kDa was recognized specifically [p<0.05] with the sera of nonulceric patients. With the exception of these bands, in the patterns of blotting of the sera from all patients no significant differences were observed. By using 1D blotting methods we could find 2 antigenic protein bands [106 and 45 kDa] for H. pylori strains isolated from cancerous patients, and one [13 kDa] for the strains isolated from nonulceric patients, which were specifically recognized with their respective host

Modified DNA extraction for rapid PCR detection of methicillin-resistant staphylococci

Nosocomial infection caused by methicillin-resistant staphylococci poses a serious problem in many countries. The aim of this study was to rapidly and reliably detect methicillin-resistant-staphylococci in order to suggest appropriate therapy. The presence or absence of the methicillin-resistance gene in 115 clinical isolates of Staphylococcus aureus and 50 isolates of Coagulase Negative Staphylococci [CNS] was examined by normal PCR. DNA extraction for PCR performance was then modified by omission of achromopeptadiase and proteinase K digestion, phenol/chloroform extraction and ethanol precipitation. All isolates with MIC>8 micri g/ml showed positive PCR. No differences in PCR detection have been observed when normal and modified DNA extractions have been performed. Our modified DNA extraction can quickly detect methicillin-resistant staphylococci by PCR. The advantage of rapid DNA extraction extends to both reduction of time and cost of PCR performance. This modified DNA extraction is suitable for different PCR detection, when staphylococci are the subject of DNA analysis

Distribution patterns of methicillin resistance genes [mecA] in staphylococcus arureus isolated from clinical specimens

There is a growing concern about the application of molecular methods in epidemiological studies of infectious diseases. The objective of this study was to determine the genetic stability of methicillin resistance genes in Staphylococcus aureus for the evaluation of resistance strain distribution. One hundred and fifteen S. aureus isolates from patients with staphylococcal infection were collected. The isolates were screened for methicillin resistance by minimum inhibitory concentration [MIC] examination. The stability of methcillin resistance genes was examined by physical curing and PCR screening methods. The results showed that methicillin resistance Staphylococcus aureus [MRSA] had risen up to 43% in Nemazi Hospital [Shiraz, Iran]. Indeed, the incidence of MRSA in our hospital was 10% during the last four years. The ability to lose [curability] of methicillin resistance genes [mecA] was examined by physical curing method in 49 isolates with MIC >/= 16 micro g ml -1. No sign of curability of mecA gene was observed where 500 colonies from each strain have been studied and exhibited by the same MIC values before and after curing test. Positive PCR results for isolates with MIC >/= 16 micro g ml -1 before and after curing experiment have been achieved. These data confirm the results of curing method, indicating that stable genetic determinants confer methicillin resistance. These results support the hypothesis that resistance isolates may be selected due to clonal selection under antibiotic pressure used in clinics rather than transmission of mobile genetic determinant. The high prevalence of MRSA immerged in our Hospital could be originated due to antibiotic pressure and poor control measures