Comparison of nested, multiplex, qPCR; FISH; SeptiFast and blood culture methods in detection and identification of bacteria and fungi in blood of patients with sepsis.

BACKGROUND: Microbiological diagnosis of sepsis relies primarily on blood culture data. This study compares four diagnostic methods, i.e. those developed by us: nested, multiplex, qPCR (qPCR) and FISH with commercial methods: SeptiFast (Roche) (SF) and BacT/ALERT® 3D blood culture system (bioMérieux). Blood samples were derived from adult patients with clinical symptoms of sepsis, according to SIRS criteria, hospitalized in the Intensive Care Unit. RESULTS: Using qPCR, FISH, SF, and culture, microbial presence was found in 71.8%, 29.6%, 25.3%, and 36.6% of samples, respectively. It was demonstrated that qPCR was significantly more likely to detect microorganisms than the remaining methods; qPCR confirmed the results obtained with the SF kit in all cases wherein bacteria were detected with simultaneous confirmation of Gram-typing. All data collected through the FISH method were corroborated by qPCR. CONCLUSIONS: The qPCR and FISH methods described in this study may constitute alternatives to blood culture and to the few existing commercial molecular assays since they enable the detection of the majority of microbial species, and the qPCR method allows their identification in a higher number of samples than the SF test. FISH made it possible to show the presence of microbes in a blood sample even before its culture.

[The frequency of anemia in children with newly diagnosed Crohn's disease in children]. / Czestosc wystepowania niedokrwistosci u dzieci z nowo rozpoznana choroba Lesniowskiego-Crohna.

UNLABELLED: The aim of the study was to analyze the occurrence of anemia in children with newly diagnosed Crohn's disease and 3 months after the initiation of treatment depending on location and activity of the disease. MATERIALS AND METHODS: 36 children aged from 2 to 18 years with newly diagnosed Crohn's disease treated in the years 2005-2011. According to the classification of Paris children were divided into 2 groups: II - 9 children (0-10 years); II - 27 children (11-17 years). We analyzed RBC, Hb, HCT, MCV, Fe, PCDAI at diagnosis and 3 months after the treatment, depending on the location and phenotype of the disease. Anemia was defined according to WHO criteria. Children were treated in accordance with the standards and activity of the disease. During the observation period supplementation of iron was not used. We analyzed the potential factors that could affect on the incidence of anemia. RESULTS: In the group I mean activity of the disease was lower than in group II in both observed periods. No statistically significant differences in hemoglobin, red blood cell count, hematocrit and volume of erythrocytes at diagnosis and after 3 months were noted. The average iron concentration increased from 6.98 microg/dl to 9.95 microg/dl (p < 0.05) after the treatment. A statistically significant higher incidence of iron deficit in group II in both analyzed periods was observed, respectively, 66.7% vs 92.59% and 55.5% vs 77.77% (p < 0.05). Anemia was more frequent in group II (p > 0.05). After 3 months of treatment the incidence of anemia reduced from 63.88% to 44.4% (p < 0.05) and iron deficit from 83.3% to 72.2% (p > 0.05). Inflammatory changes in upper gastrointestinal tract were found in 61.1% of children with Helicobacter pylori infection in 8.3% of cases. Ileal location was observed in 52.7% of children, more often in group II than I, 62.9% vs 22.22% (p < 0.05). Isolated location of inflammatory lesions in the colon more frequently observed in group I than II, respectively 55.5% vs 29.6% (p < 0.05). Thiopurine were used in 38.8% of children more often in group II than I, respectively 48.1% vs 11.1% (p < 0.05). Corticosteroids were used in 50.0% of children, often in group II than I, respectively 55.5% vs. 33.3% (p > 0.05). CONCLUSIONS: In children with newly diagnosed Crohn's disease anemia was diagnosed in 63.88% of the children and in 47.2% of children after 3 months of treatment (p < 0.05), iron deficiencies, respectively, in 83.3% and 72.2% of children (p > 0.05). 3-month treatment period of Crohn's disease has increased the iron concentration from 6.98 microg/dl to 9.95 mg/dl although the absence of supplementation (p < 0.05). In group II more often than in group I risk factors of anemia like changes in the upper gastrointestinal tract, the location of the ileum, intestinal villous atrophy and the use of immunosuppressive drugs were observed (p < 0.05). Anemia and iron deficiency were more often observed in the ileocolonic location than when the upper gastrointestinal tract was involved (adquately 55.5% vs 30.5%; p < 0.05 and 75.0% vs 41.6%; p < 0.05).

[Frequency of anemia in ulcerative colitis in children]. / Ocena czestosci wystepowania niedokrwistosci we wrzodziejacym zapaleniu jelita grubego u dzieci.

UNLABELLED: Anemia is one of most frequent complication of ulcerative colitis. It affects patients in the exacerbation of the disease as well as in the remission. It may be caused by the loss of the blood, the disturbances of intestinal absorption or deficient diet. The aim of the study was to estimate the rate of anemia in children with ulcerative colitis during first hospitalization. MATERIAL AND METHODS: The study comprised 36 children aged 2.5 to 18 years, 23 girls and 13 boys in whom ulcerative colitis was diagnosed based on the Porto criteria. Anemia in children was assessed depending on the age, period between first symptoms and diagnosis of the disease and on the activity of the disease according to PUCAI scale. RESULTS: Anemia was diagnosed in 72.2% of the children. Hemoglobin concentration ranged from 6.3 to 12.4 g/dl, average 10.05 g/dl depending on age. Activity of the disease according to PUCAI scale amounted to 5 to 80 points, average 51.9 points. Time from first symptoms to the diagnosis of the disease extended from one week to 12 months, average 3.6 months. In children without anemia the average concentration of hemoglobin was 13.2 g/dl, the activity of the disease 33.5 points and average time before diagnosis - 1.7 months. CONCLUSIONS: Anemia is a frequent symptom of ulcerative colitis in children. The presence of anemia depends on the disease activity and on the time between first symptoms and diagnosis of the disease. There is a necessity of iron supplementation in children with anemia despite effective anti-inflammatory treatment.