B. Transverse scan showing target sign appearance with the appearance of the appendicolith with its characteristic posterior acoustic shadowing. Collected data were statistically analyzed using χ 2 test. Continuous variables were analyzed using student’s t-test. P≤0.5 were considered statistically significant. Sensitivity and specificity were calculated for the CPGS. Kappa test was used to verify the specificity. All calculations were performed using

SAS version 8.2. Results In the current studied group of patients; age and sex analysis shows that cases with and without appendectomy are similar and there is no aggregation of cases in a certain age group or in a certain sex (Table 3). In 187 patients

(70.6%) appendectomy was performed, out of them 90 patients (48.1%) showed MCPGS between 15 and 22, those patients were kept with no oral feeding (NPO), intravenous fluid infusion SBI-0206965 mouse (IV fluid) of appropriate type and amount according to patient’s age before undergoing appendectomy. Only 8 out of the total appendectomies (4.3%) were normal at histopathological Belnacasan in vivo evaluation. The remaining 97 patients (36.6%) initially showed MCPGS of 8-14. On repeated evaluation every 2 hours for a maximum of 6 times and repetition of THI- US during the repeated evaluation for at least one time, their score progressed to 15 or more [61 patients (62.9%) with a MCPGS of 15-17, 11 patients (11.3%) with MCPGS

of 18, and 25 patients (25.8%) with MCPGS of 19]. During the observation period, no antibiotics were given in order not to alter the clinical picture. However, antibiotics were started once the diagnosis was confirmed. No false negative cases were recorded when using MCPGS. (Tables 3, 4) Table 3 Characteristics of studied children with clinically suspected appendicitis Character Number (%) oxyclozanide Age (months)      Minimum-maximum (mean ± SD) 18-203 (140.63 ± 25.923) Gender      Male 159 (60.0%)    Female 106 (40.0%) Referring site      None (parent decision) 229 (86.4%)    Health establishment (Pediatrician) 36 (13.6%) Duration of symptoms before admission (hours)      Minimum-maximum (mean ± SD) 6-48 (23.15 ± 11.182) MCPGS*      Minimum-maximum (mean ± SD) 1-22 (11.54 ± 6.113) Final Outcome      No surgery 78 (29.4%)    Appendectomy with negative histopathology 8 (3.0%)    Appendectomy with positive histopathology 179 (67.6%) MCPGS = Modified Clinical Practice Guideline Score Table 4 Comparing characteristics of children with and without appendicitis Character With Appendicitis# (n = 179) Without Appendicitis (n = 86) Test (P) Age (mean ± SD) 141.87 ± 23.584 138.06 ± 30.206 t = 1.12 (0.264) Gender     X2 = 0.413 (0.520)    Male 105 (58.7) 54 (62.8)      Female 74 (41.3) 32 (37.2)   Referring Agent     X2 = 0.015 (0.903)    None 155 (86.6) 74 (86.0)      Pediatrician 24 (13.4) 12 (14.

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With DNA from S. Enteritidis strains, the prot6E-specific, TET-labelled molecular beacons hybridise to their target amplicons and produce an orange fluorescent signal, whereas the fliC-specific, HEX-labelled molecular beacons remain dark. With DNA

from other Salmonella serotypes, no target amplicons are detected and both molecular beacons remain dark. The dashed line on the plots Selleckchem AZ 628 represents the normalised threshold for detection of fluorescence, the baseline above which fluorescence increases significantly on amplification and detection of the target sequence. In both the uniplex and double duplex assays, non-template controls were included to verify the absence of false-positive results. In all cases they exhibited undetectable amplification of the targets (CT >45). Selectivity of the real-time assay The selectivity and accuracy of the test is measured by calculating the values for specifiCity

and sensitivity. SpecifiCity is the probability that the PCR will be negative among specimens that should not possess the gene and is calculated using the formula: true negative/(true negative + false positive). Sensitivity shows the strength of the test in recognising what we are looking for, i.e., in correctly identifying the specific serotype. The formula used for estimation of sensitivity is: true positive/(true positive + false negative). For the reaction targeting the invA gene, all 44 Salmonella samples investigated were positive SBI-0206965 cost indicating a sensitivity of 100%. The specifiCity was also 100% since all non-Salmonella samples gave negative results, Calpain with undetectable fluorescence signals after 50 cycles. In the prot6E reaction all S. Enteritidis samples analysed were identified

correctly with positive PCR results and all non-Enteritidis samples were negative for this target. Thus, this reaction also had sensitivity and specifiCity of 100%. Similarly, in the assay for fliC detection, all S. Typhimurium samples tested were positive for the target. The assay’s sensitivity was 100%, matched by an equal specifiCity value as all non-Typhimurium samples tested gave negative PCR results. Discussion Traditional serotyping of S. enterica is based on the detection of certain antigens using microbiological techniques and culturing, which are time-consuming and laborious. This study exploits real-time PCR, molecular beacons and genetic variation between different serotypes to devise a quick, accurate and simple assay to reliably identify a bacterial sample as Salmonella enterica and further distinguish it as serotypes S. Typhimurium or S. Enteritidis, the two serovars most commonly associated with food-borne gastroenteritis. The assay described in this study can analyse a large number of samples very quickly, and can also identify as few as 10 copies of target DNA per reaction, potentially even in the presence of thousands of copies of other serotypes.

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