Comparison of direct and indirect immunofluorescence assays for rapid detection of cytomegalovirus in shell-vial culture.
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Biomedical subjects
Publications and source records attributed to J Reina.
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We report the microbiological characteristics of two Rahnella aquatilis strains isolated in the faeces of two patients with acute gastroenteritis, one of whom is an AIDS patient. The biochemical behaviour was studied with different automated identification systems, and the few clinical cases to be found in the literature were reviewed. Of the nine strains isolated in clinical samples, two were obtained from blood cultures, two from respiratory samples, one from urine, one from a burn wound, one from a surgical wound, and two (our strains) from faeces. In almost all cases the patient presented an underlying condition facilitating infection by opportunistic microorganisms. The majority of strains are characterized by their resistance to ampicillin, cephalothin and cefoxitin. Due to the rarity of the isolation of R. aquatilis in human samples it is not yet possible to establish, with any degree of certainty, its true pathogenic capacity.
We prospectively evaluated the efficacy of two commercial rapid methods for antigenic detection, a dot-blot enzyme immunoassay (EIA-DB) (Directigen FluA, Becton-Dickinson, USA) and a direct immunofluorescence assay (DIF) (Monofluokit Influenza A, Diagnostics Pasteur, France), compared with the shell-vial culture in the MDCK line, incubated 2 to 3 days and stained with the monoclonal antibody clone IA-52, the diagnosis of lower respiratory tract caused by Influenza A virus (IA). In the study period the presence of IA virus was detected in 59 of the 377 samples analyzed (15.7%). Only the SVC method detected all positive samples (100% sensitivity), being used as a reference method for comparison with the other techniques). The EIA-DB technique detected 50 cases (84.7%) and the DIF only 35 (59.3%). In nine (15.2%) cases the diagnosis was obtained only with the SVC method. The results of the comparison of the EIA-DB technique with SVC were: sensitivity 84.7%, specificity 100%, positive predictive value 100%, and negative predictive value 97.2%. The DIF technique gave values of 59.3%, 100%, 100%, and 92.9%, respectively. A statistically significant difference was observed between the sensitivity of the EIA-DB and the DIF method (p = 0.0001). In view of the results we recommended the use of the EIA-DB as a screening method when infection by the IA is suspected. But to obtain the maximum diagnostic yield all samples would be inoculated in a shell vial culture with the MDCK cell line.
A case of acute diarrhoea caused by Aeromonas trota (formerly HG 13 group) in a Spanish child is reported. The strain was isolated in the faeces using the CIN agar (cefsulodin-irgasan-novobiocin) culture media. The strain was initially identified as A sobria by the commercial GNI card and API 20E biochemical systems. The strain was, however, VogesProskauer and sucrose negative, so complementary tests of cellobiose fermentation and gluconate oxidation were performed. These tests, together with the strain susceptibility to ampicillin (MIC 1 microgram/ml) and carbenicillin (MIC < 16 micrograms/ml) led to the final identification of A trota. The microbiological characteristics of this new species and the principal tests required for its identification are presented. The isolation, for the first time, of A trota in the Mediterranean area confirms the suspected worldwide distribution of this species.
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The aim was to evaluate the efficacy of the API-Campy system in the biochemical identification of 62 hippurate negative campylobacter strains isolated from the faeces. The strains were identified manually as 34 nalidixic acid susceptible C coli (NAS), 20 nalidixic acid resistant C coli (NAR), and eight C lari. The 34 strains of NAS C coli were identified as such by the API-Campy system. Of the 20 strains of NAR C coli, 15 (75%) were correctly identified by the commercial system. None of the five NAR C coli strains which were also erythromycin resistant was identified as such by the system. The eight C lari strains could not be identified by the API-Campy system because the bionumber obtained does not exist in the database of the computer system. The API-Campy system could be very useful for the identification of NAS C coli. However, failure to allow for a higher percentage of resistance to nalidixic acid in this species does not permit good identification of NAR strains. More important discrepancies are observed in C lari strains. In order to improve the identification of NAR C coli and C lari stains, it is advisable to include, or recommend as complementary, the indoxyl acetate hydrolysis test.
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BACKGROUND: To study the prevalence of virulence factors (adhesion, invasion, cytotoxicity and hemolytic activity) and to establish the presence of pathovars (virulence phenotype) in C. jejuni strains isolated in pediatric patients with inflammatory and secretory diarrhea and asymptomatic carriers. METHODS: We analyzed 95 strains of 48 patients with inflammatory diarrhea (blood and mucus in feces), 30 patients with secretory diarrhea (watery) and 17 strains isolated in asymptomatic children (control group). The study of adherence capacity, invasion and cytotoxicity was made in the Hep-2 cell line, and the analysis of hemolytic activity in blood agar plates with a 5% sheep's blood. The pathovars were defined by the cellular adhesion (phenotypes A and a) and the cytotoxicity (phenotypes E and e). RESULTS: 29.1% of inflammatory strains presented adherence capacity, 66.6% were invasive, 64.5% cytotoxic and 52.1% hemolytic. In the secretory strains the values were 70, 20, 10 and 6.6% respectively; in the control group the 11.7% presented adherence capacity and 5.8% were invasive. We obtained difference statistically significative for the secretory strains in the adherence capacity, and in inflammatory strains in the adherence capacity, cytotoxicity and hemolysis. The phenotype Ae predominate in the secretory strains, and the phenotype ae in the strains belonging to the control group. No pathovar predominates in the inflammatory strains. CONCLUSIONS: The analysis of virulence markers permit us to establish the pathogenic behaviour of the C. jejuni strains isolated in patients with diarrhea. The study of the adherence capacity and cytotoxicity (pathovars) would be used as a virulence markers and to predict the inflammatory or secretory nature of the diarrhea caused by C. jejuni strains.
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We report the in vitro antibiotic susceptibility of 1,220 strains belonging to the thermotolerant Campylobacter species, isolated from the feces of pediatric patients with diarrhea in the period from 1987 to 1993. The strains were identified as 1,148 C. jejuni isolates and 72 C. coli isolates. The overall results show that the strains showed drug resistance as follows: 51.8% to ampicillin, 4.4% to clindamycin, 2.6% to chloramphenicol, 21.2% to tetracycline, and 1% to gentamicin. Twenty-one strains (1.7%) displayed resistance to the combination of amoxicillin-clavulanic acid, and 3.2% of the strains were resistant to erythromycin (MIC of > or = 4 micrograms/ml), with a notable difference according to the species under consideration. While C. jejuni remained stable at 0.9 to 4% resistance to erythromycin, for C. coli the percentages detected ranged from 0 to 33%, with overall rates of 2.5 and 15.2% for the two species, respectively. Resistance to nalidixic acid (MIC of > or = 32 micrograms/ml) was found in 27.2% of the strains (27.8% for C. jejuni and 18% for C. coli), and resistance to ciprofloxacin (MIC of > or = 4 micrograms/ml) was found in 24.2% of the strains for C. jejuni and 15.2% for C. coli). Cross-resistance between nalidixic acid and ciprofloxacin was found in 89.1% of the strains (type 1 mutants), while 10.9% were resistant to nalidixic acid but susceptible to ciprofloxacin (type 2 mutants).
BACKGROUND: The differential clinical and epidemiological characteristics of pediatric patients (< 14 years) with gastroenteritis caused by rotavirus or adenovirus were studied. MATERIAL AND PATIENTS: A retrospective study was performed in patients with viral gastroenteritis diagnosed from January 1990 to December 1992. The detection of viral particles was carried out by the agglutination system with latex particles sensitized with antibodies versus rotavirus and adenovirus. Clinical and epidemiological characteristics of the patients were collected reviewing the medical records and emergency charts. RESULTS: Two hundred eighty-nine patients corresponding to 240 cases of infection by rotavirus (83.1%) and 48 (16.9%) by adenovirus, were analyzed. Diarrhea by rotavirus predominated in the winter months (59.7% of the cases from December to March). Nonetheless, those caused by adenovirus were seen throughout the year with a stable incidence being observed. Statistically significant differences were seen in both groups (p < 0.001) with rotavirus infection having a greater tendency to admission and greater incidence of vomiting, fever (> 39 degrees C) and underlying disease. The presence of leukocytosis was different in the adenovirus group as were the greater length of duration of diarrhea prior to attending the Emergency Department (5.2 days) and the total length of stay (11.2 days). No other associated enteropathogenic organisms were detected in 86.3% of the infections by rotavirus and in the 89.8% of those caused by adenovirus. In regard to age group distribution prevalence of diarrhea by rotavirus was observed during the first 3 months of life (76.6%) while adenovirus group it went up to 24 months of age with similar percentages. CONCLUSIONS: Diarrhea by rotavirus is mainly presented in the winter, with vomiting and its shorter in length and the symptoms less prolonged than diarrhea caused by adenovirus. Rotavirus diarrhea generally affect patients admitted with some underlying disease. Nonetheless, microbiologic diagnosis is required to establish the definitive etiology of both clinical processes.