scholarly journals EVALUATION OF A RAPID SCREENING ASSAY FOR BACTERIAL IDENTIFICATION (DOT-ELISA) IN FECAL SAMPLES FROM CHILDREN

Author(s):  
Etelvina BOCCATTO ◽  
Sânia Alves dos SANTOS ◽  
Neusa Augusta Oliveira MAZIERI ◽  
Dahir Ramos de ANDRADE ◽  
Francisco Roque CARRAZZA ◽  
...  

With the objective of standardizing a Dot Enzyme-Linked Immunosorbent Assay (Dot-ELISA) to detect antigens of fecal bacterial enteropathogens, 250 children, aged under 36 months and of both sexes, were studied; of which 162 had acute gastroenteritis. The efficacy of a rapid screening assay for bacterial enteropathogens (enteropathogenic Escherichia coli "EPEC", enteroinvasive Escherichia coli "EIEC", Salmonella spp. and Shigella spp.) was evaluated. The fecal samples were also submitted to a traditional method of stool culture for comparison. The concordance index between the two techniques, calculated using the Kappa (k) index for the above mentioned bacterial strains was 0.8859, 0.9055, 0.7932 and 0.7829 respectively. These values express an almost perfect degree of concordance for the first two and substantial concordance for the latter two, thus enabling this technique to be applied in the early diagnosis of diarrhea in infants. With a view to increasing the sensitivity and specificity of this immunological test, a study was made of the antigenic preparations obtained from two types of treatment: 1) deproteinization by heating; 2) precipitation and concentration of the lipopolysaccharide antigen (LPS) using an ethanol-acetone solution, which was then heated in the presence of sodium EDTA

2021 ◽  
Vol 2021 ◽  
pp. 1-12
Author(s):  
Victorien Dougnon ◽  
Edna Hounsa ◽  
Eric Agbodjento ◽  
Lunga Paul Keilah ◽  
Brice Boris Legba ◽  
...  

Previous work stated that Khaya senegalensis, Anacardium ouest L., Pterocarpus erinaceus, Diospyros mespiliformis, Ocimum gratissimum, Manihot esculenta, Vernonia amygdalina Delile, and Daniellia oliveri have a great potential for the fight against infectious diarrhea. However, data on their antibacterial activity on strains of bacteria responsible for infectious diarrhea are not available. This study is aimed at elucidating the mechanism of action of the antibacterial effect of these plants on some bacterial strains responsible for diarrheal infections. The design of the study included first evaluating the degree of sensitivity of Salmonella typhimurium 14028, Escherichia coli ATCC 25922, Shigella spp., and Salmonella spp. strains to aqueous and hydroethanolic extracts of each plant, followed by the determination of minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC), and antibiotic power (Pa). This screening was completed with the evaluation of the possible mode of action of the extracts by testing the membrane permeability of these bacterial strains. The data collected indicate that the bacterial strains tested were sensitive to the extracts to varying degrees, except Cassia sieberiana DC and Pseudocedrela kotschyi extracts. For the active extracts, inhibition diameters ranged from 18.33 mm to 7 mm. With the exception of Escherichia coli, all strains were sensitive to the aqueous and hydroethanolic extracts of Anacardium occidentale. MICs vary between 3.37 and 25 mg/ml. Membrane permeability test data show that all active extracts affect the bacterial strains tested by attacking the stability of their outer membrane. For all active extracts, the high percentage of membrane destabilization of the bacteria is significantly ( p < 0.05 ) better than that of cefixime used as a reference. Thus, it appears that these extracts can destroy Gram-negative bacteria and increase the fluidity and permeability of their cytoplasmic membrane. The knowledge of the mechanism of action of these extracts is an interesting contribution to the fundamental knowledge on the alternative that medicinal plants represent to antibiotics. These extracts can be used in the management of infectious diarrhea.


2007 ◽  
Vol 70 (10) ◽  
pp. 2230-2234 ◽  
Author(s):  
T. W. THOMPSON ◽  
T. P. STEPHENS ◽  
G. H. LONERAGAN ◽  
M. F. MILLER ◽  
M. M. BRASHEARS

Rapid enzyme-linked immunosorbent assays (ELISAs) are approved for detection of Escherichia coli O157 in beef products. However, these kits have also been used in the industry to detect this pathogen on hides or in feces of cattle, although this use has not been validated. The objective of this study was to compare commercially available ELISAs (E. coli Now, Reveal, and VIP) with immunomagnetic separation along with selective media to detect E. coli O157 on hides, in feces, and in medium- and low-level-inoculated ground beef and carcasses (simulated by using briskets) samples. Naturally infected hide and fecal samples were subjected to both the immunomagnetic separation method and ELISAs for the detection of E. coli O157. Additionally, E. coli O157 inoculated and noninoculated ground beef and beef briskets were used to simulate meat and carcass samples. When comparing the detection results from the ELISAs (E. coli Now, Reveal, and VIP) to the immunomagnetic separation method, poor agreement was observed for fecal samples (kappa = 0.10, 0.02, and 0.03 for E. coli Now, Reveal, and VIP, respectively), and fair-to-moderate agreement was observed for hide samples (kappa = 0.30, 0.51, and 0.29 for E. coli Now, Reveal, and VIP, respectively). However, there was near-perfect agreement between the immunomagnetic separation method and ELISAs for ground beef (kappa = 1, 1, and 0.80 for E. coli Now, Reveal, and VIP, respectively) and brisket (kappa = 1, 1, and 1 for E. coli Now, Reveal, and VIP, respectively) samples. Assuming immunomagnetic separation is the best available method, these data suggest that the ELISAs are not useful in detecting E. coli O157 from hide or fecal samples. However, when ELISAs are used on ground beef and beef brisket samples they can be used with a high degree of confidence.


1999 ◽  
Vol 62 (4) ◽  
pp. 325-328 ◽  
Author(s):  
C. LITTLE ◽  
D. ROBERTS ◽  
E. YOUNGS ◽  
J. de LOUVOIS

A study of imported unprepared whole lettuces sampled from supermarkets, greengrocers, shops, and market stalls found that all were of acceptable microbiological quality. Twenty-seven out of 151 (18%) imported lettuce samples had Enterobacteriaceae levels of 104 CFU/g or more. However, these bacteria that constitute part of the natural microflora of unprepared vegetables may also be derived from the soil and/or by poor handling. The pathogens, Salmonella spp., Shigella spp., Campylobacter spp., Escherichia coli O157:H7, Vibrio cholerae, Listeria monocytogenes, and also Escherichia coli, an indicator of fecal contamination, were not detected in any imported lettuces, indicating that hygiene, harvesting, and production practices were good. Imported lettuces with Enterobacteriaceae levels of 104 CFU/g or more varied with type of retail premises and the temperature at which the lettuces were displayed. Samples from greengrocers, shops, and market stalls were more likely to contain Enterobacteriaceae at levels in excess of 104 CFU/g than those from supermarkets.


Pathogens ◽  
2020 ◽  
Vol 9 (9) ◽  
pp. 672
Author(s):  
Néstor Casillas-Vega ◽  
Fernanda Flores-Rodríguez ◽  
Israel Sotelo-Coronado ◽  
Magda Elizabeth Vera-García ◽  
Aldo García-Heredia ◽  
...  

Little information is available regarding the pathogens that cause diarrhea in hospitalized patients who also have various clinical problems. The purpose of this study was to determine the presence of pathogens in fecal samples of hospitalized patients all suffering diarrhea in addition to other problems in Mexico. Diarrheic stools from 240 patients were obtained in a third-level hospital in Monterrey, Mexico. PCR was used for the detection of Salmonella spp., Shigella spp., Campylobacter spp., Yersinia spp., Aeromonas spp., Clostridioides difficile, and norovirus GI and GII. The presence of trophozoites, cysts of protozoa, eggs, and/or helminth larvae was determined by microscopic observation. Of the 240 patients analyzed, 40.4% presented at least one of the pathogens analyzed. Norovirus was the pathogen most frequently found (28.6%), followed by bacteria (11.7%), and parasites (8.3%). The majority of co-infections were parasites + norovirus, and bacteria + norovirus. Norovirus was detected mainly in children aged 0 to 10 years (9/15, 60%). Patients aged 0–20 years did not present co-infections. Entamoeba coli and Entamoeba histolytica were the most common parasites, (8/240), and Salmonella was the most prevalent bacteria (10/240). This information can help design specific strategies useful for hospitalized people with a compromised status.


2017 ◽  
Vol 100 (4) ◽  
pp. 1038-1050 ◽  
Author(s):  
Brian Bullard ◽  
Curtis H Stumpf ◽  
Weidong Zhao ◽  
Stephanie Kuzenko ◽  
Gary D Niehaus

Abstract The Crystal Diagnostics (CDx) Xpress S Kit is a rapid-screening assay for Salmonella spp. in whole raw tomatoes, whole chicken carcasses, raw ground beef, raw beef trim, and whole liquid pasteurized eggs with citric acid when present at levels of 1 CFU/portion size. The Xpress S system comprises an automatic CDx Xpress Reader and a single-use CDx BioCassette that incorporates antibody-coupled microspheres and liquid crystal for the selective identification of the intended microbe. In internal evaluations, the CDx Xpress S Kit detected all 142 Salmonella strains tested, including non-enterica subspecies, and excluded all non-Salmonella species assayed. Method-developer studies, as well as a third-party evaluation, demonstrated that 15 h single-stage enrichment permits rapid detection equivalent to the U.S. Department of Agriculture and U.S. Food and Drug Administration reference methods. The results demonstrate that the CDx Xpress S Kit is one of the fastest, most sensitive, and most accurate methods for detecting Salmonella in food matrixes.


2021 ◽  
Vol 7 (2) ◽  
pp. 379-386
Author(s):  
M. Abdulai ◽  
Z. I. Abubabakari ◽  
S. J. Cobinna ◽  
D. Oduro

Investigations for pathogenic microbes on surfaces of objects have been attaining significant consideration from public health researchers for some time now. This is especially because of the risk of the spread of diseases of public health concern. Consequently, this study was conducted to provide information on bacterial contamination levels in taxis and motorised tricycles operating from the Tamale Main Taxi Rank. Forty samples from taxis and tricycle seats were collected and analysed. All samples contained Escherichia coli, Salmonella spp, and Shigella spp. Taxis contained significantly (p < 0.006) more bacteria than tricycles (mean bacterial loads of 20.35×106 CFU/ml and 2.80×10 6 CFU/ml respectively). Also, considering the materials with which the vehicle seats were made vinyl contained more bacteria (14.06×106 CFU/ml) than cloth (10.98×106 CFU/ml) and polythene (1.12×106 CFU/ml). Using the Kruskal-Wallis test for analysing bacteria loads across material types, it was found that there was a significant difference (p < 0.026) between bacteria numbers and material types. The bacteria isolates showed resistance to the antibiotics, ampicillin (10 μg), Ceftazidime (30 μg), Cefotaxime (30 μg), and Ciprofloxacin (5 μg). Taxis and tricycles in some parts of the Metropolis operate under poor hygienic conditions. It is recommended that routine cleaning and disinfection of vehicles as well as the practice of personal hygiene be highly encouraged to keep a safer and healthier urban population and environment. Keywords: Escherichia coli, Pathogenic, Salmonella spp, Shigella spp


2019 ◽  
Author(s):  
Hailing Chang ◽  
Jiayin Guo ◽  
Zhongqiu Wei ◽  
Zheng Huang ◽  
Chuning Wang ◽  
...  

Abstract Background Diarrhea is still a major cause of childhood morbidity and mortality worldwide. This hospital-based study aimed to monitor the consecutive epidemiological trend of etiology in children with acute diarrhea in Shanghai.Methods Outpatient children with diarrhea were prospectively enrolled within 7 days after onset of diarrheal symptoms during 2015-2018. Fresh stool samples were collected for testing enteropathogens. Enteric bacteria were identified and typed through culture and serotyping. Enteric viruses were identified through real-time PCR assay. hadResults Enteric pathogens were identified in 1572 (58.4%) of the 2692 enrolled children with acute diarrhea. Viruses were more frequently detected than bacteria (41.3% versus 25.0%), and co-infection with 2 or more pathogens was found in 13% of outpatients. Nontyphoidal Salmonella spp . (NTS) was the most common bacteria with 10.3% of isolate rate, followed by enteropathogenic Escherichia coli (EPEC) (6.5%), enteroaggregative Escherichia coli (EAEC) (6.2%), Campylobacter spp . (3.6%), enterotoxigenic Escherichia coli (ETEC) (1.1%), Shigella spp . (0.2%), and enterohemorrhagic Escherichia coli (EHEC) (0.1%). Rotavirus was the most common virus with 16.0% of detection rate, followed by norovirus (15.5%), adenovirus (7.2%), sapovirus (3.0%) and astrovirus (2.7%).Conclusions Infectious diarrhea remains the major cause of diarrhea in children in Shanghai. Rotavirus, norovirus and NTS were the major enteric pathogens responsible for diarrhea in Shanghainese children. Improving uptake of rotavirus vaccine and strengthening prevention of foodborne pathogens will be helpful to reduce the burden of diarrheal diseases in children in Shanghai.


2014 ◽  
Vol 6 (2) ◽  
pp. 45-53
Author(s):  
Dubravka Milanov ◽  
Dragan Fabijan ◽  
Bojana Prunić ◽  
Maja Velhner ◽  
Tamaš Petrović

Fecal samples originating from 15 Eurasian griffon vultures were collected during June 2012 in the territory of special nature reservation Uvac and examined for presence of enteric bacteria Escherichia coli and Salmonella spp. Salmonellas were isolated from five samples (33.3%) and serologically typed as Salmonella enterica subsp. enterica ser. Veneziana. E. coli was isolated from four samples (26.6%). Antimicrobial susceptibility testing revealed resistance to one and more antibiotics only in E. coli isolates.


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