Intestinal nosocomial infections remain a major concern in paediatric wards leading to increased morbidity and mortality. This study determined the aetiological and epidemiological profile of nosocomial diarrhoea (ND) among children admitted to a hospital in India. During the period of January 2008 to June 2009, we consecutively enrolled 100 children between the age of 2 months and 14 years who developed ND as defined by the Centers for Disease Control and Prevention. A control group of patients matched for age and severity score but with no diarrhoea at admission or during their hospital stay (n550) were also enrolled. Stool samples were cultured for various pathogens using standard protocols. Clostridium difficile toxins and rotavirus antigen were detected using commercial ELISAs. Detection of diarrhoeagenic Escherichia coli was carried out by multiplex PCR assay. All patient details were noted. In this study, males predominated (77 %), and 56 % children were ,1 year of age and 96 % were ,5 years. The mean duration of diarrhoea and hospitalization in the case group was 3.2 days and 27.5 days, respectively. Malignancy and nasogastric tube usage were significant underlying factors for the development of ND. Diarrhoeagenic E. coli was the commonest agent (47 %: enterotoxigenic E. coli, enteroaggregative E. coli and enteropathogenic E. coli were isolated in 22, 18 and 7 % of patients, respectively). C. difficile toxin was seen in 9 % of cases, whilst rotavirus was found in 8 % of cases. Although rotavirus and C. difficile are major causative agents of hospital-acquired diarrhoea in the developed world, in this setting diarrhoeagenic E. coli was responsible for the majority of cases of hospital-acquired diarrhoea. ND was most common in children aged ,5 years.

BackgroundWe evaluated a dipstick test for rapid detection of Shigella sonnei on bacterial colonies, directly on stools and from rectal swabs because in actual field situations, most pathologic specimens for diagnosis correspond to stool samples or rectal swabs.Methodology/Principal FindingsThe test is based on the detection of S. sonnei lipopolysaccharide (LPS) O-side chains using phase I-specific monoclonal antibodies coupled to gold particles, and displayed on a one-step immunochromatographic dipstick. A concentration as low as 5 ng/ml of LPS was detected in distilled water and in reconstituted stools in 6 minutes. This is the optimal time for lecture to avoid errors of interpretation. In distilled water and in reconstituted stools, an unequivocal positive reaction was obtained with 4 x 106 CFU/ml of S. sonnei. The specificity was 100% when tested with a battery of Shigella and different unrelated strains. When tested on 342 rectal swabs in Chile, specificity (281/295) was 95.3% (95% CI: 92.9% - 97.7%) and sensitivity (47/47) was 100%. Stool cultures and the immunochromatographic test showed concordant results in 95.5 % of cases (328/342) in comparative studies. Positive and negative predictive values were 77% (95% CI: 65% - 86.5%) and 100% respectively. When tested on 219 stools in Chile, Vietnam, India and France, specificity (190/198) was 96% (95% CI 92%–98%) and sensitivity (21/21) was 100%. Stool cultures and the immunochromatographic test showed concordant results in 96.3 % of cases (211/219) in comparative studies. Positive and negative predictive values were 72.4% (95% CI 56.1%–88.6%) and 100 %, respectively.ConclusionThis one-step dipstick test performed well for diagnosis of S. sonnei both on stools and on rectal swabs. These data confirm a preliminary study done in Chile.

Ordered libraries facilitate large genetic screens by guaranteeing high genomic coverage with a minimal number of mutants, and they can save time and effort by reducing the need to construct targeted mutations. This resource is now available for P. mirabilis , a common and complicating agent of catheter-associated urinary tract infection.

Shigellosis or bacillary dysentery is an important cause of diarrhea, with the majority of the cases occurring in developing countries. Considering the high disease burden, increasing antibiotic resistance, serotype-specific immunity and the post-infectious sequelae associated with shigellosis, there is a pressing need of an effective vaccine against multiple serotypes of the pathogen. In the present study, we used bio-informatics approach to identify antigens shared among multiple serotypes of Shigella spp. This approach led to the identification of many immunogenic peptides. The five most promising peptides based on MHC binding efficiency were a putative lipoprotein (EL PGI I), a putative heat shock protein (EL PGI II), Spa32 (EL PGI III), IcsB (EL PGI IV) and a hypothetical protein (EL PGI V). These peptides were synthesized and the immunogenicity was evaluated in BALB/c mice by ELISA and cytokine assays. The putative heat shock protein (HSP) and the hypothetical protein elicited good humoral response, whereas putative lipoprotein, Spa32 and IcsB elicited good T-cell response as revealed by increased IFN-γ and TNF-α cytokine levels. The patient sera from confirmed cases of shigellosis were also evaluated for the presence of peptide specific antibodies with significant IgG and IgA antibodies against the HSP and the hypothetical protein, bestowing them as potential future vaccine candidates. The antigens reported in this study are novel and have not been tested as vaccine candidates against Shigella. This study offers time and cost-effective way of identifying unprecedented immunogenic antigens to be used as potential vaccine candidates. Moreover, this approach should easily be extendable to find new potential vaccine candidates for other pathogenic bacteria.

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