CLOSTRIDIUM DIFFICILE INFECTION IN CHILDREN WITH EXPOSURE TO ANTIBIOTICS.

Clostridium difficile represents an example of healthcare-associated pathogen causing disease in community and hospital setting. The number of clinical studies determining risk factors for Clostridium difficile infection in children is limited as compared to adult patients. The aim of this study was to determine the association between antibiotic exposure in hospital setting and risk of Clostridium difficile infection in children. Systematic review of the electronic databases was used to update the literature, collect secondary data, critically appraise research studies, and synthesize study result from our institutions with available literature. Paper aimed to provide a complete, exhaustive summary of current literature relevant to our research question. A systematic review of the literature using databases PubMed (1935-2018) was conducted to compare our study results with available literature and determine significant differences and similarities that might impact clinical decisions in the country Georgia. The study of the association of several factors with the development of Clostridium difficile infection showed that the age more than 4 years old represented independent risk-factors of the development of Clostridium difficile infection, while the gender and ethnicity was not associated with the infection. Antibiotic exposure within the month preceding the onset of diarrhea was statistically significantly associated with an increased risk of Clostridium difficile infection. The risk was greatest with cephalosporins, followed by penicillins, carbapenems and macrolides. Antibiotic exposure in children is associated with increased risk of Clostridium difficile infections in hospital setting.

EPIDEMIOLOGY OF CLOSTRIDIUM DIFFICILE INFECTION IN HOSPITALISED PEDIATRIC PATIENTS IN GEORGIA.

Clostridium difficile, a Gram-positive spore-forming bacillus, is the most common identifiable etiologic agent of antibiotic-associated diarrhea. The incidence of Clostridium difficile infections among hospitalized children has been increasing across the world. The aim of our study was to evaluate occurrence of Clostridium difficile and some other gastrointestinal pathogens among hospitalized pediatric patients in Georgia, as far as currently statistical data on the topic is very limited in the country. One of the objectives of the study was to test and pilot the real-time Polymerase Chain Reaction diagnostic systems for rapid and simultaneous identification of number of pathogens with a particular emphasis on diarrheal disease diagnostics as these are one of the primary public health priorities in Georgia and worldwide. Cross-Sectional study has been performed on 211 samples collected from 192 pediatric patients. Two pediatric hospitals were involved in the study: M. Iashvili Children's Central Hospital and Tbilisi Children's Clinical Hospital for Infectious Diseases. Laboratory investigations were done in the Clinic NeoLab, Tbilisi, Georgia. Study materials collected for testing were stool samples. Samples were tested by EIA kits (CerTest biotec, Zaragoza, Spain) for presence of A/B toxin according to the manufacturer's instructions. EIA test positive samples were analyzed by home-made multiplex real-time polymerase chain reaction (NeoPCR Diagnostics, NeoLab, Tbilisi, Gerogia) for confirmation of the infection and for simultaneous identification of additional gastrointestinal pathogens including Entamoeba histolitica, Giardia lamblia, Cryptsporidium parvum, Adenovirus, Rotavirus, Norovirus and Astrovirus. All samples were also tested for the presence of the above listed pathogenic agents using the same type EIA kits as for Clostridium difficile described above (CerTestbiotec, Zaragoza, Spain) for presence of the corresponding pathogen. The average age of the study participants was 3.5 years, 56.7% were male and 43.3% were female patients. Presence of Clostridium difficile have been documented in 21 samples out of 211 (10%). Besides the Clostridium difficile, other enteric pathogens have been revealed with the following frequencies: Rotavirus in 12 cases (5.7%), Adenovirus in 11 (5.2%), Giardia lamblia in 10 (4.7%), Astrovirus in 3 (1.4%), Cryptsporidium parvum in 3 (1.4%), Entamoeba histolitica in 2 (0.9%), Norovirus in 2 (0.9%). 49 samples were from out-patient cases (2 samples were positive for Clostridium difficile) and 162 samples were from in-patient cases (19 samples were positive for Clostridium difficile). Clostridium difficile is the frequent pathogenic agent causing diarrheal disease among hospitalized pediatric patients. Development of Clostridium difficile related diarrhea is associated with the antibiotic treatment of pediatric patients hospitalized due to different clinical diagnosis. Targeted early detection of these pathogens is important for the optimal management of diarrheal infection in pediatric patient which will lead to the better clinical outcome and reduction of morbidity rate among hospitalized pediatric patients.

RAPID IDENTIFICATION OF THE ETIOLOGICAL FACTORS CAUSING DIARRHEAL DISEASES.

The main objective of this investigation was to develop and pilot a real-time Polymerase Chain Reaction (rt-PCR) diagnostic system for rapid and simultaneous identification of pathogens with a particular emphasis on diarrheal disease diagnostics. The diarrheal diseases were selected as a target for the pilot because they constitute a primary public health priority in Georgia and worldwide. The product developed by our research team "Neo_PCR_Diagnostics" represents an original system for the identification of pathogens associated with gastrointestinal tract infections and diarrhea. The advantages of the proposed technology over existing conventional methods include the ability of simultaneous identification of multiple pathogens and the detection of pathogenic agents directly from the fecal samples. For the evaluation of the new diagnostic system, stool samples were collected at collaborating hospitals and clinics and analyzed by real-time PCR using the Neo_PCR_Diagnostic system. The selection of the pathogens for detection was based on their epidemiological and clinical importance. The following bacterial pathogens were targets for detection: Salmonella spp., Campylobacter spp., Shigella spp., Clostridium difficile (Toxin A/B), Escherichia coli (ETEC, STEC and O157), Yersinia enterocolitica and Vibrio cholerae. The following viral pathogens were studied: adenoviruses, rotaviruses and noroviruses. Genetic material (DNA) of the following parasites were targets in our study: Giardia lamblia, Entamoeba histolitica and Cryptosporidium spp. We also compared the results obtained by our molecular technology with the conventional methods - bacterial culture (for bacterial growth) and ELISA (for bacterial toxins). For viral and parasitic pathogens, comparison tests were performed with immunochromatographic assays for direct detection of antigens in the stool samples or with the data obtained by use of home-made end-point PCR (where available). Advantages of the proposed technology over existing conventional technologies include the ability of simultaneous identification of diarrheal infections by multiple pathogens. The proposed test system allows the detection of pathogenic agents directly from the fecal samples and can be completed within one working day. In general, the spectrum of pathogens detected by our approach was wider than those detected by the conventional methods used in the clinical setting, taking into consideration the list of pathogenic agents requisitioned by physicians within the framework of the routine clinical visit. Given these promising results, Neo_PCR_Diagnostics test performance and accuracy may be sufficient for use in molecular microbiological diagnostics in clinical and/or research settings.