| Abstract [eng] |
Detection of Different Microorganisms Causing Diarrhoea by Real-Time PCR Diarrhoeal disease is the second leading cause of mortality and morbidity in children younger than five years old. Gastrointestinal infections caused by organisms such as bacteria, viruses or protozoa. The principle diagnostic modalities have historically included culture, microscopy and antigen based tests. But these methods aren’t always specific and selective. A new molecular diagnostics have an emerging role in the diagnosis of infectious diseases. The aim of this study was to detect and analyze gastrointestinal pathogens from stool samples by real-time polymerase chain reaction. In order to carry out the aim of the study, these tasks were established: 1) to detect the etiologic agents of diarrhoea in children using Real-Time PCR; 2) to evaluate the frequency of organisms causing infectious diarrhea; 3) to compare molecular method with standard microbiological methods involved with direct antigen detection, bacterial culture and microscopy used in infectious diarrhoea diagnostics. The detection of microorganisms causing diarrhea was performed at Vilnius University Hospital Santaros Clinics Centre of Laboratory Medicine. The study involved the analysis of the 53 stool samples of pediatric patients with gastroenteritis using commercial multiplex RT-PCR (AllplexTM Gastrointestinal Panel Assays) covering 25 diarrhea – causing pathogens. The results of this researched showed that 11 infectious gastrointestinal microorganisms from fecal samples were detected by using RT-PCR. Viral infections included with Rotavirus A (25%), Norovirus (15%) and bacterial infections included with C. difficile (12%), Campylobacter spp. (12%), showed the highest prevalence of infections diarrhea in children of 0-17 years group. By using RT-PCR were detected more infections diarrheal pathogens and mixed gastrointestinal infections than conventional methods. Newer molecular tests have the capability to detect small amount of nucleic acid of microorganisms, enumerate multiple pathogens and define the relative amounts in a single stool sample. |
