Developing the Molecular Genetic Component of Microbiological Monitoring of Nosocomial Acute Enteric Viral Infections

Abstract

The aim of the investigation was improving the effectiveness of microbiological monitoring through optimization and development of the molecular genetic component based on comprehensive long-term assessment of hospital-acquired acute enteric infections of viral etiology.Materials and Methods. In the framework of epidemiological surveillance of nosocomial viral infections, syndrome-based diagnosis of acute enteric infection cases was implemented in a children's hospital. In fact, the patients with the signs of gastrointestinal dysfunctions unrelated to the underlying disease were identified and examined. DNAs (RNAs) of acute enteric infections of viral etiology were detected and differentiated through PCR-based diagnosis. G[P]-typing of rotaviruses was performed by RT-PCR method using various primers. Genotyping of enteric viruses by sequencing method was performed determining the relevant nucleotide sequences of rotavirus, norovirus and astrovirus cDNA segments using genetic analyzer Beckman Coulter. Nucleotide sequences of cDNA segments were analyzed using BLAST programs to identify closely related strains and an online service for automatic genotyping of noroviruses. Alignment of nucleotide sequences and phylogenetic analysis were performed using MEGA software. Sequences of genome segments obtained in this study have been represented in the international GenBank database.Results. There has been developed the molecular genetic component of microbiological monitoring of acute enteric infections of viral etiology, which involves not only diagnosing the enteric pathogens by PCR, but also subsequent genotyping as well as phylogenetic analysis to determine genetic characteristics of pathogens.