Desarrollo de un método diagnóstico de tipo PCR múltiple para la detección de patógenos causantes de gastroenteritis aguda en niños mexicanos

Abstract

Acute gastroenteritis is a major cause of childhood morbidity and mortality worldwide. The etiology are included viruses, bacteria and parasites, considering the viruses the leading cause of acute diarrhea, rotavirus, calicivirus, enteric adenovirus and human astrovirus (HAstV) are the most frequent, in the bacterial etiology the main agents are Escherichia coli and Shigella spp. and in the etiology parasitic Entamoeba histolytica is recognized as the leading cause worldwide. The diagnosis of these pathogens is very little specific and sensitive, its conventionally used ELISA and molecular methods currently. However, actually don´t exists there is no a molecular method to identify them simultaneously, therefore the aim of this study was to design a molecular diagnostic method for highly sensitive and specific to allow the simultaneous identification in stool samples of enteric pathogens (astrovirus, norovirus, Entamoeba histolytica , enteroinvasive Escherichia coli) in children under four years. Stool samples were analyzed to identify these pathogens by ELISA and RT-PCR/PCR. For RT-PCR identified 4 samples positive for astrovirus (3.8%) and 4 for calicivirus (3.8%) were identified; while by PCR 2 and 14 samples positive for Entameoba histolytica (10.5%) and EIEC (70%) were identified respectively. By ELISA only one sample was positive for norovirus (2.5%). The sequences of the positive samples corresponded to serotype 1 and 8 for astrovirus, Norwalk virus, and EIEC strain. Multiplex PCR was standardized with positive stool samples, achieving simultaneously amplify (astrovirus, EIEC and Entamoeba histolytica), which is a useful tool in the identification of pathogens in epidemiological studies and may provide support to the diagnosis of acute gastroenteritis in children which would represent a major impact on early diagnosis and treatment and therefore a major epidemiological change.