Article
Development of a multiplex PCR assay to detect gastroenteric pathogens in the feces of mexican children
Fecha
2012-10Registro en:
Tolentino-Ruiz R, Montoya-Varela D, García-Espitia M, Salas-Benito M, Gutiérrez-Escolano A, Gómez-García C, Figueroa-Arredondo P, Salas-Benito J, De Nova-Ocampo M. Development of a multiplex PCR assay to detect gastroenteric pathogens in the feces of mexican children. 2012;65(4):361-8
Autor
Tolentino-Ruíz, Rocío
Montoya-Varela, Diana
García-Espitia, Matilde
Salas-Benito, Mariana
Gutiérrez-Escolano, Ana Lorena
Gómez-García, María del Consuelo
Figueroa-Arredondo, Paula
Salas-Benito, Juan Santiago
De Nova-Ocampo, Mónica
Institución
Resumen
Acute gastroenteritis (AGE) is a major cause of childhood morbidity and mortality worldwide; the etiology of AGE includes viruses, bacteria, and parasites.Amultiplex
PCR assay to simultaneously identify human Astrovirus (HAstV), Calicivirus (HuCVs), Entamoeba histolytica (E. histolytica), and enteroinvasive Escherichia coli (EIEC) in stool samples is described. A total of 103 samples were individually analyzed by ELISA (enzyme-linked immunosorbent assays) and RT-PCR/PCR. HAstV and HuCVs were
detected in four out of 103 samples (3.8 %) by RT-PCR, but ELISAs found only one sample as positive for HuCVs (2.5 %). E. histolytica was identified in two out of 19 samples (10.5 %) and EIEC in 13 out of 20 samples (70 %) by PCR, and all PCR products were sequenced to verify their identities. Our multiplex PCR results demonstrate the simultaneous amplification of different pathogens such as HAstV,
EIEC, and E. histolytica in the same reaction, though the HuCVs signal was weak in every replicate. Regardless, this multiplex PCR protocol represents a novel tool for the identification of distinct pathogens and may provide support for the diagnosis of AGE in children.