TY - JOUR
T1 - Development of a multiplex pcr assay to detect gastroenteric pathogens in the feces of Mexican children
AU - Tolentino-Ruiz, R.
AU - Montoya-Varela, D.
AU - García-Espitia, M.
AU - Salas-Benito, M.
AU - Gutiérrez-Escolano, A.
AU - Gómez-García, C.
AU - Figueroa-Arredondo, P.
AU - Salas-Benito, J.
AU - De Nova-Ocampo, M.
N1 - Funding Information:
This work was supported by the Instituto de Ciencia y Tecnología-DF and the Instituto Politécnico Nacional, (ICyT-DF/IPN-183).Tolentino-Ruiz,Rocío had a scholarship from CONACyT (270554/219933) and PIFI-IPN (SIP-20090206 y SIP-20100054); Montoya-Varela, D had a scholarship from ICyT-DF/IPN 183 project. The positive controls for all pathogens were gifts from Dr. E. Mendez, IBT UNAM, (fecal sample of Astrovirus); Dr. Carlos Eslava Campos, Departamento de Salud Pública Facultad de Medicina, UNAM (bacterial reference strain); and QBP Beatriz Alvarez Lara (positive samples of E. histolytica), Hospital General de México (SSA). DNA sequencing was performed by Alejandro Monsalvo Reyes (FES Iztacala-UNAM) and Dr. Laura Ongay, BSc Guadalupe Codiz (Instituto Fisiología Celular-UNAM). Dr. Gómez-García, C and Dr. Salas-Benito, J have fellowships from COFAA and EDI (IPN), and Dr. Figueroa-Arredondo, P and Dr. De Nova-Ocampo, M. have fellowships from COFAA (IPN).
PY - 2012/10
Y1 - 2012/10
N2 - Acute gastroenteritis (AGE) is a major cause of childhood morbidity and mortality worldwide; the etiology ofAGEincludes 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.
AB - Acute gastroenteritis (AGE) is a major cause of childhood morbidity and mortality worldwide; the etiology ofAGEincludes 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.
UR - http://www.scopus.com/inward/record.url?scp=84865991816&partnerID=8YFLogxK
U2 - 10.1007/s00284-012-0167-7
DO - 10.1007/s00284-012-0167-7
M3 - Artículo
C2 - 22711331
SN - 0343-8651
VL - 65
SP - 361
EP - 368
JO - Current Microbiology
JF - Current Microbiology
IS - 4
ER -