Development of a multiplex pcr assay to detect gastroenteric pathogens in the feces of Mexican children

R. Tolentino-Ruiz, D. Montoya-Varela, M. García-Espitia, M. Salas-Benito, A. Gutiérrez-Escolano, C. Gómez-García, P. Figueroa-Arredondo, J. Salas-Benito, M. De Nova-Ocampo

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3 Citations (Scopus)

Abstract

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. © 2012 Springer Science+Business Media, LLC.
Original languageAmerican English
Pages (from-to)361-368
Number of pages324
JournalCurrent Microbiology
DOIs
StatePublished - 1 Oct 2012

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Feces
Mamastrovirus
Polymerase Chain Reaction
Entamoeba histolytica
Multiplex Polymerase Chain Reaction
Gastroenteritis
Escherichia coli
Enzyme-Linked Immunosorbent Assay
Parasites
Viruses
Morbidity
Bacteria
Mortality

Cite this

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. In: Current Microbiology. 2012 ; pp. 361-368.
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abstract = "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. {\circledC} 2012 Springer Science+Business Media, LLC.",
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Development of a multiplex pcr assay to detect gastroenteric pathogens in the feces of Mexican children. / 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.

In: Current Microbiology, 01.10.2012, p. 361-368.

Research output: Contribution to journalArticle

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Tolentino-Ruiz R, Montoya-Varela D, García-Espitia M, Salas-Benito M, Gutiérrez-Escolano A, Gómez-García C et al. Development of a multiplex pcr assay to detect gastroenteric pathogens in the feces of Mexican children. Current Microbiology. 2012 Oct 1;361-368. https://doi.org/10.1007/s00284-012-0167-7