Development of a multiplex PCR assay to detect gastroenteric pathogens in the feces of Mexican children. 2012

R Tolentino-Ruiz, and D Montoya-Varela, and M García-Espitia, and M Salas-Benito, and A Gutiérrez-Escolano, and C Gómez-García, and P Figueroa-Arredondo, and J Salas-Benito, and M De Nova-Ocampo
Programa Institucional de Biomedicina Molecular, Sección de Estudios de Posgrado e Investigación, Escuela Nacional de Medicina y Homeopatía-IPN, Instituto Politécnico Nacional Guillermo Massieu Helguera No. 239, Col. Fracc La Escalera-Ticomán, 07320 México, DF, México.

Acute gastroenteritis (AGE) is a major cause of childhood morbidity and mortality worldwide; the etiology of AGE includes viruses, bacteria, and parasites. A multiplex 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.

UI MeSH Term Description Entries
D008800 Mexico A country in NORTH AMERICA, bordering the Caribbean Sea and the Gulf of Mexico, between BELIZE and the UNITED STATES, and bordering the North Pacific Ocean, between Guatemala and the UNITED STATES.
D002139 Caliciviridae A family of RNA viruses infecting a broad range of animals. Most individual species are restricted to their natural hosts. They possess a characteristic six-pointed starlike shape whose surfaces have cup-shaped (chalice) indentions. Transmission is by contaminated food, water, fomites, and occasionally aerosolization of secretions. Genera include LAGOVIRUS; NORWALK-LIKE VIRUSES; SAPPORO-LIKE VIRUSES; and VESIVIRUS. Nebovirus,Neboviruses
D002648 Child A person 6 to 12 years of age. An individual 2 to 5 years old is CHILD, PRESCHOOL. Children
D004748 Entamoeba histolytica A species of parasitic protozoa causing ENTAMOEBIASIS and amebic dysentery (DYSENTERY, AMEBIC). Characteristics include a single nucleus containing a small central karyosome and peripheral chromatin that is finely and regularly beaded. Endamoeba histolytica,histolytica, Endamoeba
D004926 Escherichia coli A species of gram-negative, facultatively anaerobic, rod-shaped bacteria (GRAM-NEGATIVE FACULTATIVELY ANAEROBIC RODS) commonly found in the lower part of the intestine of warm-blooded animals. It is usually nonpathogenic, but some strains are known to produce DIARRHEA and pyogenic infections. Pathogenic strains (virotypes) are classified by their specific pathogenic mechanisms such as toxins (ENTEROTOXIGENIC ESCHERICHIA COLI), etc. Alkalescens-Dispar Group,Bacillus coli,Bacterium coli,Bacterium coli commune,Diffusely Adherent Escherichia coli,E coli,EAggEC,Enteroaggregative Escherichia coli,Enterococcus coli,Diffusely Adherent E. coli,Enteroaggregative E. coli,Enteroinvasive E. coli,Enteroinvasive Escherichia coli
D005243 Feces Excrement from the INTESTINES, containing unabsorbed solids, waste products, secretions, and BACTERIA of the DIGESTIVE SYSTEM.
D005759 Gastroenteritis INFLAMMATION of any segment of the GASTROINTESTINAL TRACT from ESOPHAGUS to RECTUM. Causes of gastroenteritis are many including genetic, infection, HYPERSENSITIVITY, drug effects, and CANCER. Gastroenteritides
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D001257 Mamastrovirus A genus of small, circular RNA viruses in the family ASTROVIRIDAE. They cause GASTROENTERITIS and are found in the stools of several vertebrates including humans. Transmission is by the fecal-oral route and there are at least eight human serotypes. The type species is Human astrovirus. Human astrovirus,Human astroviruses,Mamastroviruses,astroviruses, Human
D060885 Multiplex Polymerase Chain Reaction Methods for using more than one primer set in a polymerase chain reaction to amplify more than one segment of the target DNA sequence in a single reaction. Multiplex Ligation-Dependent Probe Amplification,Multiplex PCR,Triplex PCR,Triplex Polymerase Chain Reaction,Multiplex Ligation Dependent Probe Amplification,PCR, Multiplex,PCR, Triplex

Related Publications

R Tolentino-Ruiz, and D Montoya-Varela, and M García-Espitia, and M Salas-Benito, and A Gutiérrez-Escolano, and C Gómez-García, and P Figueroa-Arredondo, and J Salas-Benito, and M De Nova-Ocampo
July 2013, Folia microbiologica,
R Tolentino-Ruiz, and D Montoya-Varela, and M García-Espitia, and M Salas-Benito, and A Gutiérrez-Escolano, and C Gómez-García, and P Figueroa-Arredondo, and J Salas-Benito, and M De Nova-Ocampo
May 2024, Food additives & contaminants. Part B, Surveillance,
R Tolentino-Ruiz, and D Montoya-Varela, and M García-Espitia, and M Salas-Benito, and A Gutiérrez-Escolano, and C Gómez-García, and P Figueroa-Arredondo, and J Salas-Benito, and M De Nova-Ocampo
February 2016, The plant pathology journal,
R Tolentino-Ruiz, and D Montoya-Varela, and M García-Espitia, and M Salas-Benito, and A Gutiérrez-Escolano, and C Gómez-García, and P Figueroa-Arredondo, and J Salas-Benito, and M De Nova-Ocampo
January 2014, PloS one,
R Tolentino-Ruiz, and D Montoya-Varela, and M García-Espitia, and M Salas-Benito, and A Gutiérrez-Escolano, and C Gómez-García, and P Figueroa-Arredondo, and J Salas-Benito, and M De Nova-Ocampo
February 2018, Veterinary parasitology,
R Tolentino-Ruiz, and D Montoya-Varela, and M García-Espitia, and M Salas-Benito, and A Gutiérrez-Escolano, and C Gómez-García, and P Figueroa-Arredondo, and J Salas-Benito, and M De Nova-Ocampo
August 2009, Journal of clinical microbiology,
R Tolentino-Ruiz, and D Montoya-Varela, and M García-Espitia, and M Salas-Benito, and A Gutiérrez-Escolano, and C Gómez-García, and P Figueroa-Arredondo, and J Salas-Benito, and M De Nova-Ocampo
October 2005, The Journal of molecular diagnostics : JMD,
R Tolentino-Ruiz, and D Montoya-Varela, and M García-Espitia, and M Salas-Benito, and A Gutiérrez-Escolano, and C Gómez-García, and P Figueroa-Arredondo, and J Salas-Benito, and M De Nova-Ocampo
October 2008, Veterinary microbiology,
R Tolentino-Ruiz, and D Montoya-Varela, and M García-Espitia, and M Salas-Benito, and A Gutiérrez-Escolano, and C Gómez-García, and P Figueroa-Arredondo, and J Salas-Benito, and M De Nova-Ocampo
May 2016, Journal of medical microbiology,
R Tolentino-Ruiz, and D Montoya-Varela, and M García-Espitia, and M Salas-Benito, and A Gutiérrez-Escolano, and C Gómez-García, and P Figueroa-Arredondo, and J Salas-Benito, and M De Nova-Ocampo
April 2023, Indian journal of gastroenterology : official journal of the Indian Society of Gastroenterology,
Copied contents to your clipboard!