Abstract
Gastrointestinal diseases and diarrhea are the major cause of morbidity and mortality in children in the developing world, including Mexico. In order to diagnose acute diarrhea and dysentery, four-multiplex polymerase chain reaction (mPCR) sets were designed and standardized in this study. Main virulence genes of relevant enteropathogens, that is,Escherichia coli pathotypes, Shigella spp., Salmonella enterica, Yersinia enterocolitica,Campylobacter jejuni and Aeromonas spp., were detected. Sixteen primer pairs were designed using extensive in silico analysis. As predicted in vitro, specific amplicons were obtained and four multiplex PCR sets were standardized. Our sixteen primer pairs and four multiplex PCR results demonstrate the simultaneous amplification of different pathogens according to the type of diarrhea in the same reaction. In the future, these methods might be used as rapid, sensitive and specific epidemiological and diagnostic tools. Key words: Gastrointestinal diseases, diarrhea, multiplex polymerase chain reaction, design, standardization, bioinformatics.
Highlights
Diarrhea occurs worldwide and is responsible for 4% of all deaths
The most common bacterial strains associated with diarrhea include Escherichia coli, Salmonella spp., Shigella spp., Yersinia enterocolitica, Vibrio cholerae and Campylobacter spp. (Gómez et al, 2009)
The aim of this study was to design in silico four-multiplex polymerase chain reaction (mPCR) sets and standardized to detect the main enteropathogens to establish the method as a new identification and epidemiological tool in our country
Summary
Diarrhea occurs worldwide and is responsible for 4% of all deaths. It is most commonly caused by gastrointestinal infections that kill around 2.2 million people globally each year, mostly children in developing countries according to the World Health Organization (WHO, 2013).In Mexico, gastrointestinal diseases are the main cause of medical consultation and death, so they are considered a public health problem. Diarrhea occurs worldwide and is responsible for 4% of all deaths. It is most commonly caused by gastrointestinal infections that kill around 2.2 million people globally each year, mostly children in developing countries according to the World Health Organization (WHO, 2013). In Mexico, gastrointestinal diseases are the main cause of medical consultation and death, so they are considered a public health problem. The most common bacterial strains associated with diarrhea include Escherichia coli, Salmonella spp., Shigella spp., Yersinia enterocolitica, Vibrio cholerae and Campylobacter spp. In Mexican clinical laboratories, diagnosis, identification and research focus mainly on classical pathogens, such as Salmonella, Shigella and Escherichia, so it is important to search for other underestimated relevant pathogens, such as Campylobacter, Yersinia and.
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