Identification of bacterial intestinal pathogens by a PCR-Reverse dot blot procedure

Ho Thi Thanh Thuy; Lao Duc Thuan; Truong Kim Phuong; Le Huyen Ai Thuy

6 (1) 2016

Identification of bacterial intestinal pathogens by a PCR-Reverse dot blot procedure

Author - Affiliation:

Ho Thi Thanh Thuy - Viet A Technology Corporation , Vietnam

Lao Duc Thuan - Ho Chi Minh City Open University , Vietnam

Truong Kim Phuong - Ho Chi Minh City Open University , Vietnam

Le Huyen Ai Thuy - Ho Chi Minh City Open University , Vietnam

Corresponding author: Le Huyen Ai Thuy - thuy.lha@ou.edu.vn

Abstract

Intestinal infections which are the important public health concern worldwide, are caused by the bacterial intestinal pathogens. The aim at our study is to develop a simultaneous, rapid, sensitive and specific diagnostic assay by using a combined PCR-Reverse dot blot method for the identification of pathogen strains, including Bacillus cereus, Clostridium botulinum, Clostridium perfringen, Staphylococcus aureus, Listeria monocytogenes, Escherichia coli O157:H7, Salmonella spp., Shigella spp., Vibrio cholerae, Vibrio parahaemolyticus, Yersinia enterocolitica and Brucella spp.
Based on the 16S and 23S DNA regions, the two sets of universal primers and twelve specific probes were obtained for amplification and specific detection of those twelve bacterial species. The initial experimental results using bacterial cultures and 50 clinical samples confirmed the in silico hypothesis that was previously established in universal primers and probes design as well as identified with some basic conditions for PCR and Reverse Dot Blot hybridization reactions. Thus, this procedure being further tested for the other kinds of samples such as fecal samples or foods.

Keywords

16S RDNA; 23S rDNA; PCR; Reverse dot blot; bacterial intestinal pathogens

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