Simultaneous detection of multiple fly-borne bacterial pathogenic microorganisms by the reverse line blot hybridization assay

Abstract

Background: As a widespread health pest, flies can carry more than 100 kinds of path ogenic microbes to threat human health, resulting in a wide range of disease infection and transmission. The aim of this study was to develop a sensitive, reliable and rapid method for the simultaneous detection of multiple fly-borne bacterial pathogenic microorganisms, in order to effectively prevent and control fly-borne bacterial diseases. Results: PCR-RLB method could directly and accurately detect fly-borne bacteria species corresponding of 7 species-specific probes. The sensitivity of PCR-RLB was significantly higher (about 100 times) than that of PCR. At the same time, the membrane binding oligonucleotide species-specific probes prepared in RLB detection technology can be reused for detection of bacteria after washing with 0.5 M EDTA, which greatly improves the detection efficiency. This method was used to detect the intestinal pathogens carried by flies in four different areas of Lanzhou city. In 106 groups of samples from different areas, The numbers of carrying seven bacterial strains were 2 (S. aureus), 52 (S. flexneri), 0 (A. caviae), 3% (V. vulnificus), 56 (S. enterica), 1 (P. vulgaris) and 33 (Y. enterocolitica), respectively. Their proportions of 7 bacterial strains carried by houseflies were 1.23% ( S. aureus ), 32.1% ( S. flexneri ), 0% ( A. caviae ), 1.85% ( V. vulnificus ), 34.57% ( S. enterica ), 0.62% ( P. vulgaris ) and 20.37% ( Y. enterocolitica ), respectively. The results showed that there was a high correlation between the bacteria carrying rate of flies and the health environment in urban areas. S. enterica , S. flexneri and Y. enterocolitica accounted for the overwhelming majority of the seven pathogenic strains carried by houseflies. This indicated that houseflies played an important role in the transmission of intestinal infectious diseases. S. aureus and V. vulnificus were carried by houseflies near the hospital area indicates that hospitals should do well in killing and controlling flies and further strengthen the prevention and control of fly-borne bacterial diseases. Conclusion: The developed PCR-RLB assay have potential clinical application in the simultaneous detection of fly-borne bacterial species.