Identification of Sphingomonads on the Basis of Polymerase Chain Reaction Amplified 16S rRNA Gene

Abstract

Abstract Sphingomonas is a genus that is basically of environmental origin but can also be associated with health hazards, especially in the hospital environment where there is a great need to properly monitor water sources. The abundance and frequent isolation of derivatives of yellow pigmented colonies from drinking water samples in Lebanon—where an intermittent mode of supply is employed, and which induces frequent biofilm sloughing—necessitated the establishment of a rapid and feasible assay to screen specifically for sphingomonads. In this study, 50 isolates recovered from drinking water with yellow- to orange-pigmented colonies were used to establish a polymerase chain reaction-based (PCR-based) screening assay. The use of sphingomonad specific modified primers gave one common band with a size of 320 bp in all resumptive and sequence confirmed sphingomonads. However, no amplification was observed with Escherichia coli, Staphylococcus aureus, and Pseudomonas aeruginosa. Applying the PCR-based assay described in this paper increased both the efficiency and the reliability of screening for sphingomonads in water samples, thereby minimizing related risk factors.