Development, validation, and application of PCR primers for detection of tetracycline efflux genes of gram-negative bacteria.

Applied and Environmental Microbiology
Rustam I AminovRoderick I Mackie

Abstract

Phylogenetic analysis of tetracycline resistance genes, which confer resistance due to the efflux of tetracycline from the cell catalyzed by drug:H(+) antiport and share a common structure with 12 transmembrane segments (12-TMS), suggested the monophyletic origin of these genes. With a high degree of confidence, this tet subcluster unifies 11 genes encoding tet efflux pumps and includes tet(A), tet(B), tet(C), tet(D), tet(E), tet(G), tet(H), tet(J), tet(Y), tet(Z), and tet(30). Phylogeny-aided alignments were used to design a set of PCR primers for detection, retrieval, and sequence analysis of the corresponding gene fragments from a variety of bacterial and environmental sources. After rigorous validation with the characterized control tet templates, this primer set was used to determine the genotype of the corresponding tetracycline resistance genes in total DNA of swine feed and feces and in the lagoons and groundwater underlying two large swine production facilities known to be impacted by waste seepage. The compounded tet fingerprint of animal feed was found to be tetCDEHZ, while the corresponding fingerprint of total intestinal microbiota was tetBCGHYZ. Interestingly, the tet fingerprints in geographically distant waste l...Continue Reading

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Oct 15, 2011·Microbial Ecology·Sepideh PakpourMartin R Chénier
Jan 9, 2009·Applied Microbiology and Biotechnology·Xu-Xiang ZhangHerbert H P Fang
Nov 29, 2012·Applied Microbiology and Biotechnology·Miaomiao LiuMin Yang
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