High Throughput Analysis of Bacterial Antibiotic Resistance Genes

Many authors have employed the WAVE System in facilitating Resistance Genotype Determination in context of infectious diseases. They found:

"Our results show that DHPLC is an efficient method for detecting mutations in genes that confer antibiotic resistance"¹

"Development and application of DHPLC will simplify considerably the identification of mutations associated with quinolone resistance. The data presented here provide evidence that DHPLC detects all of the functional mutations in QRDR encoding regions of the four genes grlA, gyrA, grlB and gyrB."²

"An advantage of the DHPLC system is that multiple methods may be programmed into the DHPLC system, thus enabling researchers to analyze different products in a walk-away fashion using the instrument’s autosampler and programmable column heater. Samples may also be recovered by using an optional fraction collector."³

"DHPLC analysis proved advantageous for the detection of novel and multiple mutations. DHPLC also provides a rapid, high-throughput alternative to LightCycler and SSCP for screening frequently occurring mutations."⁴

The following species and their corresponding candidate genes have been investigated using DHPLC on the WAVE System:

• S. aureus, S. pneumoniae, E. faecalis, E. faecium, E. coli, S. enterica, P. aeruginosa, B. fragilis and C. jejuni
• gyrA, gyrB, grlA and grlB genes
• B. anthracis, Y. pestis
• gyrA gene
• M. tuberculosis
• rpoB, katG, pncA, rpsL and embB genes

To find further details from all published papers and their abstracts in this field, go to the Journal Articles section of our Library and select the gene of interest.

References

1. Hurtle W, Lindler L, Fan W, Shoemaker D, Henchal E, Norwood D. (2003). Detection and identification of ciprofloxacin-resistant Yersinia pestis by denaturing high-performance liquid chromatography. J. Clin. Microbiol. 41, 3273-3283. Abstract Library Entry
2. Hannachi-M'Zali F, Ambler JE, Taylor CF, Hawkey PM (2002). Examination of single and multiple mutations involved in resistance to quinolones in Staphylococcus aureus by a combination of PCR and denaturing high-performance liquid chromatography (DHPLC) J. Antimicrob. Chemother. 50, 649-655. Abstract Library Entry
3. Cooksey RC, Morlock GP, Holloway BP, Limor J, Hepburn M. (2002). Temperature-mediated heteroduplex analysis performed by using denaturing high-performance liquid chromatography to identify sequence polymorphisms in Mycobacterium tuberculosis complex organisms. J. Clin. Microbiol. 40, 1610-1616. Abstract Library Entry
4. Eaves DJ, Liebana E, Woodward MJ, Piddock LJ. (2002). Detection of gyrA Mutations in Quinolone-Resistant Salmonella enterica by Denaturing High-Performance Liquid Chromatography. J. Clin. Microbiol. 40, 4121-4125. Abstract Library Entry