Ribosomal DNA-Directed PCR for Identification of Achromobacter ( Alcaligenes ) xylosoxidans Recovered from Sputum Samples from Cystic Fibrosis Patients

Abstract

The opportunistic human pathogen Achromobacter (Alcaligenes) xylosoxidans has been recovered with increasing frequency from respiratory tract culture of persons with cystic fibrosis (CF). However, confusion of this species with other closely related respiratory pathogens has limited studies to better elucidate its epidemiology, natural history, and pathogenic role in CF. Misidentification of A. xylosoxidans as Burkholderia cepacia complex is especially problematic and presents a challenge to effective infection control in CF. To address the problem of accurate identification of A. xylosoxidans, we developed a PCR assay based on a 16S ribosomal DNA sequence. In an analysis of 149 isolates that included 47 A. xylosoxidans and several related glucose-nonfermenting species recovered from CF sputum, the sensitivity and specificity of this PCR assay were determined to be 100 and 97%, respectively. The availability of this assay will enhance identification of A. xylosoxidans, thereby facilitating study of the pathogenic role of this species and improving infection control efforts in CF.

A Multilocus Sequence Typing Scheme Implies Population Structure and Reveals Several Putative Novel Achromobacter Species

“…However, our ongoing analyses of bacterial isolates recovered primarily from human specimens have identified a variety of "Achromobacter-like" strains that cannot be placed into one of these seven species with confidence, suggesting the existence of several additional distinct taxa. These strains, along with strains from each of the seven named species, tested positive in a previously described PCR assay targeting 16S rRNA gene sequences intended to be specific for A. xylosoxidans (11). Genotyping analyses of such strains using a rep-PCR typing method (3) indicated a high level of genetic diversity and identified several clusters, again suggestive of distinct species.…”

“…Achromobacter species are frequently misidentified by commercial identification systems, being most commonly mistaken for Alcaligenes, Bordetella, Burkholderia, Cupriavidus, Pandoraea, Ralstonia and Stenotrophomonas species (2,11). By using repetitive-element PCR (rep-PCR)-based genotyping employing the BOX A1R primer (BOX-PCR), we have noted remarkable diversity among strains we previously identified as A. xylosoxidans using a species-specific PCR assay designed to target signature 16S rRNA sequences in this species (11).…”

Infection Prevention and Control Guideline for Cystic Fibrosis: 2013 Update

“…have been attributed to contaminated disinfectant solutions, dialysis fluids, saline solution, and deionized water. 367 In addition, products used in nonhealthcare settings can become contaminated during manufacturing and can cause extensive outbreaks. These have included alcohol-tattoo ink contaminated with M. chelonae 368 and footbaths in nail salons contaminated with NTM spp.…”

Synergistic Effect of Combined Antibiotic and Methanol Extract of Eucalyptus camaldulensis leaf Against Staphylococcus aureus and Pseudomonas aeruginosa

“…Identification of selected MDR isolates were confirmed molecularly by investigation of 16S rRNA sequencing according to Liu et al, (2002) and Zhang et al, (2000).…”