ArticleApplied and environmental microbiology2006
Internally controlled real-time PCR method for quantitative species-specific detection and vapA genotyping of Rhodococcus equi.
Article in Applied and environmental microbiology, 2006. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 17 papers.
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Who cites it
17 citing papers in PubMed, 52 citations in OpenAlex.
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- Assessment of listing and categorisation of animal diseases within the framework of the Animal Health Law (Regulation (EU) No 2016/429): antimicrobial-resistantEFSA journal. European Food Safety Authority · 2022Article
- Antimicrobial Resistance Spectrum Conferred by pRErm46 of Emerging Macrolide (Multidrug)-Resistant Rhodococcus equi.Journal of clinical microbiology · 2021Article
- Molecular characterization ofVeterinary world · 2017Article
- Development and evaluation of the internal-controlled real-time PCR assay for Rhodococcus equi detection in various clinical specimens.The Journal of veterinary medical science · 2016Article
- VirS, an OmpR/PhoB subfamily response regulator, is required for activation of vapA gene expression in Rhodococcus equi.BMC microbiology · 2014Article
- Simultaneous detection of Streptococcus pneumoniae, S. mitis, and S. oralis by a novel multiplex PCR assay targeting the gyrB gene.Journal of clinical microbiology · 2013Article
- Density interactions among Streptococcus pneumoniae, Haemophilus influenzae and Staphylococcus aureus in the nasopharynx of young Peruvian children.The Pediatric infectious disease journal · 2013Article
- Rhodococcus equi venous catheter infection: a case report and review of the literature.Journal of medical case reports · 2011Article
- Identification of atypical Rhodococcus-like clinical isolates as Dietzia spp. by 16S rRNA gene sequencing.Journal of clinical microbiology · 2010Article
- Mycoplasma agalactiae p40 Gene, a novel marker for diagnosis of contagious agalactia in sheep by real-time PCR: assessment of analytical performance and in-house validation using naturally contaminated milk samples.Journal of clinical microbiology · 2009Article
- Development of quantitative real-time PCR assays for detection and quantification of surrogate biological warfare agents in building debris and leachate.Applied and environmental microbiology · 2007Article
- Quantitative detection of Clostridium tyrobutyricum in milk by real-time PCR.Applied and environmental microbiology · 2007Article
- Use of Serial Quantitative PCR of the vapA Gene of Rhodococcus equi in Feces for Early Detection of R. equi Pneumonia in Foals.Journal of veterinary internal medicineArticle
- Estimating the Sensitivity and Specificity of Real-Time Quantitative PCR of Fecal Samples for Diagnosis of Rhodococcus equi Pneumonia in Foals.Journal of veterinary internal medicineArticle
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Authors and funding
9 authors at 5 institutions in 4 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
We developed a novel quantitative real-time PCR (Q-PCR) method for the soil actinomycete Rhodococcus equi, an important horse pathogen and emerging human pathogen. Species-specific quantification was achieved by targeting the chromosomal monocopy gene choE, universally conserved in R. equi. The choE Q-PCR included an internal amplification control (IAC) for identification of false negatives. A second Q-PCR targeted the virulence plasmid gene vapA, carried by most horse isolates but infrequently found in isolates from other sources. The choE-IAC and vapA assays were 100% sensitive and specific as determined using 178 R. equi isolates, 77 nontarget bacteria, and a panel of 60 R. equi isolates with known vapA+ and vapA-negative (including vapB+) plasmid genotypes. The vapA+ frequency among isolate types was as follows: horse, 85%; human, 20%; bovine and pig, 0%; others, 27%. The choE-IAC Q-PCR could detect up to one genome equivalent using R. equi DNA or 100 bacteria/ml using DNA extracted from artificially contaminated horse bronchoalveolar lavage (BAL) fluid. Quantification was linear over a 6-log dynamic range down to approximately 10 target molecules (or 1,000 CFU/ml BAL fluid) with PCR efficiency E of >0.94. The vapA assay had similar performance but appeared unsuitable for accurate (vapA+) R. equi quantification due to variability in target gene or plasmid copy number (1 to 9). The dual-reaction Q-PCR system here reported offers a useful tool to both medical and veterinary diagnostic laboratories for the quantitative detection of R. equi and (optional) vapA+ "horse-pathogenic" genotype determination.
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