Identification of Pathogenic
            <i>Vibrio</i>
            Species by Multilocus PCR-Electrospray Ionization Mass Spectrometry and Its Application to Aquatic Environments of the Former Soviet Republic of Georgia

Whitehouse, C. A.; Baldwin, Carson; Sampath, Rangarajan; Blyn, Lawrence B.; Melton, Rachael; Feng, Li; Hall, Thomas A.; Harpin, Vanessa; Matthews, Heather; Тедиашвили, Марина; Jaiani, Ekaterine; Kokashvili, Tamar; Janelidze, Nino; Grim, Christopher J.; Colwell, Rita R.; Huq, Anwar

doi:10.1128/aem.01919-09

Cited by 18 publications

(19 citation statements)

References 10 publications

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“…The Ibis PCR-mass spectrometric technology has been used in both medical and environmental circumstances to detect and characterize both bacterial 26 and viral 27 organisms. It offers multiple advantages over other PCR-based assays in the evaluation of periprosthetic joint infection, including its broad survey capability, concomitant testing of common antibiotic resistance markers, relative speed, a small amount of source material necessary to perform the assay, and a user-friendly data readout.…”

Section: Resultsmentioning

confidence: 99%

Demonstration of Bacillus cereus in Orthopaedic-Implant-Related Infection with Use of a Multi-Primer Polymerase Chain Reaction-Mass Spectrometric Assay

Gallo¹,

Melton-Kreft²,

Nistico³

et al. 2011

Journal of Bone and Joint Surgery

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Section: Resultsmentioning

confidence: 99%

Demonstration of Bacillus cereus in Orthopaedic-Implant-Related Infection with Use of a Multi-Primer Polymerase Chain Reaction-Mass Spectrometric Assay

Gallo¹,

Melton-Kreft²,

Nistico³

et al. 2011

Journal of Bone and Joint Surgery

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“…The Ibis PCR‐MS technology has been used to detect and characterize both bacterial (Whitehouse et al , 2010) and viral organisms (Grant‐Klein et al , 2010), from both medical and environmental sources. It has multiple characteristics suggesting an excellent applicability to the diagnostic challenge frequently posed by prosthetic joint infection; in this case, it provided the first evidence of a multispecies infection, an observation subsequently confirmed by expanded culture, species‐specific PCR, RT‐PCR, and confocal microscopy using viability and FISH staining for targeted pathogens.…”

Section: Discussionmentioning

confidence: 99%

Characterization of a mixed MRSA/MRSE biofilm in an explanted total ankle arthroplasty

Stoodley

Conti

DeMeo

et al. 2011

FEMS Immunol Med Microbiol

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Bacterial biofilms have been observed in many prosthesis-related infections, and this mode of growth renders the infection both difficult to treat and especially difficult to detect and diagnose using standard culture methods. We (1) tested a novel coupled PCR-mass spectrometric (PCR-MS) assay (the Ibis T5000) on an ankle arthroplasty that was culture negative on preoperative aspiration and then (2) confirmed that the Ibis assay had in fact detected a viable multispecies biofilm by further micrographic and molecular examinations, including confocal microscopy using Live/Dead stain, bacterial FISH, and reverse-transcriptase-PCR (RT-PCR) assay for bacterial mRNA. The Ibis technology detected Staphylococcus aureus, Staphylococcus epidermidis, and the methicillin resistance gene mecA in soft tissues associated with the explanted hardware. Viable S. aureus were confirmed using RT-PCR, and viable cocci in the biofilm configuration were detected microscopically on both tissue and hardware. Species-specific bacterial FISH confirmed a polymicrobial biofilm containing S. aureus. A novel culture method recovered S. aureus and S. epidermidis (both methicillin resistant) from the tibial metal component. These observations suggest that molecular methods, particularly the new Ibis methodology, may be a useful adjunct to routine cultures in the detection of biofilm bacteria in prosthetic joint infection.

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“…PCR primers were designed to conserved regions within the selected target genes such that the targeted threat agent was clearly identified and differentiated from its near-neighbor species ( Table 1). Many of the primer pairs used in this assay have previously been used in other assays on the biosensor system described here [2,3,[11][12][13][14][15][16]. A complete analysis of each biocluster and the resolution provided by the assay is described below.…”

Section: A Comprehensive Biothreat Assaymentioning

confidence: 99%

Comprehensive Biothreat Cluster Identification by PCR/Electrospray-Ionization Mass Spectrometry

et al. 2012

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Technology for comprehensive identification of biothreats in environmental and clinical specimens is needed to protect citizens in the case of a biological attack. This is a challenge because there are dozens of bacterial and viral species that might be used in a biological attack and many have closely related near-neighbor organisms that are harmless. The biothreat agent, along with its near neighbors, can be thought of as a biothreat cluster or a biocluster for short. The ability to comprehensively detect the important biothreat clusters with resolution sufficient to distinguish the near neighbors with an extremely low false positive rate is required. A technological solution to this problem can be achieved by coupling biothreat group-specific PCR with electrospray ionization mass spectrometry (PCR/ESI-MS). The biothreat assay described here detects ten bacterial and four viral biothreat clusters on the NIAID priority pathogen and HHS/USDA select agent lists. Detection of each of the biothreat clusters was validated by analysis of a broad collection of biothreat organisms and near neighbors prepared by spiking biothreat nucleic acids into nucleic acids extracted from filtered environmental air. Analytical experiments were carried out to determine breadth of coverage, limits of detection, linearity, sensitivity, and specificity. Further, the assay breadth was demonstrated by testing a diverse collection of organisms from each biothreat cluster. The biothreat assay as configured was able to detect all the target organism clusters and did not misidentify any of the near-neighbor organisms as threats. Coupling biothreat cluster-specific PCR to electrospray ionization mass spectrometry simultaneously provides the breadth of coverage, discrimination of near neighbors, and an extremely low false positive rate due to the requirement that an amplicon with a precise base composition of a biothreat agent be detected by mass spectrometry.

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Identification of Pathogenic Vibrio Species by Multilocus PCR-Electrospray Ionization Mass Spectrometry and Its Application to Aquatic Environments of the Former Soviet Republic of Georgia

Cited by 18 publications

References 10 publications

Demonstration of Bacillus cereus in Orthopaedic-Implant-Related Infection with Use of a Multi-Primer Polymerase Chain Reaction-Mass Spectrometric Assay

Demonstration of Bacillus cereus in Orthopaedic-Implant-Related Infection with Use of a Multi-Primer Polymerase Chain Reaction-Mass Spectrometric Assay

Characterization of a mixed MRSA/MRSE biofilm in an explanted total ankle arthroplasty

Comprehensive Biothreat Cluster Identification by PCR/Electrospray-Ionization Mass Spectrometry

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