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Background Bone and Joint Infections (BJI) have high morbidity and are difficult to treat infections. Culture-based diagnosis is limited in its ability to recover fastidious bacteria and because several organisms can be involved; culture times of up to two weeks may be necessary for certain bacteria. The sensitivity of culture is also negatively impacted by antibiotics received before surgery. Alternatively, molecular methods offer a promising improvement for the diagnosis of BJI. The goal of this study was to evaluate a development version of Biofire® Bone and Joint Infection (BJI) Panel (bioMerieux SA, BioFire Diagnostics, LLC) using synovial fluid samples. Methods 121 synovial fluid specimens were collected from patients with suspected bone and joint infection in a pilot evaluation. All specimens were collected and tested in culture by the sites using their standard of care practices; in parallel, a leftover volume of 200 µL was tested on the BJI panel. BJI panel results were then compared with culture and discordant results were investigated using a comparator assay (PCR/sequencing). Results 49 synovial fluid specimens (40%) were positive by culture vs. 72 with the BJI panel (59%). Of the 97 positive detections by the BJI panel, 58 were concordant with culture; the 39 additional organism detections were in majority confirmed by PCR/sequencing. Lastly, two false negative results corresponding to the same sample are under investigation. Conclusion The BJI Panel was able to identify most of the pathogens detected by culture. The majority of additional detections observed were confirmed by PCR/sequencing. While sites are currently enrolling more synovial fluids samples, these preliminary data suggest that a multiplexed molecular test may be more sensitive than culture to detect pathogens in synovial fluid specimens. The data presented in this abstract have not been reviewed by FDA or other regulatory agencies for In Vitro Diagnostic use. Disclosures B. Pons, bioMerieux: Employee, Salary. C. Jay, bioMerieux: Employee, Salary. T. Martin, bioMerieux: Employee, Salary. I. Sothier, bioMerieux: Employee, Salary. H. Savelli, bioMerieux: Employee, Salary. B. Kensinger, bioFire a bioMerieux company: Employee, Salary. F. Laurent, BioFire (bioMerieux company): Investigator, Research support. L. Abad, BioFire (bioMerieux company): Investigator, Research support. C. Murphy, BioFire (bioMerieux company): Investigator, Research support. A. Craney, BioFire (bioMerieux company): Investigator, Research support. B. Schmitt, BioFire (bioMerieux company): Investigator, Research support. A. Waggoner, BioFire (bioMerieux company): Investigator, Research support. S. Butler-Wu, BioFire (bioMerieux): Investigator, Research support. C. Costales, BioFire (bioMerieux company): Investigator, Research support. J. Bien-Bard, BioFire (bioMerieux): Investigator, Research support. J. Mestas, BioFire (bioMerieux): Investigator, Research support. J. Esteban, BioFire (bioMerieux): Investigator, Research support. L. Salar-Vidal, BioFire (BioMerieux company)): Investigator, Research support. A. Harrington, BioFire (bioMerieux company): Investigator, Research support. S. Collier, BioFire (BioMerieux Company): Investigator, Research support. A. Leber, BioFire (bioMerieux company): Investigator, Research support. K. Everhart, BioFire (bioMerieux company): Investigator, Research support. J. M. Balada-Llasat, BioFire (bioMerieux company): Investigator, Research support. J. Horn, BioFire (bioMerieux company): Investigator, Research support. S. Magro, bioMerieux: Employee, Salary. K. Bourzac, BioFire a bioMerieux company: Employee, Salary.

A new and efficient strategy for labeling of nucleic acids prior to their hybridization on high density DNA chip has been developed. Our approach which combines the fragmentation and the labeling is based on the reactivity of the terminal phosphates of cleaved DNA and RNA fragments with a reporter molecule bearing aryldiazomethane group.

An efficient strategy for nucleic acid labeling and analysis on deoxyribonucleic acid (DNA) chips has been developed. This approach, which combines the fragmentation and the labeling steps, is based on the reactivity of the phosphates of DNA and ribonucleic acid (RNA) fragments and is using reporter molecules bearing a bromomethyl- or aryldiazomethane-reactive group. In this chapter, we describe the preparation of the reactive label and protocols for efficient labeling of any nucleic acid sequence, DNA or RNA, prior to their hybridization, detection, and analysis on DNA chips.

An efficient strategy for nucleic acid labeling and analysis on deoxyribonucleic acid (DNA) chips has been developed. This approach, which combines the fragmentation and the labeling steps, is based on the reactivity of the phosphates of DNA and ribonucleic acid (RNA) fragments and is using reporter molecules bearing a bromomethyl- or aryldiazomethane-reactive group. In this chapter, we describe the preparation of the reactive label and protocols for efficient labeling of any nucleic acid sequence, DNA or RNA, prior to their hybridization, detection, and analysis on DNA chips.

DNA and RNA labeling and detection are key steps in nucleic acid-based technologies, used in medical research and molecular diagnostics. We report here the synthesis, reactivity, and potential of a new type of labeling molecule, m-(N-Biotinoylamino)phenylmethyldiazomethane (m-BioPMDAM), that reacts selectively and efficiently with phosphates in nucleotide monomers, oligonucleotides, DNA, and RNA. This molecule contains a biotin as detectable unit and a diazomethyl function as reactive moiety. We demonstrate that this label fulfills the requirements of stability, solubility, reactivity, and selectivity for hybridization-based analysis and especially for detection on high-density DNA chips.

We show here a new and efficient aqueous chemistry for labeling of any class of nucleic acids for their detection on DNA chip. The labels contain a diazo function as reactive moiety and biotin as detectable unit. The highly selective reaction of diazo group on the phosphate does not disrupt base pairing recognition and hybridization specificity.