Bacterial response
A technology of bacteria and strains, applied in bioreactor/fermenter combination, special-purpose bioreactor/fermenter, laboratory container, etc.
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0090] Antibiotic sensitivity can be measured at the molecular level by detecting transcriptional differences between susceptible and resistant bacteria in response to antibiotic exposure. These transcriptional differences can be illustratively discovered using RNA-sequencing or cDNA microarray analysis. The large multiplex and reverse transcription capabilities of multiplex systems such as the FilmArray system, as described above, can facilitate antibiotic susceptibility measurements for multiple bacteria and antibiotics. In this example, specific and general bacteria-antibiotic combinations were targeted after exposure to antibiotics to determine whether differences could be detected between susceptible and resistant strains. It is understood that such methods can be extended to other antibiotics or mixtures of antibiotics.
[0091] In prior art methods, both sensitive and resistant cultures were grown to early logarithmic phase and then exposed to antibiotics or no exposur...
Embodiment 2
[0114] The benchtop multiplex experiments described above demonstrate the feasibility of using measurements of cellular RNA concentrations of general antibiotic resistance genes and / or specific antibiotic resistance genes as antibiotic resistance tests. The ability to measure the concentration of, or detect the presence of, bacterial RNA transcripts is hampered by the fact that large numbers of these transcripts are present at concentrations well below one transcript / cell (Bartholomaus et al.). A practical consequence of this is that the concentration (copy / cell) of cellular genomic DNA (at least 1 copy / cell) often far exceeds the concentration of any cellular RNA transcript (often [RNA]). Under these conditions, removal of genomic DNA can help facilitate detection of RNA signals.
[0115] DNA removal can be accomplished using a variety of strategies, illustratively by modifying cell lysis conditions to enable selective release of RNA, modifying nucleic acid purification to s...
Embodiment 3
[0127] Although the FilmArray system is mentioned herein, other systems are suitable for the methods used herein. Certain embodiments of the invention may also relate to or include a PCR system configured to call from an amplification curve or a melting curve or a combination thereof. An illustrative example is described in US Patent No. 8,895,295, which has been incorporated by reference, for use with pouch 510 or similar embodiments. It should be understood, however, that the embodiments described in US Patent No. 8,895,295 are illustrative only and that other systems may be used in light of the present disclosure. For example, refer to Figure 10 , shows a block diagram of an illustrative system 700 including a control element 702, a thermal cycling element 708, and an optical element 710 in accordance with an exemplary aspect of the present disclosure.
[0128] In at least one embodiment, a system can include at least one PCR reaction mixture contained in a sample vessel...
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More