| 摘要 |
The present invention provides a method for detecting and characterising a microorganism in aclinical sample, said method comprising: a) introducing a clinical sample to a first culture vessel containing culture medium; b (i) optionally preculturing said clinical sample in said first culture vessel; (ii) optionally removing a portion of the clinical sample/medium mixture or, if precultured, the clinical sample culture from said first culture vessel, and introducing said portion to a second culture vessel containing culture medium, and optionally preculturing said portion in said second culture vessel; c) removing a test aliquot from said first and/or second culture vessel, and culturing or continuing to culture said clinical sample and/or portion in said first and/or second culture vessel; d) separating DNA from said test aliquot; e) performing nucleic acid tests on said DNA to identify the microorganism and to detect the presence or absence of one or more genetic antimicrobial resistance markers in said microorganism, wherein said nucleic acid tests are performed using: i) one or more nucleic acid probes and/or primers for microbial identification, a said probe or primer being capable of hybridising specifically to, or a said primer being capable of selectively amplifying, a nucleotide sequence which is identificatory of a given microorganism; and ii) one or more nucleic acid probes and/or primers for antimicrobial resistance marker detection, a said probe or primer being capable of hybridising specifically to, or a said primer being capable of selectively amplifying, a nucleotide sequence representing a genetic antimicrobial resistance marker; and it is detected whether or not said probes and/or primers have hybridised to said DNA and/or said primers have been extended (e.g. an amplification reaction has taken place); and f) if a microorganism is identified in step (e), performing an antimicrobial susceptibility test on said cultured clinical sample and/or portion from step (c), wherein microbial growth in said antimicrobial susceptibility test is monitored by assessing growth or markers for growth, and wherein the type and concentration of antimicrobial agents used in said antimicrobial susceptibility test is determined by the identity of the microorganism and antimicrobial resistance markers detected in step (e), and optionally continuing to culture said clinical sample and/or portion in said first and/or second culture vessel or g) if no microorganism strain is identified in step (e), further culturing said clinical sample and/or portion to enable further microbial identification and antimicrobial susceptibility tests to be performed to identify the microorganism and determine its antimicrobial resistance profile.A device for performing the method of the invention is also provided. |
