1. A method for characterizing the antibiotic resistance of a microorganism, said method comprising the steps of:
a) providing a reference mass spectrum of an antimicrobial compound, its enzymatic modification product, or of a substrate compound of its modifying enzyme;
b) exposing a microorganism, a cell lysate thereof, or a growth medium supernatant thereof, to said antimicrobial compound or said substrate compound in aqueous liquid to thereby provide an exposed sample;
c) acquiring a mass spectrum of the exposed sample;
d) comparing the mass spectrum acquired in step c) with the reference mass spectrum of step a), and
e) determining from said comparison whether modification of said antimicrobial compound, its modification product or of said substrate compound has occurred following said exposure, and establishing that said microorganism is potentially resistant to said antimicrobial compound when said modification is observed.
2. Method according to claim 1, wherein said modification comprises enzymatic inactivation or enzymatic degradation of said antimicrobial compound or said substrate.
3. Method according to claim 2, wherein said enzymatic degradation is due to degradation by a beta-lactamase.
4. Method according to claim 3, wherein said beta-lactamase enzyme is selected from group A and D beta-lactamase enzymes according to the Ambler classification and beta-lactamase enzymes belonging to group 2 according to the Bush classification.
5. Method according to claim 4, wherein said beta lactamase enzyme is an extended-spectrum beta lactamase (ESBL).
6. Method according to any one of the preceding claims, wherein said microorganism is a suspected ESBL-producing microorganism, selected from Klebsiella pneumoniae, Escherichia coli, Klebsiella oxytoca and Proteus mirabilis.
7. Method according to claim 1, wherein said antimicrobial compound is a beta-lactam antibiotic.
8. Method according to claim 1, wherein said method is part of a method for characterization of an antibiotic modifying enzyme of a microorganism.
9. Method according to claim 8, wherein said method for characterization of said enzyme comprises the determination of the rate of modification of said antimicrobial compound or said substrate compound andor the rate of production of the enzymatic modification product of said antimicrobial compound or substrate compound to thereby determine the Michaelis-Menten (Km) constant and maximum reaction rate (Vmax) for said enzyme.
10. Method according to claim 1, wherein after step (b) said exposed sample is applied together with a matrix material to a mass spectrometric sample support and wherein said sample is dried on the sample support to produce a mass spectrometric sample for matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS).
11. Method according to claim 10, wherein said mass spectra are acquired using MALDI triple-quadrupole MS.
12. Method according to claim 1, wherein said exposed sample is an antimicrobial compound-exposed crude cell lysate of said microorganism.
13. Method according to claim 1, wherein in step (b) said microorganism is quantified by quantifying in said samples one or more structural biomolecules or metabolites derived from said microorganism.
14. Kit of parts for characterizing the antibiotic resistance of a microorganism comprising:
a) a lysis buffer for lysing a microorganism;
b) at least one antimicrobial compound or a substrate compound of an antimicrobial compound-modifying enzyme, and
c) a MALDI matrix material.
15. A system for characterizing the antibiotic resistance of a microorganism comprising:
at least one antimicrobial compound or a substrate of an antimicrobial compound-modifying enzyme;
a container for exposing a microorganism, a cell lysate thereof, or a growth medium supernatant thereof, to said at least one antimicrobial compound in aqueous liquid;
a lysis buffer for lysing said microorganism;
a MALDI matrix material;
a mass spectrometry device;
a reference mass spectrum of an antimicrobial compound, its enzymatic modification product, or of a substrate compound of a its modifying enzyme.
16. Method according to claim 1, wherein said mass spectrum is acquired using one of MALDI triple-quadrupole mass spectrometry, MALDI-TOF mass spectrometry and MALDI-FT-ICR mass spectrometry.
17. Method according to claim 1, wherein said exposed sample is a body fluid or body tissue sample of a human or animal subject suspected of carrying a microorganism a beta-lactam antibiotic resistance of which is to be characterized.
18. Method according to claim 7, the beta-lactam antibiotic is selected from the group consisting of penicillins, cephalosporins, cephamycins, carbapenems, ceftazidime, cefotaxime, ceftriaxone, cefpodoxime, and aztreonam.
19. Method according to claim 7, wherein the antibiotic compound further comprises a beta-lactamase inhibitor.
20. Method according to claim 19, wherein the beta-lactam antibiotic is one of amoxicillin and piperacillin and the beta-lactamase inhibitor is clavulanic acid or tazobactam, respectively.
21. Method according to claim 1, wherein a decrease in the level of antibiotic compound or the substrate compound or an increase in the level of a reaction product is measured by mass spectrometry and used as a measure for the modification.
22. Method according to claim 1, wherein the level of the antibiotic compound or substrate compound and the level of a reaction product are measured by mass spectrometry and the product-to-substrate ratio is used as a measure for the modification.
The claims below are in addition to those above.
All refrences to claim(s) which appear below refer to the numbering after this setence.
1. An embroidery data generator configured to generate embroidery data comprising:
a pattern storing unit configured to store plural types of pattern data configured for sewing one or more sub-patterns according to a predetermined stitch pattern, the one or more sub-patterns constituting an embroidery pattern; and
a control device configured to:
randomly extract pattern data configured for sewing the one or more sub-patterns from the plural types of pattern data stored in the pattern storing unit, and
assign extracted pattern data to the one or more sub-patterns.
2. The embroidery data generator according to claim 1, wherein the plural types of pattern data include plural types of surface region pattern data configured for sewing a surface region defined as an embroidery region according to a predetermined stitch pattern, and wherein the control device is configured to randomly extract pattern data, configured for sewing an inner side of an outline of the one or more sub-patterns as the surface region, from the plural types of surface region pattern data.
3. The embroidery data generator according to claim 2, wherein when the embroidery pattern is formed of two or more sub-patterns, the control device is further configured to assign different surface region pattern data to adjacent sub-patterns.
4. The embroidery data generator according to claim 1, wherein the plural types of pattern data include plural types of outline pattern data configured for sewing an outline of the one or more sub-patterns according to a predetermined stitch pattern, and wherein the control device is further configured to randomly extract pattern data for sewing the outline of the one or more sub-patterns from the plural types of outline pattern data.
5. The embroidery data generator according to claim 1, further comprising a category specifying unit configured to specify one of plural categories, wherein the plural types of pattern data stored in the pattern storing unit is classified into plural categories, and wherein the control device is further configured to randomly extract pattern data for sewing the one or more sub-patterns from pattern data belonging to a category selected by the category specifying unit.
6. The embroidery data generator according to claim 1, further comprising an edit unit configured to edit parameters for modifying size andor shape of the predetermined stitch pattern.
7. The embroidery data generator according to claim 1, further comprising a color storing unit configured to store multiple entries of predefined color information, wherein the control device is further configured to:
randomly extract a color of thread, used for sewing the one or more sub-patterns based on the plural types of pattern data, from color information stored in the color storing unit, and
assign randomly extracted color to each of the one or more sub-patterns.
8. The embroidery data generator according to claim 7, wherein the plural types of pattern data include plural types of surface region pattern data configured for sewing a surface region, defined as an embroidery region, according to a predetermined stitch pattern, and wherein the control device is further configured to:
randomly extract a color of thread, used for sewing a surface region of the one or more sub-patterns based on the plural types of surface region pattern data, from color information stored in the color storing unit, and
assign different colors to adjacent sub-patterns.
9. The embroidery data generator according to claim 1, further comprising a display unit configured to display information pertaining to a sewing operation, wherein the control device is further configured to display the embroidery pattern with the stitch pattern defined in extracted pattern data.
10. A non-transitory computer readable storing medium storing computer readable instructions that, when executed by a control device of an embroidery data generator provided with a pattern storing unit configured to store plural types of pattern data configured for sewing an embroidery pattern formed of one or more sub-patterns according to a predetermined stitch pattern, cause the control device to:
randomly extract pattern data configured for sewing the one or more sub-patterns from the plural types pattern data stored in the pattern storing unit, and
assign extracted pattern data to the one or more sub-patterns.
11. The medium according to claim 10, wherein the plural types of pattern data include plural types of surface region pattern data configured for sewing a surface region defined as an embroidery region according to a predetermined stitch pattern, wherein the instructions further cause the control device to randomly extract pattern data configured for sewing an inner side of an outline of the one or more sub-patterns as the surface region from the plural types of surface region pattern data.
12. The medium according to claim 11, wherein the instructions further cause the control device to assign different surface region pattern data to adjacent sub-patterns.
13. The medium according to claim 10, wherein the plural types of pattern data include plural types of outline pattern data configured for sewing an outline of the one or more sub-patterns according to a predetermined stitch pattern, and wherein the instructions further cause the control device to randomly extract pattern data configured for sewing the outline of the one or more sub-patterns from the plural types of outline pattern data.
14. The medium according to claim 10, wherein the embroidery data generator further comprises a category specifying unit configured to specify one of plural categories, wherein the plural types of pattern data stored in the pattern storing unit is classified into plural categories, and wherein the instructions further cause the control device to randomly extract pattern data for sewing the one or more sub-patterns from pattern data belonging to a category selected by the category specifying unit.
15. The medium according to claim 10, wherein the instructions further cause the control device to edit parameters for modifying a size andor a shape of the predetermined stitch pattern upon receiving an input operation.
16. The medium according to claim 10, wherein the embroidery data generator further comprises a color storing unit configured to store multiple entries of predefined color information, wherein the instructions further cause the control device to:
randomly extract a color of thread, used for sewing the one or more sub-patterns based on the plural types of pattern data, from color information stored in the color storing unit, and
assign randomly extracted color to each of the one or more sub-patterns.
17. The medium according to claim 16, wherein the plural types of pattern data include plural types of surface region pattern data configured for sewing a surface region defined as an embroidery region according to a predetermined stitch pattern, and wherein the instructions further cause the control device to:
randomly extract a color of thread, used for sewing a surface region of the one or more sub-patterns based on the plural types of surface region pattern data, from color information stored in the color storing unit, and
assign different colors to adjacent sub-patterns.
18. The medium according to claim 10, wherein the embroidery data generator further comprises a display unit configured to display information pertaining to a sewing operation, wherein the instructions further cause the control device to display the embroidery pattern in the stitch pattern defined in extracted pattern data.
19. A sewing machine comprising:
a sewing unit configured to be capable of sewing a workpiece based on embroidery data;
a pattern storing unit configured to store plural types of pattern data configured for sewing one or more sub-patterns according to a predetermined stitch pattern, the one or more sub-patterns constituting an embroidery pattern; and
a control device configured to:
randomly extract pattern data, configured for sewing the one or more sub-patterns, from the plural types of pattern data stored in the pattern storing unit,
assign extracted pattern data to the one or more sub-patterns, and
control the sewing unit to sew an embroidery pattern on the workpiece, the embroidery pattern formed of the one or more sub-patterns having been assigned the pattern data.