Master mix stocks were prepared containing final concentrations of 5 μM protein and 10X SYPRO Orange (Pierce Thermo) in NMR buffer and contained either 0 μM, 5 μM, 10 μM, or 50 μM doripenem or imipenem (Sigma-Aldrich) and were incubated overnight to ensure reactions were complete. Each master mix was dispensed in 50 μL triplicate aliquots into a 96-well skirted PCR plate (Bio-Rad) and subsequently sealed with a clear thermal-seal film to prevent evaporation. The sample plate was spun down at 2,000 RPM for 2 minutes in an Eppendorf 5430 R centrifuge equipped with a A-2-MTP rotor. DSF experiments were conducted on a CFX96 Touch Real-Time PCR system (Bio-Rad) using the HEX channel for both excitation and emission. The experimental program collected fluorescence intensity at 0.5 °C increments with a 5 second equilibration step between each temperature. The fluorescence intensity (RFU) data points were normalized by the highest value, and then fit to Boltzmann Sigmoidal curve using Prism GraphPad, where the inflection point indicated the melted temperature (Tm). Statistical uncertainties were estimated as the standard deviation of the inflection point for each triplicate data set.
Doripenem
Manufactured by Merck Group
Sourced in United States
Doripenem is a laboratory product developed by Merck Group for research purposes. It is a carbapenem antibiotic used as a standard for antimicrobial susceptibility testing. Doripenem is a broad-spectrum antibiotic with activity against various gram-positive and gram-negative bacteria.
Automatically generated - may contain errors
Lab products found in correlation
Imipenem, by Merck Group (7 mentions)
Meropenem, by Merck Group (5 mentions)
Amikacin, by Merck Group (4 mentions)
Ciprofloxacin, by Merck Group (4 mentions)
Colistin, by Merck Group (4 mentions)
Aztreonam, by Merck Group (3 mentions)
Ceftazidime, by Merck Group (3 mentions)
Levofloxacin, by Merck Group (3 mentions)
Cefotaxime, by Merck Group (2 mentions)
Biapenem, by Merck Group (2 mentions)
Colistin sulfate, by Merck Group (2 mentions)
Tigecycline, by Pfizer (2 mentions)
Cefepime, by Merck Group (2 mentions)
Gentamicin, by Merck Group (2 mentions)
Tigecycline, by Thermo Fisher Scientific (2 mentions)
Cfx96 touch real time pcr system, by Bio-Rad (1 mentions)
Ertapenem, by Merck Group (1 mentions)
Tyloxapol, by Merck Group (1 mentions)
Ethambutol, by Merck Group (1 mentions)
Rifampicin rif, by Merck Group (1 mentions)
13 protocols using doripenem
1
Thermal Stability of Antibiotics via DSF
2
Antimicrobial Susceptibility Profiling
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mtb H37Rv, Msm mc2-155 and Msm PM965 (ept-1 rpsL4 ΔblaS1), a mutant strain deficient for the major beta-lactamase BlaS (Raymond et al., 2005 (link)), were used in this study. Bacteria were grown in Middlebrook 7H9 medium (BD Biosciences) or in Middlebrook 7H10 medium (BD Biosciences), supplemented with 0.2% or 0.5% of glycerol, respectively. Both media were supplemented with 10% oleic acid-albumin-dextrose-catalase (BD Difco) for Mtb and 0.5% glucose for Msm. Tyloxapol (Sigma-Aldrich) was added to liquid medium to a final concentration of 0.05% to prevent clump formation. Stocks of amoxicillin (AMX), biapenem (BIA), cefotaxime (CTX), doripenem (DOR), ertapenem (ETP), ethambutol (EMB), meropenem (MEM) and vancomycin (VAN) (Sigma-Aldrich) were prepared in purified water. Potassium clavulanate (CLA) (Sigma-Aldrich) was prepared in phosphate buffer pH 6.0, 0.1 M. Isoniazid (INH) and rifampicin (RIF) (Sigma-Aldrich) were prepared in dimethyl sulfoxide or methanol, respectively.
3
Polymyxin B and Lipid Vesicle Interactions
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Polymyxin B (PMB; lot number 20120204), amikacin (lot number 058K0803), aztreonam (batch number MKCH8931), and doripenem (lot number 0137Y01) were purchased from Sigma-Aldrich and their solutions were prepared in sterilized Milli-Q water (Millipore, Australia). Phytantriol (98%, 3,7,11,15-tetramethylhexadecane-1,2,3-triol), poly(ethylene oxide)-poly(propylene oxide)-poly(ethylene oxide) triblock copolymer (Pluronic F127), lipid A (diphosphoryl from Escherichia coli F583), Octadecyl Rhodamine B Chloride (R18), Dulbecco’s modified Eagle’s medium (DMEM, 4.5 g/L glucose), fetal bovine serum (FBS) and [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (tetrazole)] (MTT) were purchased from Sigma-Aldrich; 1,2-dipalmitoyl-d62-sn-glycero-3-phosphocholine (d-DPPC) was purchased in powder form from Avanti Polar Lipids Inc. CellBriteTM Fix 488 membrane stain was purchased from Biotium. Milli-Q water 18.2 MΩ ∙ cm and D2O (99.99 atom%) were used in all experiments for preparation of buffers and solutions. All chemicals were used as received without further purification. The Human embryonic kidney 293T cells (ATCC HEK-293T) was the gift from Dr Thomas Naderer, Monash University.
4
Antibiotics Acquisition and Preparation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
All culture media were purchased from Becton Dickinson & Co. Difco (Franklin Lakes, NJ, USA). Colistin sulfate, minocycline hydrochloride, doripenem, and tobramycin were obtained from Sigma-Aldrich, (Saint Louis, MO, USA). Ciprofloxacin, and levofloxacin were purchased from Siam Bheasach Co., Ltd. (Bangkok, Thailand). Tigecycline was purchased from Pfizer Inc. (Philadelphia, PA, USA). Imipenem was obtained from Merck Sharp & Dohme Corp. (Elkton, VA, USA). Meropenem was obtained from M&H manufacturing Co., Ltd. (Samutprakarn, Thailand). Fosfomycin was obtained from Meiji Seikakaisna, Ltd. (Tokyo, Japan).
5
Antimicrobial Susceptibility Testing Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Standard amikacin, ciprofloxacin, doxycycline, ertapenem, gentamicin, imipenem (U.S. Pharmacopeia, Rockville, MD, USA), ampicillin, cephalothin, cefuroxime, ceftriaxone, ceftazidime, colistin sulfate, doripenem, meropenem, trimethoprim/sulfamethoxazole (Sigma-Aldrich, St. Louis, MO, USA), fosfomycin (Ercros, Barcelona, Spain), and tigecycline (Pfizer, New York, NY, USA) were used for antimicrobial susceptibility testing. The minimum inhibitory concentration (MIC) determinations and susceptibility interpretation criteria followed the Clinical Laboratory and Standard Institute (CLSI) and Federal Drug Administration (FDA) standards [11 ,12 (link)]. The MICs of the drugs, except tigecycline and colistin, were measured by agar dilution in Mueller–Hinton agar (Oxoid, Basingstoke, UK) according to CLSI recommendations.11 For fosfomycin susceptibility, glucose-6-phosphate (25 mg/mL) was added to the agar plate. tigecycline and colistin MICs were determined by microdilutions in freshly prepared cation-adjusted Mueller–Hinton broth (CAMHB). E. coli ATCC 25922 was used as the control strain [13 (link)]. The MICs of other agents were determined using the custom-designed panels for Gram-negative bacilli (Sensititre, Thermo Fisher Scientific, Oakwood Village, OH, USA).
6
Antimicrobial Susceptibility Testing Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Minimal inhibitory concentrations were determined according to Clinical and Laboratory Standards Institute® (CLSI) M07-A10 (CLSI, 2015 ) in 96-well plates. Breakpoints interpretation were made according to the M100 Performance Standards for Antimicrobial susceptibility testing 28th edition (CLSI, 2019 ). Antibiotics included were amikacin (Sigma Aldrich A1774), gentamicin (Sigma Aldrich G3632), aztreonam (Sigma Aldrich PZ0038), ceftazidime (Sigma Aldrich C3809), cefepime (Sigma Aldrich PHR1763), ciprofloxacin (Sigma Aldrich 17850), levofloxacin (Sigma Aldrich 28266), doripenem (Sigma Aldrich 32138), imipenem (Sigma Aldrich I0160), meropenem (Sigma Aldrich M2574), colistin (Sigma Aldrich C4461), and piperacillin/tazobactam (Sigma Aldrich P8396/T2820). P. aeruginosa ATCC® 27853 was used as control as according to CLSI (Supplementary Table S1 ).
7
Antimicrobial Susceptibility Testing of P. aeruginosa
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Minimal inhibitory concentrations (MICs) of amikacin (Sigma-Aldrich, St. Louis, MO), aztreonam (Sigma), cefepime (Sigma), ceftazidime (Sigma), ciprofloxacin (Sigma), doripenem (Sigma), gentamicin (Amresco Inc., Solon, OH), imipenem (Sigma), levofloxacin (Sigma), and meropenem (Sumitomo Pharmaceuticals Co., Ltd., Osaka, Japan) were determined in duplicate by the agar dilution method according to the recommendations of the CLSI [17 ]. P. aeruginosa ATCC 27853 was used as the quality control strain. The interpretation of resistance to these antimicrobial agents was determined according to the recommendations of the CLSI [18 ]. Multidrug resistant (MDR) P. aeruginosa was defined as isolates that were resistant to at least 3 classes of the tested antimicrobial agents [19 (link)].
8
Antibiotic Resistance Profiles of MDR Clinical Strains
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
This study included 31 MDR clinical strains carrying blaNDM and other associated resistance markers to determine the minimum inhibitory concentration (MIC) and fractional inhibitory concentration index (FICI). These strains were obtained from NICU of a North Indian Hospital. These are ESBL- and MBL-producing strains with different resistant markers, as reported previously by our group [7 (link),8 (link),9 (link),10 (link),11 (link),12 (link),13 (link)]. The antibiotic resistance markers and MIC of these strains are presented in Table 1 . Doripenem, cefoxitin, and imipenem were purchased from Sigma-Aldrich (Sigma, Milan, Italy). Streptomycin was purchased from Himedia (Mumbai, India). Mueller–Hinton broth was purchased from Himedia (Mumbai, India).
9
Doripenem and TGA-CdS QDs Synthesis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Doripenem (DOR, purity 99.9%) was purchased from Sigma Aldrich Chemie GmbH (Steinheim, Germany). Doripax® 500 mg vials were obtained from the local market of Assuit, Egypt. Cadmium chloride (CdCl2), sodium hydroxide (NaOH) and sodium sulfide (Na2S) were obtained from EL-Nasr Company (Egypt). Rhodamine B was obtained from Acros Organics (USA). Thioglycolic acid (TGA) was purchased from Sigma Aldrich Chemie GmbH (Steinheim, Germany).
A stock solution of DOR (1.0 mg mL−1) was prepared by precisely weighing 10.0 mg of DOR, transferring it into 10 mL volumetric flask and dissolving it using distilled water.
TGA-CdS QDs was synthesized following the method of Wang et al.,17 (link) the synthesis procedure and preparation of different reagent solutions are described in details in the ESI.† All the experiments included in this study followed the Assiut University and Al-Azhar University guidelines.
A stock solution of DOR (1.0 mg mL−1) was prepared by precisely weighing 10.0 mg of DOR, transferring it into 10 mL volumetric flask and dissolving it using distilled water.
TGA-CdS QDs was synthesized following the method of Wang et al.,17 (link) the synthesis procedure and preparation of different reagent solutions are described in details in the ESI.
10
Antimicrobial Susceptibility Testing Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Antimicrobial susceptibility was determined using a broth dilution method according to the Clinical and Laboratory Standards Institute (CLSI) guideline (CLSI, 2020 ). The minimum inhibitory concentrations (MICs) of colistin (Sigma), chloramphenicol (Duchefa), streptomycin (Sigma), gentamycin (Sigma), piperacillin (Duchefa), cefotaxime (Sigma), tetracycline (Sigma), ciprofloxacin (Sigma), doripenem (Sigma), imipenem (Sigma), meropenem (Sigma, Saint Louis, MO, USA) were determined using E. coli ATCC 25922 (American Type Culture Collection; ATCC, Manassas, VA, USA) as a quality control strain (CLSI, 2020 ). The antibiotic resistance level of chloramphenicol and streptomycin was determined based on previous studies (Wei and Yang, 2017 (link); Yang et al., 2019 (link)).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!