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1.
Water dispersible zinc sulfide quantum dots (ZnS QDs) with an average diameter of 2.9 nm were synthesized in an environment friendly method using chitosan as stabilizing agent. These nanocrystals displayed characteristic absorption and emission spectra having an absorbance edge at 300 nm and emission maxima (λ emission) at 427 nm. Citrate-capped silver nanoparticles (Ag NPs) of ca. 37-nm diameter were prepared by modified Turkevich process. The fluorescence of ZnS QDs was significantly quenched in presence of Ag NPs in a concentration-dependent manner with K sv value of 9 × 109 M−1. The quenching mechanism was analyzed using Stern–Volmer plot which indicated mixed nature of quenching. Static mechanism was evident from the formation of electrostatic complex between positively charged ZnS QDs and negatively charged Ag NPs as confirmed by absorbance study. Due to excellent overlap between ZnS QDs emission and surface plasmon resonance band of Ag NPs, the role of energy transfer process as an additional quenching mechanism was investigated by time-resolved fluorescence measurements. Time-correlated single-photon counting study demonstrated decrease in average lifetime of ZnS QDs fluorescence in presence of Ag NPs. The corresponding F?rster distance for the present QD–NP pair was calculated to be 18.4 nm.  相似文献   

2.
The photodynamic effects of deuteroporphyrin (DT) on the growth and viability ofStaphylococcus aureus, Streptococcus faecalis, andBacillus cereus are described. By exposure to light and DT, the growth ofSta. aureus andStr. Faecalis strains was markedly inhibited, whereasB. cereus was undergoing lysis. Counting the viable bacteria during the DT treatment revealed that more than 99% of the initial bacterial population was killed within the first 30 min of treatment. The pH dependence of the photodynamic inhibitory effect shows that it is best obtained at pH 6.5. No temperature dependence in the range of 37°–44°C could be detected. The best photodynamic effect was achieved when the culture was in the mid-log phase. DT-treated bacteria, when grown in the dark or in the presence of horse serum albumin, were unaffected by the porphyrin action. Electron microscopic examinations ofSta. aureus andStr. faecalis showed the appearance of well-developed mesosomes within the cell cytoplasm.B. cereus preparations have not shown any mesosome formation but showed some lysed cells as well as some spores. None of the mentioned effects by DT and light could be observed on Gramnegative bacteria such asEscherchia coli orPseudomonas aeruginosa.  相似文献   

3.
Generally, limited research is extended in studying stability and applicational properties of silver nanoparticles (Ag NPs) synthesized by adopting ‘green chemistry’ protocol. In this work, we report on the synthesis of stable Ag NPs using plant-derived materials such as leaf extract of Neem (Azadirachta indica) and biopolymer pectin from apple peel. In addition, the applicational properties of Ag NPs such as surface-enhanced Raman scattering (SERS) and antibacterial efficiencies were also investigated. As-synthesized nanoparticles (NPs) were characterized using various instrumentation techniques. Both the plant materials (leaf extract and biopolymer) favored the synthesis of well-defined NPs capped with biomaterials. The NPs were spherical in shape with an average particle size between 14-27 nm. These bio-NPs exhibited colloidal stability in most of the suspended solutions such as water, electrolyte solutions (NaCl; NaNO3), biological solution (bovine serum albumin), and in different pH solutions (pH 7; 9) for a reasonable time period of 120 hrs. Both the bio-NPs were observed to be SERS active through displaying intrinsic SERS signals of the Raman probe molecule (Nile blue A). The NPs were effective against the Escherichia coli bacterium when tested in nutrient broth and agar medium. Scanning and high-resolution transmission electron microscopy (SEM and HRTEM) images confirmed cellular membrane damage of nanoparticle treated E. coli cells. These environmental friendly template Ag NPs can be used as an antimicrobial agent and also for SERS based analytical applications.  相似文献   

4.
As the fundamental understanding of metal–light interactions gains solid grounds, further research has been devoted to construct novel structures that take full advantage of such unique interactions, which is called plasmonics. In this report, the preparation of Au–Ag core–shell structures obtained by coating the Au surface with peptide and Raman reporter molecule and depositing an Ag layer on it is reported. The prepared Au–Ag NPs are tested for their surface-enhanced Raman scattering (SERS) performance. The negatively charged peptides with three different lengths, which are 3 (P1), 15 (P2), and 21 (P3) amino acid long, were chemically attached to 13 nm AuNPs along with Raman reporter molecule, carboxytetramethylrhodamine, and these modified AuNPs were coated with three different shell thickness of Ag metal. The prepared Au–Ag NPs were tested for their SERS performance and found that the Au–Ag NPs prepared with P2 and thickest shell performs best as SERS label.  相似文献   

5.
The detection of biomarker-targeting surface-enhanced Raman scattering (SERS) nanoparticles (NPs) in the human gastrointestinal tract has the potential to improve early cancer detection; however, a clinically relevant device with rapid Raman-imaging capability has not been described. Here we report the design and in vivo demonstration of a miniature, non-contact, opto-electro-mechanical Raman device as an accessory to clinical endoscopes that can provide multiplexed molecular data via a panel of SERS NPs. This device enables rapid circumferential scanning of topologically complex luminal surfaces of hollow organs (e.g., colon and esophagus) and produces quantitative images of the relative concentrations of SERS NPs that are present. Human and swine studies have demonstrated the speed and simplicity of this technique. This approach also offers unparalleled multiplexing capabilities by simultaneously detecting the unique spectral fingerprints of multiple SERS NPs. Therefore, this new screening strategy has the potential to improve diagnosis and to guide therapy by enabling sensitive quantitative molecular detection of small and otherwise hard-to-detect lesions in the context of white-light endoscopy.  相似文献   

6.
Development of rapid and sensitive methods to detect pathogens is important to food and water safety. This study aimed to detect and discriminate important food- and waterborne bacteria (i.e., Escherichia coli O157:H7, Staphylococcus epidermidis, Listeria monocytogenes, and Enterococcus faecelis) by surface-enhanced Raman spectroscopy (SERS) coupled with intracellular nanosilver as SERS substrates. An in vivo molecular probing using intracellular nanosilver for the preparation of bacterial samples was established and assessed. Satisfactory SERS performance and characteristic SERS spectra were obtained from different bacterial samples. Distinctive differences were observed in SERS spectral data, specifically in the Raman shift region of 500–1,800 cm−1, and between bacterial samples at the species and strain levels. The detection limit of SERS coupled with in vivo molecular probing using silver nanosubstrates could reach the level of single cells. Experiments with a mixture of E. coli O157:H7 and S. epidermidis for SERS measurement demonstrate that SERS could be used for classification of mixed bacterial samples. Transmission electron microscopy was used to characterize changes of morphology and cellular composition of bacterial cells after treatment of intracellular nanosilver. The results indicate that SERS coupled with intracellular silver nanosubstrates is a promising method for detection and characterization of food- and waterborne pathogenic and non-pathogenic bacterial samples.  相似文献   

7.
A new type of nitrogen and chloride co-doped carbon dots (N/Cl-CDs) based on choline chloride–urea–glycine ternary deep eutectic solvents (DESs) was synthesized using a one-step hydrothermal method. The prepared N/Cl-CDs exhibited oxidase-like activity and excellent antibacterial activity against Escherichia coli, Staphylococcus aureus and methicillin-resistant Staphylococcus aureus (MRSA). The addition of silver nanoparticles (Ag NPs) (i.e. N/Cl-CDs + Ag NPs) to the N/Cl-CDs also significantly enhanced the oxidase and antibacterial activities. The nanocomposite (1·8 mg ml−1) completely inactivated 105 CFU per ml of MRSA in 90 min. E. coli and S. aureus were labelled with the N/Cl-CDs, enabling multicolour fluorescence imaging at different excitation wavelengths. The nanocomposites have high antibacterial efficiency as a new bactericidal agent, as well as application potential with good biocompatibility and low toxicity.  相似文献   

8.
Staphylococcus aureus, particularly methicillin-resistant S. aureus (MRSA), is an important cause of pyogenic skin and soft tissue infections (SSTIs). MRSA is an important pathogen in the healthcare sector that has neither been eliminated from the hospital nor community environment. In humans, S. aureus causes superficial lesions in the skin and localized abscesses, pyogenic meningitis/encephalitis, osteomyelitis, septic arthritis, invasive endocarditis, pneumonia, urinary tract infections and septicemia. Investigations focused in the search of other alternatives for the treatment of MRSA infections are in progress. Among the range of compounds whose bactericidal activity is being investigated, ZnO nanoparticles (ZnO–NPs) appears most promising new unconventional antibacterial agent that could be helpful to confront this and other drug-resistant bacteria. The aim of present study is to investigate the antibacterial potential of ZnO–NPs against Staphylococcus species isolated from the pus and wounds swab from the patients with skin and soft tissue infections in a tertiary care hospital of north India. ZnO–NPs (≈19.82 nm) synthesized by sol–gel process were characterized using scanning electron microscopy, X-ray diffraction , and Atomic force microscopy. The antibacterial potential was assessed using time-dependent growth inhibition assay, well diffusion test, MIC and MBC test and colony forming units methods. ZnO–NPs inhibited bacterial growth of methicillin-sensitive S. aureus (MSSA), MRSA and methicillin-resistant S. epidermidis (MRSE) strains and were effective bactericidal agents that were not affected by drug-resistant mechanisms of MRSA and MRSE.  相似文献   

9.
Matrix-assisted laser desorption-ionization (MALDI) time-of-flight mass spectrometry was used to characterize the spores of 14 microorganisms of the Bacillus cereus group. This group includes the four Bacillus species B. anthracis, B. cereus, B. mycoides, and B. thuringiensis. MALDI mass spectra obtained from whole bacterial spores showed many similarities between the species, except for B. mycoides. At the same time, unique mass spectra could be obtained for the different B. cereus and B. thuringiensis strains, allowing for differentiation at the strain level. To increase the number of detectable biomarkers in the usually peak-poor MALDI spectra of spores, the spores were treated by corona plasma discharge (CPD) or sonicated prior to MALDI analysis. Spectra of sonicated or CPD-treated spores displayed an ensemble of biomarkers common for B. cereus group bacteria. Based on the spectra available, these biomarkers differentiate B. cereus group spores from those of Bacillus subtilis and Bacillus globigii. The effect of growth medium on MALDI spectra of spores was also explored.  相似文献   

10.
The target of our current work was designed to prepare titanium oxide doped silver nanoparticles (Ag/TiO2NPs) and their impact on the functionalization of cotton fabrics. Additionally, the effect of Ag/TiO2NPs was compared with the individually prepared silver nanoparticles (AgNPs) and titanium oxide nanoparticles (TiO2NPs). In this work, AgNPs were prepared in the solid state using arabic gum as efficient stabilizing and reducing agent. Then, two concentrations of the as-synthesized nanoparticles were used to functionalize the cotton fabrics by pad-dry-cure treatment in the presence of fixing agent to increase the durability of treated cotton fabrics against vigorous washing cycles. The findings implied that the as-prepared nanoparticles were successfully synthesized in nano-size with spherical shape and homogeneity. The efficacy of the functionalized cotton fabrics with those nanoparticles were evaluated in terms of multifunctional properties including antimicrobial and ultraviolet protection factor (UPF) and the mechanical features before and after many washing cycles; 10, 15 and 20 times. The resultant also proved that Ag/TiO2NPs-treated cotton fabrics exhibited the greater values of both antimicrobial and UPF properties with enhancement in the tensile strength and elongation features. Thus, the combination between these two nanoparticles through doping reaction is suitable for imparting superior antimicrobial properties against the four tested microbial species (Staphylococcus aureus, Escherichia coli, Candida albicans, and Aspergillus niger) and good UPF properties. Depending on the promising obtained results of the multi-finishing fabrics, these nanoparticles of Ag/TiO2NPs can be applied for the production of an efficient medical clothes for doctors, nurses and bed sheets for patients in order to kill and prevent the spread of bacteria and then, reduce the transmission of infection to others.  相似文献   

11.
Teng  Yuanjie  Wang  Zhenni  Ren  Zeyu  Qin  Yanping  Pan  Zaifa  Shao  Kang  She  Yuanbin  Huang  Weihao 《Plasmonics (Norwell, Mass.)》2021,16(2):349-358

Water-insoluble molecules usually show poor surface-enhanced Raman scattering (SERS) signals, because they are hardly adsorbed on the surface of most commonly used SERS substrates, such as aqueous Ag or Au colloids. In this work, a highly sensitive and reproducible Ag monolayer film (Ag MLF) SERS substrate prepared by self-assembly of Ag nanoparticles (Ag NPs) on water/oil interface can realize the trace SERS detection of water-insoluble enrofloxacin. The positively charged phase transfer catalyst can transfer the negatively charged Ag nanoparticles in aqueous solution to the water/oil interface. At the same time, the water-insoluble enrofloxacin can also be attracted to the interface because of its lipophilic group. The type/volume of the oil phase and phase transfer catalyst and the vortex mixing time were all optimized to maximize the SERS effect of Ag MLF. Results showed that trace water-insoluble enrofloxacin can be identified by Ag MLF and its detection sensitivity was significantly improved. The proposed novel Ag MLF can be further applied to detect other water-insoluble molecules in SERS.

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12.
Highly resistant pathogens may be developed in patients with immune disorders after prolonged exposure to antibiotics, a growing threat worldwide. In order to overcome these problems, this study introduces a new class of engineered nanosystems comprising of tea tree oil nanoemulsion (TTO NE) loaded with Ag nanoparticles (NPs). Silver shows a strong toxicity towards a wide range of microorganisms. Also, TTO NE could be employed as a promising and safe antimicrobial agent for local therapies of bacterial infections. The nanosystem was prepared by low-energy method. Mean droplet size of the NE was found to be 17.7 nm. Results of the antibacterial assays showed promising ability of the designed nanosystem for eradication of Gram-positive and Gram-negative bacteria (95%). Also, it was shown that introducing colloidal Ag NPs to the TTO NE exerted a synergistic effect against Escherichia coli (FIC 0.48) while only an additive effect was observed against Staphylococcus aureus (FIC 0.75). The antibacterial effects of TTO NE+Ag NPs together with their compatibility with human cells can present them as a suitable candidate to fight against the antibacterial resistance threat.  相似文献   

13.
The ability of noble metal‐based nanoparticles (NPs) (Au, Ag) to drastically enhance Raman scattering from molecules placed near metal surface, termed as surface‐enhanced Raman scattering (SERS), is widely used for identification of trace amounts of biological materials in biomedical, food safety and security applications. However, conventional NPs synthesized by colloidal chemistry are typically contaminated by nonbiocompatible by‐products (surfactants, anions), which can have negative impacts on many live objects under examination (cells, bacteria) and thus decrease the precision of bioidentification. In this article, we explore novel ultrapure laser‐synthesized Au‐based nanomaterials, including Au NPs and AuSi hybrid nanostructures, as mobile SERS probes in tasks of bacteria detection. We show that these Au‐based nanomaterials can efficiently enhance Raman signals from model R6G molecules, while the enhancement factor depends on the content of Au in NP composition. Profiting from the observed enhancement and purity of laser‐synthesized nanomaterials, we demonstrate successful identification of 2 types of bacteria (Listeria innocua and Escherichia coli). The obtained results promise less disturbing studies of biological systems based on good biocompatibility of contamination‐free laser‐synthesized nanomaterials.

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14.
3-Allyl-5-(4-arylazo)-2-thioxothiazolidine-4-one (HLn) ligands (where n = 1 to 3) were hypothesized to have antimicrobial activities mediated through inhibition of new antimicrobial targets. The ligands (HLn) were synthesized and characterized by infrared (IR) and 1H nuclear magnetic resonance (1H NMR) spectra. The ligands (HLn) were in silico screened to their potential inhibition to models of d -alanyl carrier protein ligase (DltA) (from Bacillus cereus, PDB code 3FCE) and nucleoside diphosphate kinase (NDK) (from Staphylococcus aureus; PDB code 3Q8U). HL3 ligand has the best energy and mode of binding to both NDK and DltA, even though its binding to DltA was stronger than that to NDK. In antimicrobial activity of HL3 ligand, morphological and cytological changes in HL3-treated bacteria agreed with the in silico results. The HL3 ligand showed significant antimicrobial activity against B. cereus, S. aureus, and Fusarium oxysporium. The HL3-treated bacterial cells appeared malformed and incompletely separated. Its cell walls appeared electron-lucent and ruptured. They contained more mesosomes than normal cells. It was found that the HL3 ligand represented as a bactericide against B. cereus and S. aureusby blocking target DltA, and may target NDK.  相似文献   

15.

Silver (Ag) nanoparticles (NPs) and Ag nanorings (NRs) have been fabricated. Due to the inherent features of Ag NPs and Ag NRs, strong electromagnetic (EM) near-field distributions were expected, and hence surface-enhanced Raman scattering (SERS) activity was demonstrated. Size and interparticle gaps distribution of Ag NPs were estimated to be 48.14?±?10.14 nm and 14.11?±?5.24 nm respectively along with estimated coverage density of?~?4?×?1010 cm?2. On the other hand, Ag NRs were found to consist of Ag clusters and of various shapes and sizes, instead of a perfect ring structure. High-resolution FESEM revealed that the individual constituent clusters were different from each other, particularly in terms of size and shape in addition to the cases how such clusters were connected to form the edge of the NR. However, the coverage density of Ag NRs was estimated to be?~?5.6?×?106 cm?2. Based on the scenarios, it was speculated that the local EM near-field distribution would excel and thus led to enhanced SERS signals. SERS enhancement of R6G was estimated as high as 2.18?×?104 and 2.78?×?104 at 610 cm?1 (C???C ring bending mode in phenyl rings) for Ag NPs and Ag NRs respectively. FDTD analysis was carried out to elucidate the EM near-field distributions.

Graphical abstract

Ag NPs and Ag NRs from an ultrathin layer of Ag on ZnO/Glass (middle pane) confirming high EF of R6G adsorbed on Ag NRs (right pane) and Ag NPs (left pane) supported by corresponding EM near-field distributions.

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16.
Highly stable silver nanoparticles (Ag NPs) in agar-agar (Ag/agar) as inorganic-organic hybrid were obtained as free-standing film by in situ reduction of silver nitrate by ethanol. The antimicrobial activity of Ag/agar film on Escherichia coli (E. coli), Staphylococcus aureus (S. aureus), and Candida albicans (C. albicans) was evaluated in a nutrient broth and also in saline solution. In particular, films were repeatedly tested for antimicrobial activity after recycling. UV-vis absorption and TEM studies were carried out on films at different stages and morphological studies on microbes were carried out by SEM. Results showed spherical Ag NPs of size 15-25 nm, having sharp surface plasmon resonance (SPR) band. The antimicrobial activity of Ag/agar film was found to be in the order, C. albicans > E. coli > S. aureus, and antimicrobial activity against C. albicans was almost maintained even after the third cycle. Whereas, in case of E. coli and S. aureus there was a sharp decline in antimicrobial activity after the second cycle. Agglomeration of Ag NPs in Ag/agar film on exposure to microbes was observed by TEM studies. Cytotoxic experiments carried out on HeLa cells showed a threshold Ag NPs concentration of 60 μg/mL, much higher than the minimum inhibition concentration of Ag NPs (25.8 μg/mL) for E. coli. The mechanical strength of the film determined by nanoindentation technique showed almost retention of the strength even after repeated cycle.  相似文献   

17.
In the current investigation, we report the biosynthesis of silver nanoparticles (Ag NPs) employing extract of Alternaria alternata, which is an eco-friendly process for the synthesis of metallic nanoparticles. Ag NPs were synthesised through the reduction of aqueous Ag+ ion using the cell extract of fungus A. alternata in the dark conditions. The synthetic process was relatively fast and Ag NPs were formed within 24 h. UV–visible spectrum of the aqueous medium containing silver ion showed a peak at 435?nm corresponding to the plasmon absorbance of Ag NPs and another peak at 280?nm refers to tyrosine amino acid. The nanoparticles were characterised by scanning electron microscopy (SEM) and Fourier transform infrared spectroscopy (FTIR). The morphology of nanoparticles is found to be spherical mostly, with ranging size of 27–79?nm; as revealed by SEM. The FTIR spectrum analysis indicated that biomolecules were involved in the synthesis of Ag NPs. The presence of the amino groups is expected to pack differently around the Ag NPs. This in turn will influence the self-assembly of nanoparticles on substrates as well as their stability. The present study demonstrates the possible use of biologically synthesised Ag NPs in the field of agriculture, when A. alternata could be used for simple, nonhazardous and efficient synthesis of Ag NPs.  相似文献   

18.
New biochip technology for label-free detection of pathogens and their toxins   总被引:10,自引:0,他引:10  
microSERS is a new biochip technology that uses surface-enhanced Raman scattering (SERS) microscopy for label-free transduction. The biochip itself comprises pixels of capture biomolecules immobilized on a SERS-active metal surface. Once the biochip has been exposed to the sample and the capture biomolecules have selectively bound their ligands, a Raman microscope is used to collect SERS fingerprints from the pixels on the chip. SERS, like other whole-organism fingerprinting techniques, is very specific. Our initial studies have shown that the Gram-positive Listeria and Gram-negative Legionella bacteria, Bacillus spores and Cryptosporidium oocysts can often be identified at the subspecies/strain level on the basis of SERS fingerprints collected from single organisms. Therefore, pathogens can be individually identified by microSERS, even when organisms that cross-react with the capture biomolecules are present in a sample. Moreover, the SERS fingerprint reflects the physiological state of a bacterial cell, e.g., when pathogenic Listeria and Legionella were cultured under conditions known to affect virulence, their SERS fingerprints changed significantly. Similarly, nonviable (e.g., heat- or UV-killed) microorganisms could be differentiated from their viable counterparts by SERS fingerprinting. Finally, microSERS is also capable of the sensitive and highly specific detection of toxins. Toxins that comprised as little as 0.02% by weight of the biomolecule-toxin complex produced strong, unique fingerprints when spectra collected from the complexes were subtracted from the spectra of the uncomplexed biomolecules. For example, aflatoxins B(1) and G(1) could be detected and individually identified when biochips bearing pixels of antibody or enzyme capture biomolecules were incubated in samples containing one or both aflatoxins, and the spectra were then collected for 20 s from an area of the biomolecule pixel approximately 1 microm in diameter. In the future, we plan to investigate the use of hyperspectral imaging Raman microscopy for collecting fingerprints from all the pixels on the biochip, individually yet simultaneously, to enable the rapid detection of diverse pathogens and their toxins in a sample, using a single biochip.  相似文献   

19.
In the present report, Aegle marmelos leaf powder was used to synthesize copper nanoparticles (CuNPs) using a simple and cost-effective method. A. marmelos leaves have various medicinal uses including for the treatment of diarrhoea, constipation, diabetes, cholera, skin diseases, earache, blood purification, heart problems, and so on. The plant biomolecules induce the reduction of Cu2+ ions to CuNPs and also act as a capping and stabilizing agent. The formation of CuNPs was confirmed using photoluminescence (PL) excitation and emission spectra on a Shimadzu RF-5301 PC spectrofluorophotometer and the absorbance spectra of a UV–visible spectrophotometer at different stages during the synthesis process. In addition, other properties of synthesized CuNPs were also investigated using X-ray diffraction, Fourier transform infrared spectroscopy, and transmission electron microscopy techniques. The average size of the synthesized CuNPs was in the range 20–40 nm. Furthermore, the synthesized NPs were also considered for an antimicrobial study against Gram-positive Staphylococcus aureus and Proteus, and Gram-negative Escherichia coli and Salmonella spp. using the agar well diffusion method. The zone of inhibition against the Gram-positive bacteria was greater than the zone of inhibition against the Gram-negative bacteria. These investigation results suggest that synthesized NPs are promising nanomaterials for use as antimicrobial agents.  相似文献   

20.
It was shown that matrix-assisted laser desorption/ionization (MALDI) mass spectrometry could be used for the diagnostic characterization of propionic acid bacteria (PABs). The spectra of proteins (whole PAB cells) with a molecular mass of 3000 to 11 000 were obtained and analyzed using three matrices: sinapinic (SA), 2,5-dihydroxibenzoic (DHB), and α-cyano-4-hydroxycinnamic acid (HCCA). The MALDI spectra of PAB revealed the protein peaks characteristic of (1) the genus Propionibacterium (3496, 5386, 5605, 10 470), (2) the groups of species sharing the common composition of their cell walls and fatty acids, and (3) a species (four species were investigated). Exemplified by the P. shermanii strains (the collection and mutant ones) producing and not producing vitamin B12, the possibility of using MALDI profiles for strain differentiation was confirmed. The MALDI profiles of the propionic acid cocci of the genus Luteococcus differ substantially from the profiles of PAB strains of the genus Propionibacterium, which is an additional proof of the validity of whole-cell MALDI spectra for generic differentiation of bacteria. Our investigation shows that the bacterial groups determined using the MALDI profiles correlate with the phylogenetic 16S rRNA gene groups, thus demonstrating the high resolution of this method for the differentiation of intraspecific differences (subspecies, strains).  相似文献   

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