Kanamycin Research Articles

Voltammetric kanamycin aptasensor based on the use of thionine incorporated into Au@Pt core-shell nanoparticles.

A signal-on aptasensor is described for the voltammetric determination of kanamycin (KANA). Au@Pt core-shell nanoparticles with large surface and good electrical conductivity were synthetized and act as both a conductive material and as the carrier for complementary strands (CS2) and thionine (TH). In the presence of KANA, the electrochemical response of TH changes due to hybridization between CS1 immobilized on the electrode and the Au@Pt-CS2/TH system. The peak current increases linearly with the logarithm of the KANA concentration in the range from 1 pM to 1μM, and the limit of detection is 0.16 pM. The sensor was characterized in terms of selectivity, reproducibility and stability, and satisfactory results were obtained. It was also utilized for the determination of KANA in (spiked) chicken samples. The recoveries (95.8-103.2%) demonstrate the potential of the method for KANA detection in real samples. Graphical abstract A signal-on aptasensor for kanamycin (KANA) was developed by using Au@Pt core-shell nanoparticles as nanocarrier for probe aptamer and as asensing probe.

Ultrasensitive analysis of kanamycin residue in milk by SERS-based aptasensor

An ultrasensitive method for the kanamycin (KANA) detection in milk sample using surface-enhanced Raman spectroscopy-based aptasensor was employed in the current study. Double strand DNA binding bimetallic gold@silver nanoparticles were developed as a sensing platform. Probe DNAs were first embedded on the surface of gold nanoparticles by the end-modified thiol, and after silver shell encapsulating, KANA aptamer DNAs with the Raman reporter Cy3 were then hybridized with probe DNAs by complementary base pairing. Results showed that with increase in the KANA concentration, the Raman intensity of Cy3 decreased. Besides achieving selectivity, an ultralow detection limit of 0.90 pg/mL, a broad linear relationship ranging from 10 μg/mL to 100 ng/mL in aqueous reagent and satisfactory recoveries of 90.4–112% in liquid whole milk were obtained. The result of actual sample proved that this aptasensor was promising in trace determination of KANA residue.

Construction of Tn5 transposon insertion mutants of Ralstonia solanacearum isolated from Pogostemon cablin

Ralstonia solanacearum strain PRS-84 used in this study was isolated from diseased Pogostemon cablin plants in our previous study.The competent cells of R.solanacearum strain PRS-84 were transformed by electroporation with Tn5 transposon and then were plated on TTC agar plates containing kanamycin to select for kanamycin-resistant colonies.The detection of kanamycin-resistant gene in kanamycin-resistant colonies was performed by PCR.Further,the flanking fragments of Tn5 transposon insertion site in the mutants were amplified by inverse PCR,and the flanking fragments were sequenced and analyzed.The results indicated that the kanamycin-resistant colonies were obtained in the transformation experiment of R.solanacearum strain PRS-84 by electroporation with Tn5 transposon.A specific band of approximately 700 bp was amplified by PCR from kanamycin-resistant colonies.The flanking sequences of Tn5 transposon insertion site in the transformants were obtained by inverse PCR.After sequencing and sequence analysis of Tn5 transposon insertion site in mutants,we preliminarily speculated that the Tn5 transposon inserted in the typ A gene,rec O gene and gid A gene in three mutants,respectively.A random mutagenesis system of R.solanacearum strain PRS-84 by electroporation with Tn5 transposon has been established,and the Tn5 insertion mutants have been obtained.This study might facilitate the creation of mutant library and the discovery of the virulence gene of R.solanacearum isolated from P.cablin.

A novel fluorescent sensing platform based on metal-polydopamine frameworks for the dual detection of kanamycin and oxytetracycline.

In this study, we report the dual detection of kanamycin (KMY) and oxytetracycline (OTC) using metal polydopamine frameworks (MPDA) for the first time. This sensing system employed metal polydopamine frameworks (MPDA), which acted as a fluorescence quencher in the interaction between MPDA and dye-labeled DNA molecules; the metal polydopamine frameworks exhibited an excellent fluorescence quenching behaviour and good analytical response towards the detection of the biomolecules KMY and OTC. The accumulated kanamycin and oxytetracycline in the sensing system were recovered, and changes in their fluorescence intensity were monitored at 525 and 583 nm. Under the optimal conditions, the proposed sensing system remarkably achieved highly sensitive and selective detection of KMY and OTC with the limit of detection of 304 pM and 481 pM, respectively. In addition, the selectivity of the developed sensor was explored in the presence of competitive biomolecules. Moreover, this sensing system demonstrated great potential and versatility for the rapid detection of molecules. In addition, this biosensor was successfully evaluated for the dual detection of KMY and OTC in different real samples.

Detection of 16S rRNA gene and biological characteristics of Mycobacterium abscessus

Objective To identify a clinical isolate of Mycobacterium abscessus. Methods A pus sample was collected from a patient with suspected nontuberculous mycobacterial infection who visited the Affiliated Hospital of Weifang Medical University on December 18, 2017, and was subjected to bacterial culture, Gram staining and acid-fast staining. Drug sensitivity test was conducted by the proportion method. The genome DNA of the strain was extracted and amplified by PCR with the universal primer of 16S rRNA. The PCR products were sequenced after collection and purification, and were compared with the known sequence of Mycobacterium abscessus in GenBank database. The isolate was identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) . Results The clinical isolate was identified as Mycobacterium abscessus both by MALDI-TOF MS and 16S rRNA gene sequencing. The drug sensitivity test showed that the strain was sensitive to amikacin, moxifloxacin, levofloxacin, but was resistant to streptomycin, isoniazid, rifampicin, ethambutol, ofloxacin, kanamycin, capreomycin, aminosalicylic acid, protionamide and rifabutin. The patient was diagnosed with subcutaneous soft tissue infection in the left knee joint. According to the results of drug sensitivity test, the patient was treated with amikacin and levofloxacin, and her condition was improved after treatment. Conclusion The 16S rRNA gene detection and MALDI-TOF MS both can be applied in the identification of Mycobacterium abscessus. Key words: Nontuberculous mycobacteria; RNA, ribosomal, 16S; Spectrometry, mass, matrix-assisted laser desorption-ionization; Microbial sensitivity tests; Mycobacterium abscessus

Multiplexed electrochemical aptasensor for antibiotics detection using metallic-encoded apoferritin probes and double stirring bars-assisted target recycling for signal amplification

Simultaneous and sensitive detection of various antibiotic residues in one sample is essential to evaluation of food safety status. Herein, a multiplexed electrochemical aptasensor for multiplex antibiotics detection, with kanamycin (KANA) and ampicillin (AMP) as representative analytes, was designed by using metal ions encoded apoferrtin probes and double stirring bars-assisted target recycling for signal amplification. The encoded probes were prepared by apoferritin loading Cd2+ and Pb2+ ions and labeling with duplex DNAs (aptamers corresponding to KANA and AMP hybrid with its complementary DNA sequence), respectively. In the presence of KANA and AMP, the targets can recurrently react with the probes on the bars, and then replace a lot of Apo-Mencoded signal tags into supernatant. The peak currents of Cd2+and Pb2+from the tags corresponding with the concentrations of KANA and AMP were detected by square wave voltammetry in one run. As a result, KANA and AMP can be detected simultaneously within the range from 0.05 pM to 50 nM. And the detection limits were 18 fM KANA and 15 fM AMP (S/N = 3). The assay was testified to detect KANA and AMP residues with consistent results of ELISA in samples, e.g. milks and fishes. The assay was highly-sensitive, selective, cost-effective and easy-to-operate due to Apo-M encoded probes with high loading capacity of signal source substances. Moreover, double stirring bar-assisted target recycling, which was enzyme-free and could overcome matrix interference, was fabricated for signal amplification. Thus, the assay showed potential advantages for sensitively screening of antibiotic residues in foods.

Rapid Detection of AGs using Microchip Capillary Electrophoresis Contactless Conductivity Detection

Background: In order to realize current aminoglycosides supervision in food and environment, our team improved the sensitivity and separation efficiency of the portable ITO detector, based on the technology of microchip capillary electrophoresis and contactless conductivity detection. Experiment: Parameters (the separation voltage, buffer concentration, electrodes gap, elicitation frequency, elicitation voltage) were optimized for the detection of three aminoglycosides, gentamicin, kanamycin and streptomycin and the separation of their mixture in background electrolyte consists of 2-(N-Morpholino) ethanesulfonic acid (MES) and L-Histidine (His). The enhanced method was also applied to other types of aminoglycosides. Results: Under optimal conditions, the monitoring of three types of aminoglycosides obtained such a sensitive response that the limits of detection of gentamicin sulfate, kanamycin sulfate and streptomycin sulfate were calculated as 3.1 µg/ml, 0.89 µg/ml and 0.96 µg/ml, at signal-to-noise ratio 3, respectively. In addition they got separated completely from each other only in 40 s. The results of other varieties of aminoglycosides including tobramycin sulfate and amikacin sulfate also met the standard. Conclusion: We successfully proposed here an unprecedentedly portable, miniaturized and rapid microchip capillary electrophoresis contactless conductivity detection system to realize current aminoglycosides supervision in food and environment.

A colorimetric aptasensor for the antibiotics oxytetracycline and kanamycin based on the use of magnetic beads and gold nanoparticles.

An aptamer based assay is presented for the determination of the antibiotics oxytetracycline (OTC) and kanamycin (KAN). Magnetic beads were applied for separation, and gold nanoparticles (AuNPs) for signal amplification. DNA aptamers against OTC and KAN were firstly designed. After specific recognition events, the aptamer sequences were released from the surface of magnetic beads and the remaining DNA probes captured horseradish peroxidase (HRP) modified AuNPs. Subsequently, 3,3',5,5'-tetramethylbenzidine and o-phenylenediamine are catalytically oxidized by HRP, and the generated colorimetric responses can reflect the concentrations of OTC (at 370nm) and KAN (at 450nm), respectively. Experimental results demonstrate that the method is highly sensitive with the detection limit as low as 1 ag mL-1 for OTC and KAN. An extremely wide linear range (over 11 orders of magnitude) is achieved. The high selectivity is attributed to the high affinity between aptamer and the substrate. The results of real sample tests also verify that the method is promising for antibiotics analysis in the applications of food monitoring and clinical diagnosis. Graphical abstract Schematic presentation of a colorimetric assay for antibiotics based on aptamer-modified magnetic beads and horseradish peroxidase modified gold nanoparticles. Colorimetric responses result from the enzymatic oxidation of 3,3',5,5'-tetramethylbenzidine (TMB) and o-phenylenediamine (OPD), respectively.

Antibacterial and Antibiotic-Potentiating Activities of Thirteen Cameroonian Edible Plants against Gram-Negative Resistant Phenotypes.

This work was designed to investigate the antibacterial activities of methanol extracts from thirteen Cameroonian edible plants and their antibiotic-potentiating effects against Gram-negative multidrug-resistant (MDR) phenotypes. The broth microdilution method was used to evaluate the minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) of the extracts, as well as their antibiotic-potentiating activities. The phytochemical screening of the extracts was carried out according to the standard methods. The results of phytochemical tests revealed the presence of sterols, polyphenols, and tannins in most of the tested extracts, with the other classes of secondary metabolites being selectively distributed. Tested extracts showed variable antibacterial activities with MIC values ranging from 64 to 1024 μg/mL. However, some extracts were significantly active against certain bacterial strains: seeds extract of Theobroma cacao (64 μg/mL) against Escherichia coli AG100Atet and Klebsiella pneumoniae K24, and the bark extract of Uapaca guineensis against E. coli ATCC 8739. The leaves extract of T. cacao displayed the best MBC values (256 μg/mL) against E. aerogenes EA27. Some tested extracts included extracts from the leaves of T. cacao and P. vulgaris, and the seeds of D. edulis and barks A. indica has selectively improved (2- to 64-fold) the antibacterial activities of some of the tested antibiotics, chloramphenicol (CHL), tetracycline (TET), kanamycin (KAN), streptomycin (STR), and erythromycin (ERY), against more than 70% of tested MDR bacteria. The findings of this work showed that tested plant extracts and particularly those from T. cacao and Phaseolus vulgaris can be used alone or in combination with conventional antibiotics in the treatment of infections involving multiresistant bacteria.

A Chip for Detecting Tuberculosis Drug Resistance Based on Polymerase Chain Reaction (PCR)-Magnetic Bead Molecule Platform.

Objective: A Tag Array chip was used to detect plasmids of different template concentration, and then analyzed for sensitivity and specificity. Drug resistance genes from tuberculosis clinical specimens were detected, giving comparative phenotypic resistance results to explore the feasibility and value of clinical applications.Methods: Twenty-four strains of Mycobacterium Tuberculosis (MTB) having sequence differences in extracted plasmids of mutant strains. The plasmid was diluted into different concentrations, and then was performed to analyze the sensitivity and specificity of the chip system. A total of 427 clinical specimens (including spinal tuberculosis and pulmonary tuberculosis) were collected from patients who came from seven hospitals. Design, optimization and preparation of the chip detection system, sequencing and phenotypic drug susceptibility results were analyzed to evaluate the sensitivity and specificity of the gene chip.Results: In the template, concentrations of 1 × 103 copies/μL and above were consistent with sequencing results in the mutant. The sensitivity and specificity in spine Tuberculosis specimen of rifampicin (RFP) were 94.40 and 92.86%; isoniazide (INH) were 92.37 and 87.50%; ethambutol (EMB) were 61.36 and 89.29%; fluoroquinolones (FQS) were 79.41 and 92.86%; streptomycin (SM) were 90.18 and 89.29%; second line drugs (SLD) were 77.61 and 83.93%. In Pulmonary Tuberculosis specimen, the sensitivity and specificity respectively were RFP: 92.74%; 93.75%; INH: 91.26%; 87.50%; EMB: 54.17%; 89.58%; FQS: 84.87%; 93.75%; SM: 86.73%; 85.42%; SLD: 80.9%; 91.67%. The RFP, INH, FQs and SM resistance genes was highly sensitive and specific: however, for detection of amikacin (AMK), capreomycin (CPM), kanamycin (KM), specificity was higher, but sensitivity was lower. Sensitivity for the detection of a mutation in the eis promoter region could be improved.Conclusion: Tag Array chip can achieve rapid, accurate detection of tuberculosis resistance, which has important clinical significance and feasibility.

The Novel Peptide Vaccine GV1001 Protects Hearing in a Kanamycin-induced Ototoxicity Mouse Model.

We tested whether GV1001 has any ototoxic side effects at different doses and whether it protects hearing in an aminoglycoside-induced ototoxicity mouse model. GV1001, a novel peptide vaccine currently being examined in a Phase 3 clinical trial to treat pancreatic cancer, also has anti-inflammatory and antioxidant effects. In the first experiment, C57/BL6 mice were injected with GV1001 preparations at concentrations of 0.1 to 100 mg/kg for 7 days to evaluate the toxicity of GV1001 on the inner ear and kidneys. In the second experiment, the protective effect of GV1001 was tested in an ototoxicity mouse model that was generated by injecting 800 mg/kg kanamycin (KM) for 2 weeks. The hearing threshold and hair cell loss were compared between the KM + GV1001 group (treated with 10 mg/kg GV1001 for 2 wk) and the KM + saline group. The hearing threshold was measured before, and 7, 14, and 21 days after the initial treatment. The blood urea nitrogen level was measured. No ototoxicity or renal toxicity was found following treatment with different doses of GV1001 (0.1-100 mg/kg). The KM + saline group showed impaired auditory function and markedly disoriented and missing cochlear hair cells, while the KM + GV1001 group showed significant hearing and hair cell preservation in comparison (p < 0.05). GV1001 itself did not have any detrimental effects on the inner ear or kidney. In the KM induced ototoxicity model, concomitant administration of GV1001 protected against cochlear hair cell damage and preserve hearing.

Phenotypic characterization of indigenous rhizobia nodulating chickpea in Turkey reveals high diversity

A total of 120 root-nodule bacteria from chickpea plants (Cicer arietinum L.) from six regions in Turkey were characterized using 71 phenotypes. Utilization of carbon and nitrogen sources, tolerance to salt stress, temperature and pH, resistance to antibiotics and heavy metals and ability to produce some enzymes, were assessed. 90% of the isolates produced mucously, circular, smooth-margined and watery to creamy colonies with 2–4 mm diameter after 1-3 days of growth. The isolates utilized different compounds as sole carbon and nitrogen sources, endured 2% salt, grew optimally at 25-35°C and pHs between 6-8, exhibited insensitivity to the heavy metals Zn, Hg, and Cu and the antibiotics kanamycin, streptomycin and tetracycline. Numerical analysis separated the isolates into three clusters at 25% similarity. Results revealed diversity among isolates and were consistent with previous findings on chickpea Mesorhizobia.

Effect of lidocaine on kanamycin injection-site pain in patients with multidrug-resistant tuberculosis.

SETTING:Reducing pain from intramuscular injection of kanamycin (KM) could improve the tolerability of multidrug-resistant tuberculosis (MDR-TB) treatment. Lidocaine has been shown to be an effective anaesthetic diluent for some intramuscular injections, but has not been investigated with KM in the treatment of adult patients with MDR-TB.OBJECTIVE AND DESIGN:We performed a randomised single-blinded crossover study to determine if lidocaine reduces KM injection-site pain. We recruited patients aged ⩾18 years on MDR-TB treatment at two TB hospitals in Cape Town, South Africa. KM pharmacokinetic parameters and a validated numeric pain scale were used at intervals over 10 h following the injection of KM with and without lidocaine on two separate occasions.RESULTS:Twenty participants completed the study: 11 were males, the median age was 36 years, 11 were HIV-infected, and the median body mass index was 17.5 kg/m2. The highest pain scores occurred early, and the median pain score was 0 by 30 min. The use of lidocaine with KM significantly reduced pain at the time of injection and 15 min post-dose. On multiple regression analysis, lidocaine halved pain scores (adjusted OR 0.5, 95%CI 0.3–0.9). The area under the curve at 0–10 h of KM with and without lidocaine was respectively 147.7 and 143.6 μg·h/ml.CONCLUSION:Lidocaine significantly reduces early injection-site pain and has no effect on KM pharmacokinetics.

A microchip electrophoresis-based assay for ratiometric detection of kanamycin by R-shape probe and exonuclease-assisted signal amplification

Excessive intake of kanamycin (KANA) can cause some serious drug-resistant diseases, so it is urgent to develop some accurate and rapid analytical methods for monitor KANA residues in foodstuffs with complex matrix. Recently, many ratiometric assays were reported to be capable of overcoming matrix interference. Herein, a ratiometric and homogeneous assay for KANA detection based on microchip electrophoresis (MCE) was developed. First, by one single strand DNA (S-DNA) and one hairpin DNA (H-DNA), a novel R shape DNA probe (R-DNA) was prepared. After the probe was incubated with KANA, the S-DNA-KANA complex was formed, and H-DNA was released. Moreover, in the presence of exonuclease I (Exo-I), S-DNA-KANA complex would be digested to release the captured KANA for triggering target recycling and signal amplification. With the reaction going on, the fluorescence intensity of H-DNA (IH) increased and that of R-DNA (IR) decreased. They can be separated at different voltage intensities and converted to fluorescent signals for signal readout by MCE. The signal ratio of IH/IR was found to be linear toward target from 0.5 pg mL−1 to 10 ng mL−1, and the limit of detection was 150 fg mL−1. Moreover, it was successfully employed for KANA detection in milk and fish samples with consistent results of enzyme linked immune sorbent assay (ELISA). The R-DNA probe can quantitatively convert the amount of target to the intensity of DNA without label by MCE, and achieved exonuclease assisted signal amplification in homogenous solution. It was valuable to detect antibiotics residues in foodstuff with complex matrix. This approach broadened the application field of MCE to detect antibiotics without derivatization, which provided a promising platform for rapid screening of antibiotic residues in food.

Gold nanoparticles coupled with graphene quantum dots in organized medium to quantify aminoglycoside anti-biotics in yellow fever vaccine after solid phase extraction using a selective imprinted polymer

The determination of kanamycin sulfate was made indirectly by measuring its effect on photoluminescent amino functionalized graphene quantum dots (GQDs-amino) associated with gold nanoparticles (AuNPs) that were produced by the reduction of AuCl4 with NaBH4 in an aqueous dispersion of GQDs-amino (obtained by the pyrolysis of citric acid and glutathione) also containing the cationic surfactant CTAB. The AuNPs-GQDs-amino-CTAB system presents a suppressed photoluminescence that is amplified in the presence of kanamycin. Under optimized experimental conditions, the photoluminescence amplification of the nanomaterial system showed a linear response as a function of kanamycin concentration, covering three orders of magnitude (10−7 to 10−5 mol L−1). The use of solid phase extraction with a cartridge packed with aminoglycoside selective molecularly imprinted polymer ensured selectivity in determinations made on yellow-fever vaccine and veterinary pharmaceutical formulations. The analytical results were statistically similar to those obtained with an HPLC-based fluorescence method (after chemical derivatization). The proposed method is a simple, sensitive and selective approach that does not involve the use of toxic reagents employed for chemical derivatization of aminoglycoside antibiotics.

A dual-signal amplification strategy for kanamycin based on ordered mesoporous carbon-chitosan/gold nanoparticles-streptavidin and ferrocene labelled DNA

An ultrasensitive electrochemical aptasensor for kanamycin (KAN) detection was constructed with a dual-signal amplification strategy. The aptasensor achieved greatly amplified sensitivity due to the excellent electrical conductivity of the ordered mesoporous carbon-chitosan (OMC-CS)/gold nanoparticles-streptavidin (AuNPs-SA) and DNA2 labelled with ferrocene (Fc-DNA2). The AuNPs-SA was used to immobilize the DNA strand (biotin labelled) with the biotin-streptavidin system. The DNA2 strand containing the KAN aptamer was labelled with ferrocene to increase the current signal on the electrode surface when bound to KAN. Some factors that affect the performance of the aptasensor were optimized, and the proposed aptasensor provided a wide linear range from 1 × 10−10 M to 4 × 10−6 M, with a detection limit as low as 35.69 pM for KAN under the optimized conditions. This aptasensor had satisfactory electrochemical performance with good stability, sensitivity and reproducibility. Additionally, it also displayed a good specificity for KAN without interference from competitive analogues. Furthermore, the constructed aptasensor was successfully used to detect KAN in a real milk sample. The proposed method for KAN detection has great potential for the detection of other antibiotics.

Acclimatization of Pisum sativum L., grown in soil contaminated with veterinary antibiotics, an attribute of dose hormetic response of root metabolites

Plant-veterinary antibiotic interaction has been widely studied, however, to the best of our knowledge acclimatization studies with regard to changes in plant root metabolites has not been reported so far. The purpose of this study was to examine the changes in the metabolome of pea roots under antibiotic stress and their role in acclimatization. Pisum sativum L. was grown in soil contaminated with three commonly used veterinary antibiotics — kanamycin (KA), sulfamethazine (SA), and tetracycline (TC). In response to antibiotic stress, plants accumulated different types of low molecular weight compounds that provided protection from stress by contributing to ROS detoxification, protection of membrane integrity, efficient signaling, cell wall function, and cellular osmotic adjustment (glucose, galactose, myo-inositol, stigmasterol, octadecadienoic acid, l-proline). The concentration of amino acid, sugar, and triglyceride metabolites in KA and TC samples showed a dose-dependent biphasic (hormesis) fluctuation. This was mirrored in the metabolite abundance as well as the physiological attributes (mycorrhizal colonization, GST function, nutrient assimilation), which helped in the acclimatization without the loss of normal plant function. SA, on the other hand, had progressive toxic effects with increasing concentration. PCA revealed the differences to be due to SA treatments and in sterol and terpenoid metabolites.

Multiplexed aptasensor based on metal ions labels for simultaneous detection of multiple antibiotic residues in milk

A dual-target electrochemical aptasensor was developed for the simultaneous detection of multiple antibiotics based on metal ions as signal tracers and nanocomposites as signal amplification strategy. Metal ions such as Cd2+ and Pb2+ could generate distinct differential pulse voltammetry (DPV) peaks. When targets were present, kanamycin (KAN) and streptomycin (STR) as models, the KAN aptamer (KAP) and STR aptamer (STP) were released from their complementary strands, with more change of Cd2+ and Pb2+ corresponding to peak currents. At the same time, complementary strand of KAP (cKAP) and STP (cSTP) were linked with the poly (A) structure (cSTP-PolyA-cKAP) to increase their conformational freedom. Graphitized multi-walled carbon nanotubes (MWCNTGr) and carbon nanofibers-gold nanoparticles (CNFs-AuNPs) as a biosensor platform enhanced the surface area to capture a large amount of cSTP-PolyA-cKAP, thus amplifying the detection response. Under the optimal conditions, the aptasensor could detect KAN and STR as low as 74.50 pM and 36.45 pM respectively with the range from 0.1 to 100 nM and exhibited excellent selectively. Moreover, this aptasensor showed promising applications for the detection of other analytes by changing corresponding aptamers.

Diagnostic accuracy of GenoType® MTBDRsl VER 2.0 in detecting second-line drug resistance to M. tuberculosis.

A tertiary care hospital in North India. To evaluate the GenoType® MTBDRsl VER 2.0 assay for rapid diagnosis of second-line drug resistance to Mycobacterium tuberculosis. The MTBDRsl VER 2.0 assay was performed on 431 multidrug-resistant M. tuberculosis clinical isolates and specimens. The results were compared with phenotypic drug susceptibility testing (DST) and DNA sequencing. Molecular characterisation of drug resistance using DNA sequencing was performed for gyrA, gyrB, rrs and eis. Of the 415 isolates, respectively 176 (42.4%) and 40 (9.6%) were resistant to levofloxacin (LVX) and kanamycin (KM). The sensitivity and specificity of MTBDRsl VER 2.0 compared with phenotypic DST in detecting LVX resistance were respectively 97.2% (95%CI 93.5-99.1) and 99.1% (95%CI 97-99.9), and for KM resistance they were respectively 92.5% (95%CI 79.6-98.4) and 99.5% (95%CI 98.1-99.9). The MTBDRsl VER 2.0 assay showed very high sensitivity and specificity for the detection of second-line drug resistance, suggesting it has potential for the rapid, early detection of such cases.

Potent 1,2,4-Triazino[5,6 b]indole-3-thioether Inhibitors of the Kanamycin Resistance Enzyme Eis from Mycobacterium tuberculosis.

A common cause of resistance to kanamycin (KAN) in tuberculosis is overexpression of the enhanced intracellular survival (Eis) protein. Eis is an acetyltransferase that multiacetylates KAN and other aminoglycosides, rendering them unable to bind the bacterial ribosome. By high-throughput screening, a series of substituted 1,2,4-triazino[5,6 b]indole-3-thioether molecules were identified as effective Eis inhibitors. Herein, we purchased 17 and synthesized 22 new compounds, evaluated their potency, and characterized their steady-state kinetics. Four inhibitors were found not only to inhibit Eis in vitro, but also to act as adjuvants of KAN and partially restore KAN sensitivity in a Mycobacterium tuberculosis KAN-resistant strain in which Eis is upregulated. A crystal structure of Eis in complex with a potent inhibitor and CoA shows that the inhibitors bind in the aminoglycoside binding site snugly inserted into a hydrophobic cavity. These inhibitors will undergo preclinical development as novel KAN adjuvant therapies to treat KAN-resistant tuberculosis.