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1.
Diagn Microbiol Infect Dis ; 92(4): 299-304, 2018 Dec.
Article in English | MEDLINE | ID: mdl-30025968

ABSTRACT

BACKGROUND: Top priorities for tuberculosis control and elimination include a simple, low-cost screening test using sputum and a non-sputum-based test in patients that do not produce sputum. The aim of this study was to evaluate the performance of a colorimetric sensor array (CSA) test, for analysis of volatile organic compounds in urine, in the diagnosis of pulmonary TB. MATERIAL AND METHODS: Urine samples were collected from individuals suspected of having pulmonary TB in Western Kenya. Reference methods included MGIT culture and/or Xpert MTB/RIF nucleic acid amplification test on sputa. Fresh urine samples were tested with the CSA, with acid and base and without an additive. The CSA were digitally imaged, and the resulting colorimetric response patterns were used for chemometric analysis. Sensitivity, specificity, and negative (NPV) and positive predictive (PPV) values were determined for HIV-positive and HIV-negative patients. RESULTS: In HIV-negative patients, the highest accuracy was obtained in urine samples pre-treated with a base, yielding a sensitivity, specificity, PPV, and NPV of 78.3% (65/83), 69.2% (54/78), 73.0% (n/89) and 75.0% (n/72). The highest sensitivity of 79.5% was achieved using sensor data from all three test conditions at a specificity of 65.4%. In HIV-positive subjects, the sensor performance was substantially lower with sensitivity, specificity, PPV, and NPV ranging from 48.3% to 62.3%, 54.1% to 74.0%, 55.9% to 64.2%, and 60.6% to 64.9%, respectively. CONCLUSION: The CSA fingerprint of urine headspace volatiles showed moderate accuracy in diagnosing TB in HIV-negative patients, but the sensor performance dropped substantially in HIV-coinfected patients. Further development of TB-responsive CSA indicators may improve the accuracy of CSA urine assay.


Subject(s)
Colorimetry/methods , Mycobacterium tuberculosis , Tuberculosis/diagnosis , Tuberculosis/urine , Volatile Organic Compounds/urine , Case-Control Studies , Coinfection , Female , HIV Infections , Humans , Interferon-gamma Release Tests , Male , Mycobacterium tuberculosis/isolation & purification , Mycobacterium tuberculosis/metabolism , Reproducibility of Results , Sensitivity and Specificity , Sputum/microbiology , Tuberculosis/microbiology , Tuberculosis, Pulmonary/diagnosis , Tuberculosis, Pulmonary/urine
2.
Analyst ; 141(3): 918-25, 2016 Feb 07.
Article in English | MEDLINE | ID: mdl-26753182

ABSTRACT

Clinical microbiology automation is currently limited by the lack of an in-plate culture identification system. Using an inexpensive, printed, disposable colorimetric sensor array (CSA) responsive to the volatiles emitted into plate headspace by microorganisms during growth, we report here that not only the presence but the species of bacteria growing in plate was identified before colonies are visible. In 1894 trials, 15 pathogenic bacterial species cultured on blood agar were identified with 91.0% sensitivity and 99.4% specificity within 3 hours of detection. The results indicate CSAs integrated into Petri dish lids present a novel paradigm to speciate microorganisms, well-suited to integration into automated plate handling systems.


Subject(s)
Electronic Nose , Gram-Negative Bacteria/isolation & purification , Gram-Positive Bacteria/isolation & purification , Molecular Probe Techniques/instrumentation , Volatile Organic Compounds/analysis , Reproducibility of Results
3.
J Breath Res ; 8(2): 027112, 2014 Jun.
Article in English | MEDLINE | ID: mdl-24862241

ABSTRACT

Cancer diagnosis is typically delayed to the late stages of disease due to the asymptomatic nature of cancer in its early stages. Cancer screening offers the promise of early cancer detection, but most conventional diagnostic methods are invasive and remain ineffective at early detection. Breath analysis is, however, non-invasive and has the potential to detect cancer at an earlier stage by analyzing volatile biomarkers in exhaled breath. This paper summarizes breath sampling techniques and recent developments of various array-based sensor technologies for breath analysis. Significant advancements were made by a number of different research groups in the development of nanomaterial-based sensor arrays, and the ability to accurately distinguish cancer patients from healthy controls based on the volatile organic compounds (VOCs) in exhaled breath has been demonstrated. Optical sensors based on colorimetric sensor array technology are also discussed, where preliminary clinical studies suggest that metabolic VOC profiles could be used to accurately diagnose various forms of lung cancer. Recent studies have demonstrated the potential of using metabolic VOCs for cancer detection, but further standardization and validation is needed before breath analysis can be widely adopted as a clinically useful tool.


Subject(s)
Breath Tests/instrumentation , Breath Tests/methods , Chemistry Techniques, Analytical/instrumentation , Chemistry Techniques, Analytical/methods , Neoplasms/diagnosis , Volatile Organic Compounds/analysis , Colorimetry , Data Interpretation, Statistical , Humans , Neoplasms/metabolism
4.
J Clin Microbiol ; 52(2): 592-8, 2014 Feb.
Article in English | MEDLINE | ID: mdl-24478493

ABSTRACT

Sepsis is a medical emergency demanding early diagnosis and tailored antimicrobial therapy. Every hour of delay in initiating effective therapy measurably increases patient mortality. Blood culture is currently the reference standard for detecting bloodstream infection, a multistep process which may take one to several days. Here, we report a novel paradigm for earlier detection and the simultaneous identification of pathogens in spiked blood cultures by means of a metabolomic "fingerprint" of the volatile mixture outgassed by the organisms. The colorimetric sensor array provided significantly faster detection of positive blood cultures than a conventional blood culture system (12.1 h versus 14.9 h, P < 0.001) while allowing for the identification of 18 bacterial species with 91.9% overall accuracy within 2 h of growth detection. The colorimetric sensor array also allowed for discrimination between unrelated strains of methicillin-resistant Staphylococcus aureus, indicating that the metabolomic fingerprint has the potential to track nosocomial transmissions. Altogether, the colorimetric sensor array is a promising tool that offers a new paradigm for diagnosing bloodstream infections.


Subject(s)
Bacteria/isolation & purification , Bacteriological Techniques/methods , Biosensing Techniques/methods , Blood Chemical Analysis/methods , Blood/microbiology , Colorimetry/methods , Sepsis/diagnosis , Bacteria/classification , Humans , Metabolomics/methods , Sepsis/microbiology , Time
5.
PLoS One ; 8(5): e62726, 2013.
Article in English | MEDLINE | ID: mdl-23671629

ABSTRACT

A colorimetric sensor array is a high-dimensional chemical sensor that is cheap, compact, disposable, robust, and easy to operate, making it a good candidate technology to detect pathogenic bacteria, especially potential bioterrorism agents like Yersinia pestis and Bacillus anthracis which feature on the Center for Disease Control and Prevention's list of potential biothreats. Here, a colorimetric sensor array was used to continuously monitor the volatile metabolites released by bacteria in solid media culture in an Advisory Committee on Dangerous Pathogen Containment Level 3 laboratory. At inoculum concentrations as low as 8 colony-forming units per plate, 4 different bacterial species were identified with 100% accuracy using logistic regression to classify the kinetic profile of sensor responses to culture headspace gas. The sensor array was able to further discriminate between different strains of the same species, including 5 strains of Yersinia pestis and Bacillus anthracis. These preliminary results suggest that disposable colorimetric sensor arrays can be an effective, low-cost tool to identify pathogenic bacteria.


Subject(s)
Bacteria/metabolism , Biosensing Techniques/methods , Colorimetry/methods , Gases/analysis , Bacillus anthracis/growth & development , Bacillus anthracis/metabolism , Bacteria/classification , Bacteria/growth & development , Bacterial Typing Techniques/methods , Bioterrorism/prevention & control , Culture Media/metabolism , Gases/chemistry , Gases/metabolism , Logistic Models , Reproducibility of Results , Species Specificity , Yersinia pestis/growth & development , Yersinia pestis/metabolism
6.
Phys Chem Chem Phys ; 14(31): 11027-39, 2012 Aug 21.
Article in English | MEDLINE | ID: mdl-22766969

ABSTRACT

The development of soft bioelectronic interfaces with accurate compositional and topological control of the supramolecular architecture attracts intense interest in the fast-growing field of bioelectronics and biosensing. The present study explores the recognition-driven layer-by-layer assembly of glycoenzymes onto electrode surfaces. The design of the multi-protein interfacial architecture is based on the multivalent supramolecular carbohydrate-lectin interactions between redox glycoproteins and concanavalin A (Con A) derivatives. Specifically, [Os(bpy)(2)Clpy](2+)-tagged Con A (Os-Con A) and native Con A were used to direct the assembly of horseradish peroxidase (HRP) and glucose oxidase (GOx) in a stepwise topologically controlled procedure. In our designed configuration, GOx acts as the biorecognition element to glucose stimulus, while HRP acts as the transducing element. Surface plasmon resonance (SPR) spectroscopy and quartz crystal microbalance with dissipation (QCM-D) results are combined to give a close representation of the protein surface coverage and the content of water in the protein assembly. The characterization is complemented with in situ atomic force microscopy (AFM) to give a topographical description of the layers assemblage. Electrochemical (EC) techniques were used to characterize the functional features of the spontaneously self-assembled biohybrid architecture, showing that the whole system presents efficient electron transfer and mass transport processes being able to transform micromolar glucose concentration into electrical information. In this way the combination of the electroactive and nonelectroactive Con A provides an efficient strategy to control the position and composition of the protein layers via recognition-driven processes, which defines its sensitivity toward glucose. Furthermore, the incorporation of dextran as a permeable interlayer able to bind Con A promotes the physical separation of the biochemical and transducing processes, thus enhancing the magnitude of the bioelectrochemical signal. We consider that these results are relevant for the nanoconstruction of functional biointerfaces provided that our experimental evidence reveals the possibility of locally addressing recognition, transduction and amplification elements in interfacial ensembles via LbL recognition-driven processes.


Subject(s)
Concanavalin A/metabolism , Glucose Oxidase/metabolism , Horseradish Peroxidase/metabolism , Biosensing Techniques , Concanavalin A/chemistry , Dextrans/chemistry , Electrochemical Techniques , Electrodes , Glucose/chemistry , Glucose/metabolism , Glucose Oxidase/chemistry , Gold/chemistry , Horseradish Peroxidase/chemistry , Oxidation-Reduction , Protein Binding , Quartz Crystal Microbalance Techniques , Surface Plasmon Resonance , Surface Properties , Water/chemistry
8.
Langmuir ; 26(16): 13684-96, 2010 Aug 17.
Article in English | MEDLINE | ID: mdl-20695621

ABSTRACT

The convergence of chemistry, biology, and materials science has paved the way to the emergence of hybrid nanobuilding blocks that incorporate the highly selective recognition properties of biomolecules, with the tailorable functional capabilities of inorganic molecules. In this work, we describe for the first time the decoration of concanavalin A (Con A), a protein with the ability to recognize sugars and form glycoconjugates, with Os(II) redox-active complexes. This strategy enabled the construction of electroactive biosupramolecular materials whose redox potentials could be easily modulated through the facile molecular modification of the electroactive inorganic complexes. Small-angle X-ray scattering (SAXS), steady-state fluorescence, surface plasmon resonance (SPR) spectroscopy, matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-TOF-MS), and differential-pulsed (DPV) and cyclic voltammetry (CV) were used to characterize the structural and functional features of the synthesized biohybrid building blocks as well as their respective supramolecular assemblies built up on gold electrodes. By harnessing the electroactive and carbohydrate-recognition properties of these tailor-made biohybrid building blocks, we were able to integrate glucose oxidase (GOx) onto gold electrodes via sugar-lectin interactions. The redox activity of the Os-modified Con A interlayer allowed the electronic connection between the multilayered GOx assemblies and the metal electrode as evidenced by the well-defined bioelectrocatalytic response exhibited by the biomolecular assemblies in the presence of the glucose in solution. We consider that this approach based on the spontaneous formation of redox-active biosupramolecular assemblies driven by recognition processes can be of practical relevance for the facile design of biosensors, as well as for the construction of new multifunctional bioelectrochemical systems.


Subject(s)
Concanavalin A/chemistry , Nanostructures/chemistry , Electrochemistry , Models, Theoretical , Scattering, Small Angle , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization , Surface Plasmon Resonance
9.
Phys Chem Chem Phys ; 12(28): 8071-83, 2010 Jul 28.
Article in English | MEDLINE | ID: mdl-20526515

ABSTRACT

There is a growing quest for the construction of functional supramolecular architectures to efficiently translate (bio)chemical events into easily measurable signals. This interest originates from its inherent scientific relevance as well as from their potential applications in the ever-flourishing areas of bioelectronics and biosensing. Herein, we describe the immobilization of glycoproteins onto electrode surfaces based on recognition-mediated supramolecular processes. Quartz crystal microbalance with dissipation (QCM-D), surface plasmon resonance (SPR) spectroscopy, and electrochemical (EC) measurements were used to characterize the structural and functional features of these bio-supramolecular systems. Carbohydrate-lectin interactions were successfully used to build up stable assemblies of glucose oxidase (GOx) layers mediated by the recognition properties of concanavalin A supramolecular architectures. The catalytic response of GOx indicates that the whole population of enzymes incorporated in the supramolecular architecture is fully active. Even though lectin-carbohydrate interactions are rather weak, the multivalency effects prevailing in the supramolecular assembly confer remarkable stability to the interfacial architecture, thus preventing the release of the enzyme from the surface even with high glucose (ligand) concentrations. This approach represents a simple and straightforward route to locally address functional glycoproteins at interfaces. In this context, we consider that the versatility of a supramolecular assembly using biological interactions could open up new ways of envisioning or to generate new ideas for the future development of highly efficient bioelectronic platforms.


Subject(s)
Concanavalin A/chemistry , Glucose Oxidase/chemistry , Gold/chemistry , Electrochemical Techniques , Electrodes , Glucose Oxidase/metabolism , Kinetics , Surface Plasmon Resonance
10.
Proc Natl Acad Sci U S A ; 105(34): 12134-9, 2008 Aug 26.
Article in English | MEDLINE | ID: mdl-18711145

ABSTRACT

The development of low-cost, reliable sensors will rely on devices capable of converting an analyte binding event to an easily read electrical signal. Organic thin-film transistors (OTFTs) are ideal for inexpensive, single-use chemical or biological sensors because of their compatibility with flexible, large-area substrates, simple processing, and highly tunable active layer materials. We have fabricated low-operating voltage OTFTs with a cross-linked polymer gate dielectric, which display stable operation under aqueous conditions over >10(4) electrical cycles using the p-channel semiconductor 5,5'-bis-(7-dodecyl-9H-fluoren-2-yl)-2,2'-bithiophene (DDFTTF). OTFT sensors were demonstrated in aqueous solutions with concentrations as low as parts per billion for trinitrobenzene, methylphosphonic acid, cysteine, and glucose. This work demonstrates of reliable OTFT operation in aqueous media, hence opening new possibilities of chemical and biological sensing with OTFTs.


Subject(s)
Biosensing Techniques/methods , Transistors, Electronic/standards , Cysteine/analysis , Glucose/analysis , Organic Chemicals/analysis , Organophosphorus Compounds/analysis , Trinitrobenzenes/analysis , Water
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