169,781 research outputs found
Using detergent to enhance detection sensitivity of African trypanosomes in human CSF and blood by Loop-Mediated Isothermal Amplification (LAMP)
No full text<p><b>Background:</b> The loop-mediated isothermal amplification (LAMP) assay, with its advantages of simplicity, rapidity and cost effectiveness, has evolved as one of the most sensitive and specific methods for the detection of a broad range of pathogenic microorganisms including African trypanosomes. While many LAMP-based assays are sufficiently sensitive to detect DNA well below the amount present in a single parasite, the detection limit of the assay is restricted by the number of parasites present in the volume of sample assayed; i.e. 1 per µL or 103 per mL. We hypothesized that clinical sensitivities that mimic analytical limits based on parasite DNA could be approached or even obtained by simply adding detergent to the samples prior to LAMP assay.</p>
<p><b>Methodology/Principal Findings:</b> For proof of principle we used two different LAMP assays capable of detecting 0.1 fg genomic DNA (0.001 parasite). The assay was tested on dilution series of intact bloodstream form Trypanosoma brucei rhodesiense in human cerebrospinal fluid (CSF) or blood with or without the addition of the detergent Triton X-100 and 60 min incubation at ambient temperature. With human CSF and in the absence of detergent, the LAMP detection limit for live intact parasites using 1 µL of CSF as the source of template was at best 103 parasites/mL. Remarkably, detergent enhanced LAMP assay reaches sensitivity about 100 to 1000-fold lower; i.e. 10 to 1 parasite/mL. Similar detergent-mediated increases in LAMP assay analytical sensitivity were also found using DNA extracted from filter paper cards containing blood pretreated with detergent before card spotting or blood samples spotted on detergent pretreated cards.</p>
<p><b>Conclusions/Significance:</b> This simple procedure for the enhanced detection of live African trypanosomes in biological fluids by LAMP paves the way for the adaptation of LAMP for the economical and sensitive diagnosis of other protozoan parasites and microorganisms that cause diseases that plague the developing world.</p>
Characterization of Acoustic Resonance in a High Pressure Sodium Lamp
No full textWith the last decades, the high pressure sodium (HPS) lamp has been supplied in high frequency in order to increase the efficacy of the lamp/ballast system. However, at some given frequencies, standing acoustic waves, namely acoustic resonance (AR), might develop in the burner and cause lamp luminous fluctuation, extinction and destruction in the most serious case. As we seek for a control method to detect and avoid the lamp AR some main characteristics of the acoustic resonances in a 150W HPS lamp are presented in this paper,. The first one is the characteristic of the lamp AR threshold power, the second one is the differences between forward and backward frequency scanning effects during lamp open loop operation. Thirdly, lamp AR behaviour in closed loop operation with an LCC half bridge inverter will be presented and leads to a new point of view and a change in the choice of the AR detection method. These characteristics allow us to further understand the AR and to better control the lamp
Loop-mediated isothermal amplification (LAMP) for point-of-care detection of asymptomatic low-density malaria parasite carriers in Zanzibar.
No full textBACKGROUND: Asymptomatic, low parasite density malaria infections are difficult to detect with currently available point-of-care diagnostics. This study piloted a loop-mediated isothermal amplification (LAMP) kit for field-friendly, high-throughput detection of asymptomatic malaria infections during mass screening and treatment (MSAT) in Zanzibar, a malaria pre-elimination setting. METHODS: Screening took place in three known hotspot areas prior to the short rains in November. Finger-prick blood was taken for screening by rapid diagnostic test (RDT) and LAMP and collected on filter paper for subsequent polymerase chain reaction (PCR) analyses. LAMP results were compared to RDT and to PCR using McNemar's test. RESULTS: Approximately 1,000 people were screened. RDT detected ten infections (1.0% (95% CI 0.3-1.6)) whilst both LAMP and PCR detected 18 (1.8% (95% CI 0.9-2.6)) infections. However, PCR identified three infections that LAMP did not detect and vice versa. LAMP testing was easy to scale-up in field conditions requiring minimal training and equipment, with results ready one to three hours after screening. CONCLUSIONS: Despite lower than expected prevalence, LAMP detected a higher number of infections than the currently used diagnostic, RDT. LAMP is a field-friendly, sensitive diagnostic test that could be useful for MSAT malaria campaigns which require quick results to enable prompt treatment
Diagnostic accuracy of loop-mediated isothermal amplification (LAMP) for detection of Leishmania DNA in buffy coat from visceral leishmaniasis patients
No full textBACKGROUND: Visceral leishmaniasis (VL) remains as one of the most neglected tropical diseases with over 60% of the world's total VL cases occurring in the Indian subcontinent. Due to the invasive risky procedure and technical expertise required in the classical parasitological diagnosis, the goal of the VL experts has been to develop noninvasive procedure(s) applicable in the field settings. Several serological and molecular biological approaches have been developed over the last decades, but only a few are applicable in field settings that can be performed with relative ease. Recently, loop-mediated isothermal amplification (LAMP) has emerged as a novel nucleic acid amplification method for diagnosis of VL. In this study, we have evaluated the LAMP assay using buffy coat DNA samples from VL patients in Bangladesh and compared its performance with leishmania nested PCR (Ln-PCR), an established molecular method with very high diagnostic indices. METHODS: Seventy five (75) parasitologically confirmed VL patients by spleen smear microcopy and 101 controls (endemic healthy controls -25, healthy control-26, Tuberculosis-25 and other diseases-25) were enrolled in this study. LAMP assay was carried out using a set of four primers targeting L. donovani kinetoplast minicircle DNA under isothermal (62 [degree sign]C) conditions in a heat block. For Ln-PCR, we used primers targeting the parasite's small-subunit rRNA region. RESULTS: LAMP assay was found to be positive in 68 of 75 confirmed VL cases, and revealed its diagnostic sensitivity of 90.7% (95.84-81.14, 95% CI), whereas all controls were negative by LAMP assay, indicating a specificity of 100% (100--95.43, 95% CI). The Ln-PCR yielded a sensitivity of 96% (98.96-87.97, 95% CI) and a specificity of 100% (100--95.43, 95% CI). CONCLUSION: High diagnostic sensitivity and excellent specificity were observed in this first report of LAMP diagnostic evaluation from Bangladesh. Considering its many fold advanta over conventional PCR and potential to be used as a simple and rapid test in the VL endemic areas of the Indian subcontinent, our findings are encouraging, but further evaluation of LAMP is neede
Loop-mediated isothermal amplification (LAMP) test for detection of Trypanosoma evansi strain B
No full textCamel Trypanosomiasis (Surra) is mainly caused by Trypanosoma evansi strains that express variable surface glycoprotein (VSG) RoTat 1.2 However, in Kenya a second causative strain that does not express RoTat 1.2 VSG (T. evansi type B) has been identified. The prevalence of T. evansi type B largely remains unknown due to inadequate diagnostic assay. This work reports the development of a sensitive and specific diagnostic assay capable of detecting T. evansi type B based on the strategy of Loop-mediated Isothermal Amplification (LAMP) of DNA. The test is rapid and amplification is achieved within 20-25min at 63°C using a real time PCR machine. Restriction enzyme AluI digestion of the amplicon gave the predicted 83bp and 89bp sized bands and the LAMP product melt curves showed consistent melting temperature (Tm) of ∼89°C. The assay analytical sensitivity is ∼0.1tryps/ml while that of classical PCR test targeting the same gene is ∼10tryps/ml. There was a 100% agreement in detection of the LAMP amplification product in real time, gel electrophoresis, on addition of SYBR Green I, and when using chromatographic Lateral Flow Dipstick (LFD) format. The use of the LAMP test revealed nine more T. evansi type B DNA samples that were not initially detected through PCR. The robustness and higher sensitivity of the T. evansi type B LAMP assay coupled with the visual detection of the amplification product indicate that the technique has strong potential as a point-of-use test in surra endemic areas
Loop-mediated isothermal amplification (LAMP) method for rapid detection of Trypanosoma brucei rhodesiense
No full textLoop-mediated isothermal amplification (LAMP) of DNA is a novel technique that rapidly amplifies target DNA under isothermal conditions. In the present study, a LAMP test was designed from the serum resistance-associated (SRA) gene of Trypanosoma brucei rhodesiense, the cause of the acute form of African sleeping sickness, and used to detect parasite DNA from processed and heat-treated infected blood samples. The SRA gene is specific to T. b. rhodesiense and has been shown to confer resistance to lysis by normal human serum. The assay was performed at 62°C for 1 h, using six primers that recognised eight targets. The template was varying concentrations of trypanosome DNA and supernatant from heat-treated infected blood samples. The resulting amplicons were detected using SYTO-9 fluorescence dye in a real-time thermocycler, visual observation after the addition of SYBR Green I, and gel electrophoresis. DNA amplification was detected within 35 min. The SRA LAMP test had an unequivocal detection limit of one pg of purified DNA (equivalent to 10 trypanosomes/ml) and 0.1 pg (1 trypanosome/ml) using heat-treated buffy coat, while the detection limit for conventional SRA PCR was ∼1,000 trypanosomes/ml. The expected LAMP amplicon was confirmed through restriction enzyme RsaI digestion, identical melt curves, and sequence analysis. The reproducibility of the SRA LAMP assay using water bath and heat-processed template, and the ease in results readout show great potential for the diagnosis of T. b. rhodesiense in endemic regions
[Report to Chief J. E. Curry, by an unknown author #1]
No full textReport to Chief J. E. Curry, by an unknown author. The report contains a list of officers who gave depositions to the United States Attorney
[Report to Chief J. E. Curry, by an unknown author #2]
No full textReport to Chief J. E. Curry, by an unknown author. The report contains a list of officers who gave depositions to the United States Attorney
Sources d'Alimentation Électrique pour l'Étude et l'Utilisation Efficace des Lampes Excimer DBD
No full textAvec l'objectif d'améliorer le rendement des lampes à excimères (Excilampe) à décharge à barrière diélectrique (DBD), un convertisseur en mode de courant, qui permet un ajustement précis de la puissance électrique injectée dans ce type des lampes, à été conçu et mis en oeuvre. Ce convertisseur fournit à la lampe un courant de forme d'onde carrée contrôlé au moyen de trois paramètres: l'amplitude, la fréquence et le rapport cyclique, pour obtenir un contrôle total de l'énergie électrique transmise à l'excilampe DBD. La mise en oeuvre intègre un transformateur élévateur comme interface entre la lampe et un commutateur. Les expériences démontrent le principe de fonctionnement de ce convertisseur, y compris les mesures de puissance du rayonnement UV. Les degrés de liberté du convertisseur sont utilisées pour analyser le comportement de la lampe sous différentes combinaisons de ces trois paramètres, et sont utilisés pour déterminer le point de fonctionnement optimal de la lampe. Ensuite, un convertisseur à résonance du type onduleur série, est proposé pour alimenter la lampe avec une grande efficacité électrique. Afin de contrôler effectivement la puissance de la lampe, le mode de fonctionnement de ce convertisseur utilise le mode de conduction discontinue et la commutation douce (ZCS), avec lequel on obtient aussi de faibles émissions électromagnétiques et l'on réduit les pertes de commutation. Les relations mathématiques obtenus à partir de l'analyse du diagramme de phase, ont été validées par des simulations et avec des résultats expérimentaux. Enfin, différentes topologies d'alimentations pour DBD sont comparées analytiquement et expérimentalement pour évaluer objectivement les avantages de chaque approche. Une des perspectives de ce travail est l'application de l'alimentation en créneaux pour l'étude de la performance d'autres types de réacteurs et d'excilampes DBD. ABSTRACT : With the aim to provide a scientific tool for the enhancement of the Dielectric Barrier Discharge (DBD) Excimer Lamps (Excilamp) performance, a current-mode converter that allows an accurate adjustment of the electrical power injected into one of those lamps, is designed and implemented. With the proposed converter, the current supplied to the lamp has a square shape, controlled by means of three parameters: amplitude, duty cycle and frequency, which provides full control of the lamp electrical power. Implementation is made considering a step-up transformer interfacing the high-voltage lamp with the converter. Experiments demonstrate the operating principle of this converter, including UV power measurements for a DBD XeCl Excilamp. The capabilities of the converter are used to analyze the lamp behavior under different combinations of these three parameters, illustrating its capabilities for finding the optimal operating point. Then a series-resonant inverter for the supply of DBD) excilamp is proposed. In order to effectively control the lamp power, the operating mode of this converter combines discontinuous current-mode and soft-commutation (ZCS), obtaining as well low electromagnetic emissions, and reduced switching losses. The mathematical relationships obtained from state plane analysis, are validated with simulations and experimental results. Finally, several topologies of DBDs power supplies are compared analytical and experimentally to elucidate the advantages of each approach. After this work, one of the perspectives is the application of the square-shape supply in the performance study of other types of DBD excilamps and DBD reactors
Arc Lamp Attachment
No full textPatent for improving arc lamp attachments. The attachment allows an arc lamp to be able to use an incandescent lamp in place of the electrodes in an arc lamp. The attachment increases the efficiency and durability of the arc lamp
- …
