Novel Method For Rapid Detection Research Articles

A novel method for rapid detection of a Helicobacter pylori infection using a \u03b3-glutamyltranspeptidase-activatable fluorescent probe

A γ-glutamyl hydroxymethyl rhodamine green probe (gGlu-HMRG) reacts with γ-glutamyltranspeptidase (GGT) and immediately produces fluorescence, is clinically applied for real-time cancers’ visualization. Since Helicobacter pylori produces GGT, this study aimed to investigate whether gGlu-HMRG can be used to detect H. pylori infections. A wild-type H. pylori strain and the ggt gene-disrupted mutant were cultured and treated with gGlu-HMRG. This fluorescent probe assay was used to quantify GGT activity of H. pylori ex vivo using gastric biopsy specimens. The H. pylori diagnostic capabilities of the assay were determined from altered fluorescence intensity (FI) values at 5 min (FIV-5) and 15 minutes (FIV-15). Distinct fluorescence was identified in wild H. pylori strain, using gGlu-HMRG, whereas no fluorescence was observed in ggt gene-disrupted mutant strain. On ex vivo imaging of gGlu-HMRG, fluorescence intensity increased markedly with time in H. pylori-positive specimens; however, the H. pylori-negative specimens displayed a slight increase in FI. FIV-5 and FIV-15 differed significantly between H. pylori-positive and -negative specimens. FIV-15 differed significantly between H. pylori-positive and -eradicated group. This assay sensitivity and specificity were 75.0% and 83.3% in the antrum and 82.6% and 89.5% in the stomach body. GGT-activatable fluorescence probe is applicable for rapid diagnosis of H. pylori.

Novel Method for Rapid Detection of Spatiotemporal HIV Clusters Potentially Warranting Intervention.

Rapid detection of increases in HIV transmission enables targeted outbreak response efforts to reduce the number of new infections. We analyzed US HIV surveillance data and identified spatiotemporal clusters of diagnoses. This systematic method can help target timely investigations and preventive interventions for maximum public health benefit.

Rapid detection of Escherichia coli in beverages using genetically engineered bacteriophage T7

Foodborne illness due to bacterial contamination is a significant issue impacting public health that demands new technology which is practical to implement by food industry. Detection of bacteria in food products and production facilities is a crucial strategy supporting food safety assessments. Bacteriophages were investigated as a tool for bacterial detection due to their ability to infect specific strain of host bacteria in order to improve sensitivity, specificity, and rapidity of bacterial detection. The results of this investigation reveal a novel method for rapid detection. The method employs a genetically engineered bacteriophage, phage T7-ALP, which expresses alkaline phosphatase. Upon infection of Escherichia coli, overexpression of alkaline phosphatase provides an opportunity for rapid sensitive detection of a signal indicative of bacterial presence in model beverage samples as low as 100 bacteria per gram. The method employs a fluorescent precipitated substrate, ELF-97, as a substrate for alkaline phosphatase activity coupled with fluorescence imaging and image analysis allowing single-cell imaging results in high detection sensitivity. The method is easily completed within less than 6 h enabling it to be deployed within most large industrial food processing facilities that have routine 8-h operational shifts.

Novel Method for Rapid Detection of Staphylococcus Aureus and Its Enterotoxins in Patients with Diarrhea

Novel Method for Rapid Detection of <i>Staphylococcus</i> <i>Aureus</i> and Its Enterotoxins in Patients with Diarrhea

Liquid array diagnostics: a novel method for rapid detection of microbial communities in single-tube multiplex reactions.

We present a novel liquid array diagnostics (LAD) method, which enables rapid and inexpensive detection of microbial markers in a single-tube multiplex reaction. We evaluated LAD both on pure cultures, and on infant gut microbiota for a 15-plex reaction. LAD showed more than 80% accuracy of classification and a detection limit lower than 2% of the Illumina reads per sample. The results on the clinical dataset showed that there was a rapid decrease of staphylococci from 10-day- to 4-month-old children, a peak of bifidobacteria at 4months, and a peak of Bacteroides in2-year-old children, which is in accordance with findings described in the literature. Being able to detect up to 50 biomarkers, LAD is a suitable method for assays where high throughput is essential.

Rapid and On-Site Detection of Uranyl Ions via Ratiometric Fluorescence Signals Based on a Smartphone Platform.

Fluorescent quantum dots (QDs) of carbon and semiconductors have superior optical properties and show great potential in sensing applications. This paper reports a novel method for rapid detection of uranyl ions via ratiometric fluorescence signals by employing two types of QDs as the key materials. As the most soluble and stable toxic uranium species, uranyl has been recognized as an important index for nuclear industrial wastewater. However, its on-site, rapid, and sensitive determination remains challenging. This work uses the ratiometric fluorescent signal of QDs and combines a smartphone-based handheld device for on-site and rapid detection of uranyl. The ratiometric fluorescent probe is achieved by integrating carbon dots (C-dots) and CdTe QDs (MPA@CdTe QDs) through chemical hybridization. The presence of uranyl ions greatly quenches the red fluorescence of the CdTe QDs, whereas the green fluorescence keeps constant, leading to an obvious color change. An app and a 3D-printed accessory have been developed on a smartphone to analyze and calculate the content of uranyl on the basis of captured fluorescence signals from a test strip with an immobilized probe. This new designed mobile detection system displays good analytical performance for uranyl ions in a wide concentration range of 1 to 150 μM, which shows a great potential application in controlling the nuclear industrial pollution.

Rapid Detection of Six Glucocorticoids Added Illegally to Dietary Supplements by Combining TLC with Spot-Concentrated Raman Scattering.

The objective of this study was to establish a novel method for rapid detection of six glucocorticoids (prednisone, prednisone acetate, prednisolone, hydrocortisone, hydrocortisone acetate, and dexamethasone) added illegally in dietary supplements simultaneously by combining thin layer chromatography (TLC) with spot-concentrated Raman scattering (SCRS). The doping ingredients were separated by TLC, and viewed and located with UV light (254 nm), enriched by chromatography, then Raman spectra were directly detected by a Raman Imagine microscope with 780 nm laser source. This method had complementary advantages of TLC and Raman spectroscopy, which enhanced the specificity of the test results. The limit of detection (LOD) of the reference substances were 4 μg, 4 μg, 4 μg, 6 μg, 6 μg, and 4 μg, respectively. The method was used to study the six glucocorticoids added illegally in five dietary supplements. Fake drugs had been detected. The study showed that the TLC-SCRS method is simple, rapid, specific, sensitive, and reliable. The method could be used for effective separation and detection of six chemical components used in dietary supplement products, and would have good prospects for on-site qualitative screening of dietary supplement products for adulterants.

A novel method for rapid detection of Streptococcus pneumoniae antigens in blood.

In this study, we used "RAPIRUN(®)Streptococcus pneumoniae HS (otitis media/sinusitis) (RAPIRUN-HS)," a rapid S. pneumoniae antigen detection kit, to investigate methods for detecting S. pneumoniae antigens in blood of 32 bacterial pneumonia patients. We simultaneously performed PCR to detect S. pneumoniae in blood samples. The results of these tests were compared based on pneumonia severity, determined using the Pneumonia Severity Index (PSI) score classification. Four S. pneumoniae PCR-positive patients of the six severe pneumococcal pneumonia patients (PSI risk class IV/V) also tested positive using RAPIRUN-HS. Twenty-four mild to moderate pneumonia patients (PSI risk class I-III) were S. pneumoniae PCR-negative; of these, 21 tested negative using RAPIRUN-HS. The pneumococcal pneumonia patients testing positive using RAPIRUN-HS had low leukocyte counts and elevated C-reactive protein and procalcitonin levels, indicating that RAPIRUN-HS results were correlated with pneumonia severity. The time course evaluations of the laboratory tests for severe pneumococcal pneumonia patients showed that RAPIRUN-HS and S. pneumoniae PCR yielded positive results earlier than the changes in procalcitonin and IL-6. Thus, concomitant pneumococcal bacteremia was strongly suspected in patients testing positive using RAPIRUN-HS. In conclusion, RAPIRUN-HS may be useful for determining whether to admit patients into hospitals and selecting the appropriate antimicrobial agents.

Lateral Flow Impedance Transducer for Salmonella Detection

In this work a novel method for rapid detection of Salmonella bacteria based upon impedance spectroscopy, and lateral flow membrane is introduced. To develop home based diagnostic devices, the systems need to be ease of use; low-cost; sensitive and rapid. In this work, arrays of interdigited gold micro-electrodes (IDE) were fabricated and targets specific antibodies were immobilized on the surface. The impedance was measured in consequence of the binding of bacteria with antibody on the electrodes surface. By fitting the impedance measurement into the parallel equivalent circuit, the resistance and capacitance of electrodes are obtained.

A rapid susceptibility test of Mycobacterium tuberculosis using hybridization protection assay

The conventional drug susceptibility tests of Mycobacterium tuberculosis are time-consuming and usually require 2 to 4 weeks to obtain final results. In this study, we attempted to develop a novel method for rapid detection of drug-resistant M. tuberculosis using hybridization protection assay (HPA). Clinically isolated strains of M. tuberculosis including seven isoniazid (INH) susceptible strains and six resistant strains were used. The organisms grown on Ogawa egg medium were inoculated into Middlebrook 7H9 broth and cultured at 37 degrees C for one week. Then, the inoculum of each strain was prepared as a tenfold dilution of bacterial suspension at McFarland No. 0.5. The inocula were mixed with INH solutions to yield final concentrations of 0.1 and 1.0 micrograms/ml, and the resulting bacterial suspensions with or without test drug were cultured on the swaying plate at 37 degrees C for up to 5 days. At intervals, 50 microliters of each sample was withdrawn and subjected to the protocol of the HPA using acridinium-ester (AE) labeled DNA probe, and then the relative light unit (RLU) was read in a luminometer. In the case of the susceptible strains, a significant difference in the mode of increase in RLU ratio (% of RLU on day x/RLU on day 0) was observed between the INH-treated and drug-free control sets within three days of cultivation, while no such differences were seen in the case of the resistant strains.(ABSTRACT TRUNCATED AT 250 WORDS)

A novel method for rapid detection of class IIa bacteriocin-producing lactic acid bacteria

A method based on colony PCR was developed for rapid screening of class IIa bacteriocin-producing lactic acid bacteria. The principle of the method is to detect the presence of bacteriocin gene by using specific degenerate primers to amplify a 3 kb fragment from bacteriocin structural gene to histidine kinase gene regions. Degenerate primers were designed according to the conserved amino acid sequences (YGNGVXCXXXXCXV and LDNAIE) contained in class IIa bacteriocins. This assay showed agreement with the conventional well-diffusion method. These findings indicate that the method proposed in this study is considerably more rapid and sufficient for screening of class IIa bacteriocin-producing bacterial isolates in environmental samples.

Optimization detection of plant pathogenic bacteria by electrochemiluminescence polymerase chain reaction method

Plant pathogenic bacteria spread all over the world, causing a great deal of economic loss. This study has developed a novel method for rapid detection of plant pathogenic bacteria by electrochemiluminescence polymerase chain reaction (ECL-PCR) using two universal probes, a biotin-probe and a Ru(bpy) 3 2+ (TBR)-probe. Biotin-probe sequence is complementary to the universal sequence of anti-sense primer, and TBR-probe sequence is the same as the universal sequence of sense primer. So the amplified PCR products can hybridize with TBR-probe and biotin-probe. After hybridizing, PCR products are captured by streptavidin coated magnetic bead through the biotin–streptavidin linkage, and then TBR reacts with tripropylamine to emit light for detection of plant pathogenic bacteria. This proposed method is applied to detect Fusarium oxysporum f. sp. Cubense and Xanthomonas oryzae pv. Oryzae. The results show that the method can successfully identify the plant pathogenic bacteria in the infected samples and these results are consistent with the results of gel electrophoresis. Importantly, this study can be used as an illustration for detecting various plant pathogenic bacteria and provides a feasible approach on developing ECL sensors to meet the demand of rapid detection of pathogenic bacteria, fungi, and viruses.

A novel method for rapid detection of Streptococcus pneumoniae antigen in sputum and its application in adult respiratory tract infections

A highly sensitive immunochromatography test kit, ODK0501, was developed using specific polyclonal antibodies against the C-polysaccharide moiety of Streptococcus pneumoniae for the rapid detection of S. pneumoniae antigen in sputum samples. The clinical utility of ODK0501 for this detection was evaluated prospectively in 52 adult patients with respiratory infections and compared with that of a urinary antigen detection kit. Overall, 21 patients (40.4 %) showed positive results with ODK0501, compared with 16 patients (30.8 %) using the urinary antigen detection kit, and S. pneumoniae was cultured from 18 patients. ODK0501 and the urinary antigen detection kit exhibited a sensitivity of 94.4 and 55.6 % (P<0.01), respectively, and a specificity of 88.2 and 82.4 %, respectively. Eleven of thirteen patients with conflicting results between the two test kits exhibited consistent results for sputum cultures. Moreover, eight out of nine patients positive for ODK0501 and negative for the urinary antigen detection kit were S. pneumoniae culture-positive, including five who exhibited phagocytosis, indicating S. pneumoniae as a causative agent of infection, in Gram staining of sputum samples. These results suggest that the ODK0501 direct sputum detection kit is more clinically useful than the urinary antigen detection kit in adult patients with respiratory infections.

Triple Helix Formation on Plasmid DNA Determined by a Size-Exclusion Chromatographic Method

Triple-helix-forming oligodeoxynucleotides are receiving considerable attention due to their potential applications for the inhibition of specific genesin vivo.However, their development is impaired by the lack of triple helix formation under physiological conditions. It is thus crucial to be able to quantitatively assay triple helix formation of various oligodeoxynucleotides on different target sequences. Usual methods to detect triple helix formation are restricted under the experimental conditions that can be studied. In addition, quantitative techniques are limited. We present a novel method for rapid detection and quantification of triple helix formation between an oligodeoxynucleotide and a plasmid carrying a target sequence. The oligodeoxynucleotide was radiolabeled and, after incubation with the target plasmid, the unbound oligodeoxynucleotide was separated from the mixture of plasmids and plasmid-bound oligodeoxynucleotides by rapid gel filtration spun columns. The formation of a triple helix between a target plasmid and several oligodeoxynucleotides was demonstrated and compared. Temperature, sequence and ionic dependencies, and kinetics of association were analyzed. This new technique can be used under a variety of conditions and should allow the rapid determination of optimal conditions required for triple helix formation, as well as the easy selection of an oligodeoxynucleotide that specifically binds with the highest affinity to a target double-stranded sequence.