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Rapid PCR

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https://www.readbyqxmd.com/read/29356220/comparison-of-false-negative-rates-and-limits-of-detection-following-macrofoam-swab-sampling-of-bacillus-anthracis-surrogates-via-rapid-viability-pcr-and-plate-culture
#1
Janine R Hutchison, Greg F Piepel, Brett G Amidan, Becky M Hess, Michael A Sydor, Brooke L Deatherage Kaiser
AIMS: We evaluated the effects of Bacillus anthracis surrogates, low surface concentrations, surface materials, and assay methods on false-negative rate (FNR) and limit of detection (LOD95 ) for recovering Bacillus spores using a macrofoam-swab sampling procedure. METHODS AND RESULTS: Bacillus anthracis Sterne or Bacillus atrophaeus Nakamura spores were deposited over a range of low target concentrations (2 - 500 coupon-1 ) onto glass, stainless steel, vinyl tile, and plastic...
January 21, 2018: Journal of Applied Microbiology
https://www.readbyqxmd.com/read/29355148/multiplex-polymerase-chain-reaction-of-genetic-markers-for-detection-of-potentially-pathogenic-environmental-legionella-pneumophila-isolates
#2
Arvind Valavane, Rama Chaudhry, Pawan Malhotra
BACKGROUND & OBJECTIVES: Genomic constitution of the bacterium Legionella pneumophila plays an important role in providing them a pathogenic potential. Here, we report the standardization and application of multiplex polymerase chain reaction (PCR) for the detection of molecular markers of pathogenic potential in L. pneumophila in hospital environment. METHODS: Culture of the standard strains of L. pneumophila was performed in buffered charcoal-yeast extract agar with L-cysteine at p H 6...
September 2017: Indian Journal of Medical Research
https://www.readbyqxmd.com/read/29353377/highly-synergistic-activity-of-melittin-with-imipenem-and-colistin-in-biofilm-inhibition-against-multidrug-resistant-strong-biofilm-producer-strains-of-acinetobacter-baumannii
#3
Ali Mohammadi Bardbari, Mohammad Reza Arabestani, Manoochehr Karami, Fariba Keramat, Hossein Aghazadeh, Mohammad Yousef Alikhani, Kamran Pooshang Bagheri
The rapid increase of drug resistance and failure of available antibiotics to treat biofilm-associated infections is of great health concern. Accordingly, our study aimed to evaluate the synergistic antibacterial, biofilm inhibitory, and biofilm removal activities of melittin in combination with colistin, imipenem, and ciprofloxacin against multidrug-resistant (MDR) strong biofilm producer Acinetobacter baumannii isolates. The kinetics of biofilm formation were evaluated for the isolates for 144 h. Minimum inhibitory concentrations (MICs), minimum bactericidal concentrations (MBCs), minimum biofilm inhibitory concentrations (MBICs), and biofilm removal activities for melittin and combinations with antibiotics were determined...
January 20, 2018: European Journal of Clinical Microbiology & Infectious Diseases
https://www.readbyqxmd.com/read/29353007/nitric-oxide-suppresses-aluminum-induced-programmed-cell-death-in-peanut-arachis-hypoganea-l-root-tips-by-improving-mitochondrial-physiological-properties
#4
Huyi He, Wenjing Huang, Thet Lwin Oo, Minghua Gu, Jie Zhan, Aiqin Wang, Long-Fei He
Aluminum (Al) stress alters nitric oxide (NO) and induces programmed cell death (PCD) in plants. Recent study has shown that NO inhibits Al-induced PCD. However, the mechanism of NO inhibiting Al-induced PCD has not been revealed yet. Here, we investigated the behavior of mitochondria during Al-induced PCD suppressed by NO in peanut. Seedlings of peanut was grown hydroponically in a controllable growth room. The mitochondrial physiological parameters were determined spectrophotometrically. The expression of AhANT and AhHsp70 was determined by quantitative RT-PCR...
January 15, 2018: Nitric Oxide: Biology and Chemistry
https://www.readbyqxmd.com/read/29352704/treatment-of-adult-chronic-indeterminate-chagas-disease-with-benznidazole-and-three-e1224-dosing-regimens-a-proof-of-concept-randomised-placebo-controlled-trial
#5
Faustino Torrico, Joaquim Gascon, Lourdes Ortiz, Cristina Alonso-Vega, María-Jesús Pinazo, Alejandro Schijman, Igor C Almeida, Fabiana Alves, Nathalie Strub-Wourgaft, Isabela Ribeiro
BACKGROUND: Chagas disease is a major neglected vector-borne disease. In this study, we investigated the safety and efficacy of three oral E1224 (a water-soluble ravuconazole prodrug) regimens and benznidazole versus placebo in adult chronic indeterminate Chagas disease. METHOD: In this proof-of-concept, double-blind, randomised phase 2 clinical trial, we recruited adults (18-50 years) with confirmed diagnosis of Trypanosoma cruzi infection from two outpatient units in Bolivia...
January 15, 2018: Lancet Infectious Diseases
https://www.readbyqxmd.com/read/29351668/evaluation-of-the-rapid-carbapenem-inactivation-method-rcim-a-phenotypic-screening-test-for-carbapenemase-producing-enterobacteriaceae
#6
Madalina-Maria Muntean, Andrei-Alexandru Muntean, Lauraine Gauthier, Elodie Creton, Garance Cotellon, Mircea Ioan Popa, Rémy A Bonnin, Thierry Naas
Objectives: Fast and accurate diagnostic tests to identify carbapenemase-producing Enterobacteriaceae (CPE) are mandatory for proper antimicrobial therapy and implementing infection control measures. Here, we have developed a rapid Carbapenem Inactivation Method (rCIM) for CPE detection. Methods: The rCIM consists of the incubation of a potential carbapenemase producer with meropenem discs and use of the resulting supernatant to challenge a susceptible indicator strain...
January 16, 2018: Journal of Antimicrobial Chemotherapy
https://www.readbyqxmd.com/read/29351385/development-of-a-recombinase-polymerase-amplification-assay-for-vibrio-parahaemolyticus-detection-with-an-internal-amplification-control
#7
Huanlan Yang, Shuang Wei, Ravi Gooneratne, Anthony N Mutukumirad, Xuejun Ma, Shuze Tang, Xiyang Wu
A novel RPA-IAC assay using recombinase polymerase and an internal amplification control (IAC) for Vibrio parahaemolyticus detection was developed. Specific primers were designed based on the coding sequence (CDS) for the toxR gene in V. parahaemolyticus. The recombinase polymerase amplification (RPA) reaction was conducted at a constant low temperature of 37°C for 20 min. Assay specificity was validated by using 63 Vibrio strains and 10 non-Vibrio bacterial species. In addition, a competitive IAC was employed to avoid false-negative results, which co-amplified simultaneously with the target sequence...
January 19, 2018: Canadian Journal of Microbiology
https://www.readbyqxmd.com/read/29351317/hafts-are-novel-lncrna-transcripts-from-aflatoxin-exposure
#8
B Alex Merrick, Justin S Chang, Dhiral P Phadke, Meredith A Bostrom, Ruchir R Shah, Xinguo Wang, Oksana Gordon, Garron M Wright
The transcriptome can reveal insights into precancer biology. We recently conducted RNA-Seq analysis on liver RNA from male rats exposed to the carcinogen, aflatoxin B1 (AFB1), for 90 days prior to liver tumor onset. Among >1,000 differentially expressed transcripts, several novel, unannotated Cufflinks-assembled transcripts, or HAfTs (Hepatic Aflatoxin Transcripts) were found. We hypothesized PCR-cloning and RACE (rapid amplification of cDNA ends) could further HAfT identification. Sanger data was obtained for 6 transcripts by PCR and 16 transcripts by 5'- and 3'-RACE...
2018: PloS One
https://www.readbyqxmd.com/read/29350545/sensitive-multiplex-real-time-rt-qpcr-assay-for-the-detection-of-filoviruses
#9
Vladimir G Dedkov, N'Faly Magassouba, Marina V Safonova, Sergey A Bodnev, Oleg V Pyankov, Jacob Camara, Bakary Sylla, Alexander P Agafonov, Victor V Maleev, German A Shipulin
Filoviruses are important etiological agents of emergent diseases with high mortality rates. Traditionally, filovirus fever diseases have primarily been a burden of African countries; however, global interconnectedness has increased the probability of the worldwide spread of filoviruses. Therefore, national healthcare organizations need tools for managing filovirus risk, including diagnostic kits based on real-time reverse transcription PCR (RT-qPCR), as this is the most suitable method for diagnosing filovirus fever diseases...
January 19, 2018: Health Security
https://www.readbyqxmd.com/read/29350529/a-colorimetric-dnazyme-biosensor-for-convenience-detection-of-enterotoxin-b-harbouring-staphylococcus-aureus-from-food-samples
#10
Bhairab Mondal, Bhavanashri N, Shylaja Ramlal, Joseph Kingston
In the present study, a colorimetric DNAzymes biosensor strategy was devised in combination with immunomagnetic separation for rapid and easy detection of enterotoxin B harbouring Staphylococcus aureus from food and clinical samples. The method employs immunocapture of S. aureus and amplification of seb gene by DNAzyme complementary sequence integrated forward primer and with specific reverse primer. The DNAzyme sequence integrated dsDNA PCR products when treated with hemin and TMB (3, 3', 5, 5'-tetramethylbenzidine) in the presence of H2O2 produce colorimetric signal...
January 19, 2018: Journal of Agricultural and Food Chemistry
https://www.readbyqxmd.com/read/29350524/development-of-a-multiplexed-microsphere-pcr-for-rapid-culture-free-detection-and-gram-typing-of-bacteria-in-human-blood-samples
#11
Fang Liang, Daniel J Browne, Megan J Gray, Kate H Gartlan, David D Smith, Ross Barnard, Geoff R Hill, Simon Robert Corrie, Kate Markey
Blood stream infection is a significant clinical problem, particularly in vulnerable patient groups such as those undergoing chemotherapy and bone marrow transplantation. Clinical diagnostics for suspected blood stream infection remain centered around blood culture (highly variable timing, hours to days), and empiric use of broad-spectrum antibiotics is often employed for patients presenting with febrile neutropenia. Gram-typing provides the first opportunity to target therapy (e.g. combinations containing vancomycin or teicoplanin for Gram-positives; piperacillin-tazobactam or a carbapenem for Gram-negatives), however current approaches require blood culture...
January 19, 2018: ACS Infectious Diseases
https://www.readbyqxmd.com/read/29350442/role-of-ethylene-and-related-gene-expression-in-the-interaction-between-strawberry-plants-and-the-plant-growth-promoting-bacterium-azospirillum-brasilense
#12
J M Elías, M F Guerrero-Molina, M G Martínez-Zamora, J C Díaz-Ricci, R O Pedraza
• The induced systemic resistance (ISR) is one of the indirect mechanisms of growth promotion exerted by plant growth-promoting bacteria, and can be mediated by ethylene (ET). We assessed the ET production and the expression of related genes in the Azospirillum-strawberry plant interaction. • ET production was evaluated by gas chromatography in plants inoculated or not with A. brasilense REC3. Also, plants were treated with AgNO3 , an inhibitor of ET biosynthesis; with 1-aminocyclopropane-1-carboxylic-acid (ACC), a precursor of ET biosynthesis; and with indole acetic acid (IAA)...
January 19, 2018: Plant Biology
https://www.readbyqxmd.com/read/29350441/ultrasensitive-detection-of-shiga-toxin-2-and-its-variants-in-shiga-toxin-producing-escherichia-coli-strains-by-a-time-resolved-fluorescence-immunoassay
#13
Tian Wen, Chao Huang, Yi Zhang, Xiaoyan Zeng, Wendong Liu, Zhenbang Jiao, Xiling Guo, Yongjun Jiao
A rapid and sensitive two-step time-resolved fluorescence immunoassay (TRFIA) was developed for the detection of Shiga toxin 2 (Stx2) and its variants in Shiga toxin-producing Escherichia coli (STEC) strains. In sandwich mode, a monoclonal antibody against Stx2 was coated on a microtiter plate as a capture antibody. A tracer antibody against Stx2 labeled with europium(III) (Eu3+ ) chelate was then used as a detector, followed by fluorescence measurements using time-resolved fluorescence. The sensitivity of Stx2 detection was 0...
January 19, 2018: Luminescence: the Journal of Biological and Chemical Luminescence
https://www.readbyqxmd.com/read/29350029/a-microwell-array-method-for-rapid-generation-of-uniform-agarose-droplets-and-beads-for-single-molecule-analysis
#14
Xingrui Li, Dongfeng Zhang, Huimin Zhang, Zhichao Guan, Yanling Song, Ruochen Liu, Zhi Zhu, Chaoyong James Yang
Compartmentalization of aqueous samples in uniform emulsion droplets has proven to be a useful tool for many chemical, biological, and biomedical applications. Herein, we introduce an array-based emulsification method for rapid and easy generation of monodisperse agarose-in-oil droplets in a PDMS microwell array. The microwells are filled with agarose solution, and subsequent addition of hot oil results in immediate formation of agarose droplets due to the surface-tension of the liquid solution. Because droplet size is determined solely by the array unit dimensions, uniform droplets with pre-selectable diameters ranging from 20 µm to 100 µm can be produced with relative standard deviations less than 3...
January 19, 2018: Analytical Chemistry
https://www.readbyqxmd.com/read/29349550/recombinase-polymerase-amplification-combined-with-lateral-flow-dipstick-for-equipment-free-detection-of-salmonella-in-shellfish
#15
Weifang Gao, Hailong Huang, Peng Zhu, Xiaojun Yan, Jianzhong Fan, Jinpo Jiang, Jilin Xu
Salmonella is a major pathogen that causes acute foodborne outbreaks worldwide. Seafood, particularly shellfish, is a proven source of Salmonella spp. infection because many people prefer to eat it raw or lightly cooked. However, traditional identification methods are too time-consuming and complex to detect contamination of bacteria in the food chain in a timely manner, and few studies have aimed to identify Salmonella in shellfish early in the supply chain. We herein developed a method for rapid detection of Salmonella in shellfish based on the method of recombinase polymerase amplification (RPA) combined with lateral flow dipstick (LFD), which targets the invasion gene A (invA)...
January 18, 2018: Bioprocess and Biosystems Engineering
https://www.readbyqxmd.com/read/29346412/a-comparative-analysis-of-preservation-techniques-for-the-optimal-molecular-detection-of-hookworm-dna-in-a-human-fecal-specimen
#16
Marina Papaiakovou, Nils Pilotte, Ben Baumer, Jessica Grant, Kristjana Asbjornsdottir, Fabian Schaer, Yan Hu, Raffi Aroian, Judd Walson, Steven A Williams
BACKGROUND: Proper collection and storage of fecal samples is necessary to guarantee the subsequent reliability of DNA-based soil-transmitted helminth diagnostic procedures. Previous research has examined various methods to preserve fecal samples for subsequent microscopic analysis or for subsequent determination of overall DNA yields obtained following DNA extraction. However, only limited research has focused on the preservation of soil-transmitted helminth DNA in stool samples stored at ambient temperature or maintained in a cold chain for extended periods of time...
January 2018: PLoS Neglected Tropical Diseases
https://www.readbyqxmd.com/read/29346384/histoplasmosis-in-africa-an-emerging-or-a-neglected-disease
#17
REVIEW
Rita O Oladele, Olusola O Ayanlowo, Malcolm D Richardson, David W Denning
Histoplasmosis in Africa has markedly increased since the advent of the HIV/AIDS epidemic but is under-recognised. Pulmonary histoplasmosis may be misdiagnosed as tuberculosis (TB). In the last six decades (1952-2017), 470 cases of histoplasmosis have been reported. HIV-infected patients accounted for 38% (178) of the cases. West Africa had the highest number of recorded cases with 179; the majority (162 cases) were caused by Histoplasma capsulatum var. dubuosii (Hcd). From the Southern African region, 150 cases have been reported, and the majority (119) were caused by H...
January 2018: PLoS Neglected Tropical Diseases
https://www.readbyqxmd.com/read/29344852/laser-capture-microdissection-insights-into-methods-and-applications
#18
Meera Mahalingam
Laser capture microdissection is a non-molecular, minimally disruptive method to obtain cytologically and/or phenotypically defined cells or groups of cells from heterogeneous tissues. Its advantages include efficient rapid and precise procurement of cells. The potential disadvantages include time consuming, expensive, and limited by the need for a pathologist for recognition of distinct subpopulations within a specified sample. Overall it is versatile allowing the preparation of homogenous isolates of specific subpopulations of cells from which DNA/RNA or protein can be extracted for RT-PCR, quantitative PCR, next-generation sequencing, immunoblot blot analyses, and mass spectrometry...
2018: Methods in Molecular Biology
https://www.readbyqxmd.com/read/29343880/novel-application-of-loop-mediated-isothermal-amplification-for-rapid-detection-of-gene-translocation
#19
Ibu Matsuzaki, Hideto Iguchi, Yurina Mikasa, Hiromu Morishita, Katsuya Okuda, Keita Nakaguchi, Yuki Mori, Yoshifumi Iwahashi, Kenji Warigaya, Masakazu Fujimoto, Fumiyoshi Kojima, Shin-Ichi Murata
Identification of fusion genes in cancer is essential for pathological diagnosis and clinical therapy. Although methods for detection of fusion genes, such as fluorescence in situ hybridization (FISH) and real-time polymerase chain reaction (PCR), have been developed in last two decades, these methods are not ideal for detection of these genetic alterations owing to their high cost and time-consuming procedures. In this study, we developed novel application for detection of gene translocations using loop-mediated isothermal amplification (LAMP)...
December 26, 2017: Acta Histochemica et Cytochemica
https://www.readbyqxmd.com/read/29342162/comparison-of-diagnostic-methods-to-detect-histoplasma-capsulatum-in-serum-and-blood-samples-from-aids-patients
#20
Katia Cristina Dantas, Roseli Santos de Freitas, Marcos Vinicius da Silva, Paulo Ricardo Criado, Olinda do Carmo Luiz, Adriana Pardini Vicentini
BACKGROUND: Although early and rapid detection of histoplasmosis is essential to prevent morbidity and mortality, few diagnostic tools are available in resource-limited areas, especially where it is endemic and HIV/AIDS is also epidemic. Thus, we compared conventional and molecular methods to detect Histoplasma capsulatum in sera and blood from HIV/AIDS patients. METHODOLOGY: We collected a total of 40 samples from control volunteers and patients suspected of histoplasmosis, some of whom were also infected with other pathogens...
2018: PloS One
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