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Rapid PCR

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https://www.readbyqxmd.com/read/29166662/detection-of-hbsag-mutants-in-the-blood-donor-population-of-pakistan
#1
Ahmad Farooq, Usman Waheed, Hasan Abbas Zaheer, Fahad Aldakheel, Shatha Alduraywish, Muhammad Arshad
BACKGROUND: Infection with the Hepatitis B virus (HBV) continues to be one of the leading healthcare issues in Pakistan, affecting over 6 million people. The existence of HBsAg mutants is well documented in many countries. In Pakistan, HBV screening in the majority of the blood banks is performed by Rapid Detection Devices or ELISA tests. These tests are designed to detect HBsAg, but may not detect the mutant HBsAg. Failure to detect the HBsAg mutant may result in the transmission of HBV infection from donor to recipient...
2017: PloS One
https://www.readbyqxmd.com/read/29166285/an-evaluation-of-the-usefulness-of-invasive-and-non-invasive-methods-used-to-diagnose-helicobacter-spp-infections-in-dogs
#2
M Jankowski, J Spużak, K Kubiak, K Glińska-Suchocka, M Biernat
The aim of this study was to assess the suitability of invasive and non-invasive methods used to diagnose Helicobacter spp. in the stomachs of dogs. The study was carried out on 30 dogs of both sexes and different breeds, between one and 15 years old. A histopathologic examination, a microbiological culture, a rapid urease test, a direct bacteriological preparation and a nested PCR assay were carried out. Gastric Helicobacter spp. was identified in gastric biopsy specimens from 16 (53.3%) dogs using direct bacteriological preparation, in four (13...
September 26, 2017: Polish Journal of Veterinary Sciences
https://www.readbyqxmd.com/read/29166280/differentiation-of-infectious-bronchitis-virus-vaccine-strains-ma5-and-4-91-by-taqman-real-time-pcr
#3
T Stenzel, D Dziewulska, M Śmiałek, B Tykałowski, J Kowalczyk, A Koncicki
The aim of this study was to develop rapid molecular assays for differentiating vaccine strains Ma5 and 4/91 of the infectious bronchitis virus (IBV). Specific primers and probes for S1 and N genes were designed based on the nucleotide sequences of both vaccine strains. Cross-reactivity was not observed. Assay sensitivity was 2.373 × 103 copies of the Ma5 strain, and 3.852 x 103 copies of the 4/91 strain. Samples belonging to a known genotype demonstrated that the designed assays supported rapid and sensitive detection of Ma5 and 4/91 vaccine strains of IBV...
September 26, 2017: Polish Journal of Veterinary Sciences
https://www.readbyqxmd.com/read/29166270/detection-of-yersinia-enterocolitica-species-in-pig-tonsils-and-raw-pork-meat-by-the-real-time-pcr-and-culture-methods
#4
M A Stachelska
The aim of the present study was to establish a rapid and accurate real-time PCR method to detect pathogenic Yersinia enterocolitica in pork. Yersinia enterocolitica is considered to be a crucial zoonosis, which can provoke diseases both in humans and animals. The classical culture methods designated to detect Y. enterocolitica species in food matrices are often very time-consuming. The chromosomal locus _tag CH49_3099 gene, that appears in pathogenic Y. enterocolitica strains, was applied as DNA target for the 5' nuclease PCR protocol...
September 26, 2017: Polish Journal of Veterinary Sciences
https://www.readbyqxmd.com/read/29163817/characteristics-and-genetic-diversity-of-multi-drug-resistant-extended-spectrum-beta-lactamase-esbl-producing-escherichia-coli-isolated-from-bovine-mastitis
#5
Tariq Ali, Sadeeq Ur Rahman, Limei Zhang, Muhammad Shahid, Dandan Han, Jian Gao, Shiyao Zhang, Pamela L Ruegg, Umer Saddique, Bo Han
A characterization of the drug resistance profiles, identification of PCR-based replicon typing, and multilocus sequence typing (MLST) and analysis of 46 ESBL-producing Escherichia coli from cows with mastitis are described. All multidrug-resistant isolates of various phylogenetic groups (A = 31, B1= 3, B2 = 2, D = 10) were ESBL-producers of genotypes CTX-M-15 (29), CTX-M-55 (4), CTX-M-14 (4), CTX-M-3 (1), CTX-M-1 (1), TEM (22) and SHV (8) that were found on conjugative plasmids of diverse incompatibility groups (primarily IncF)...
October 27, 2017: Oncotarget
https://www.readbyqxmd.com/read/29163647/comparison-of-diagnostic-tests-for-the-detection-of-bovine-brucellosis-in-the-natural-cases-of-abortion
#6
V Mahajan, H S Banga, G Filia, M P Gupta, K Gupta
Rapid and precise diagnosis in natural field cases of bovine abortion caused by Brucella abortus warrants the use of the most sensitive and reliable diagnostic methods. In the present study, bacterial isolation, serology, gross, histopathology, immunohistochemistry and polymerase chain reaction technique(s) were applied. Sero-prevalence studies showed the rate of 28.86% positive cases using the competitive ELISA. Histopathological changes were mainly seen in the placenta, fetal lungs, kidney, liver and spleen...
2017: Iranian journal of veterinary research
https://www.readbyqxmd.com/read/29163591/transcriptomic-and-hormonal-analyses-reveal-that-yuc-mediated-auxin-biogenesis-is-involved-in-shoot-regeneration-from-rhizome-in-cymbidium
#7
Yang Liu, Hai-Liang Zhang, He-Rong Guo, Li Xie, Rui-Zhen Zeng, Xiang-Qian Zhang, Zhi-Sheng Zhang
Cymbidium, one of the most important orchid genera in horticulture, can be classified into epiphytic and terrestrial species. Generally, epiphytic Cymbidium seedlings can be easily propagated by tissue culture, but terrestrial seedlings are difficult to propagate. To date, the molecular mechanisms underlying the differences in the ease with which terrestrial and epiphytic cymbidiums can be propagated are largely unknown. Using RNA-sequencing, quantitative reverse transcription PCR and enzyme-linked immunosorbent assay, Cymbidium 'Xiaofeng' (CXF), which can be efficiently micropropagated, and terrestrial Cymbidium sinense 'Qijianbaimo' (CSQ), which has a low regeneration ability, were used to explore the molecular mechanisms underlying the micropropagation ability of Cymbidium species...
2017: Frontiers in Plant Science
https://www.readbyqxmd.com/read/29163484/tracking-human-immunodeficiency-virus-1-infection-in-the-humanized-drag-mouse-model
#8
Jiae Kim, Kristina K Peachman, Ousman Jobe, Elaine B Morrison, Atef Allam, Linda Jagodzinski, Sofia A Casares, Mangala Rao
Humanized mice are emerging as an alternative model system to well-established non-human primate (NHP) models for studying human immunodeficiency virus (HIV)-1 biology and pathogenesis. Although both NHP and humanized mice have their own strengths and could never truly reflect the complex human immune system and biology, there are several advantages of using the humanized mice in terms of using primary HIV-1 for infection instead of simian immunodeficiency virus or chimera simian/HIV. Several different types of humanized mice have been developed with varying levels of reconstitution of human CD45(+) cells...
2017: Frontiers in Immunology
https://www.readbyqxmd.com/read/29163387/a-multiplex-sybr-green-real-time-pcr-assay-for-the-detection-of-three-colistin-resistance-genes-from-cultured-bacteria-feces-and-environment-samples
#9
Jiyun Li, Xiaomin Shi, Wenjuan Yin, Yang Wang, Zhangqi Shen, Shuangyang Ding, Shaolin Wang
The aim of the study was to develop a multiplex assay for rapid detection of mcr-1, mcr-2, and mcr-3, a group of genes of conferring resistance to colistin mediated by plasmid in Enterobacteriaceae. A SYBR Green based real-time PCR assay has been designed to detect the mcr genes, and applied to cultured bacteria, feces and soil samples. All three mcr genes could be detected with a lower limit of 10(2) cultured bacteria. This test was highly specific and sensitive, and generated no false-positive results. The assay was also conclusive when applied to feces and soil samples containing mcr-1-positive Escherichia coli, which could facilitate the screening of mcr genes not only in the bacteria, but also directly from the environment...
2017: Frontiers in Microbiology
https://www.readbyqxmd.com/read/29160018/multi-fragment-melting-analysis-of-yeast-species-isolated-from-spoiled-fruits
#10
Zülal Kesmen, Esra Özbekar, Mine Erdem Büyükkiraz
AIMS: In this study, we developed a novel approach, multi-fragment melting analysis (MFMA), based on the simultaneous analysis of the melting characteristics of multiple DNA fragments for the differentiation and identification of yeast species isolated from spoiled fruits. METHODS AND RESULTS: In total, 183 yeast isolates recovered from spoiled fruit samples were differentiated and grouped using MFMA. Six different DNA fragments of the 26S rRNA gene showing a high interspecific heterogeneity were amplified, and the PCR products were individually subjected to MFMA...
November 21, 2017: Journal of Applied Microbiology
https://www.readbyqxmd.com/read/29158131/development-of-a-low-cost-paper-based-elisa-method-for-rapid-escherichia-coli-o157-h7-detection
#11
REVIEW
Bo Pang, Chao Zhao, Li Li, Xiuling Song, Kun Xu, Juan Wang, Yushen Liu, Kaiyue Fu, Hao Bao, Dandan Song, Xiangjun Meng, Xiaofeng Qu, Zhuping Zhang, Juan Li
Escherichia coli O157: H7 (E. coli O157: H7) has become one of the most dangerous foodborne pathogenic bacteria around the world. Currently, because of the tedious, high-cost and stringent laboratory conditions required, the conventional E. coli O157: H7 detection methods, such as culture-based methods and polymerase chain reaction (PCR), have much limitation. Thus, we developed a novel paper-based enzyme-linked immunosorbent assay (p-ELISA) with shorter operation duration, lower cost, relatively higher sensitivity and wider application...
November 17, 2017: Analytical Biochemistry
https://www.readbyqxmd.com/read/29156877/comparison-of-microscopy-elisa-and-real-time-pcr-for-detection-of-giardia-intestinalis-in-human-stool-specimens
#12
Yunus Emre Beyhan, Zeynep Taş Cengiz
Background/aim: This study included patients who had digestive system complaints between August 2015 and October 2015. The research was designed to compare conventional microscopy with an antigen detection ELISA kit and the TaqMan-based real-time PCR (RT-PCR) technique for detection of Giardia intestinalis in human stool specimens. Materials and methods: Samples were concentrated by formalin-ether sedimentation technique and microscopic examinations were carried out on wet mount slides. A commercially available ELISA kit (Giardia CELISA, Cellabs, Brookvale, Australia) was used for immunoassay...
August 23, 2017: Turkish Journal of Medical Sciences
https://www.readbyqxmd.com/read/29155729/sample-preparation-and-analysis-of-rnaseq-based-gene-expression-data-from-zebrafish
#13
Timothy L Hostelley, Jessica E Nesmith, Norann A Zaghloul
The analysis of global gene expression changes is a valuable tool for identifying novel pathways underlying observed phenotypes. The zebrafish is an excellent model for rapid assessment of whole transcriptome from whole animal or individual cell populations due to the ease of isolation of RNA from large numbers of animals. Here a protocol for global gene expression analysis in zebrafish embryos using RNA sequencing (RNASeq) is presented. We describe preparation of RNA from whole embryos or from cell populations obtained using cell sorting in transgenic animals...
October 27, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/29155724/generation-of-a-gene-disrupted-streptococcus-mutans-strain-without-gene-cloning
#14
Takatoshi Murata, Ayako Okada, Khairul Matin, Nobuhiro Hanada
Typical methods for the elucidation of the function of a particular gene involve comparative phenotypic analyses of the wild-type strain and a strain in which the gene of interest has been disrupted. A gene-disruption DNA construct containing a suitable antibiotic resistance marker gene is useful for the generation of gene-disrupted strains in bacteria. However, conventional construction methods, which require gene cloning steps, involve complex and time-consuming protocols. Here, a relatively facile, rapid, and cost-effective method for targeted gene disruption in Streptococcus mutans is described...
October 23, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/29155238/evaluation-of-two-real-time-pcr-assays-for-the-detection-of-bacterial-dna-in-amniotic-fluid
#15
Patricia Girón de Velasco-Sada, Iker Falces-Romero, Inmaculada Quiles-Melero, Adela García-Perea, Jesús Mingorance
PURPOSE: The aim of this study was to evaluate two non-commercial Real-Time PCR assays for the detection of microorganisms in amniotic fluid followed by identification by pyrosequencing. METHODS: We collected 126 amniotic fluids from 2010 to 2015 for the evaluation of two Real-Time PCR assays for detection of bacterial DNA in amniotic fluid (16S Universal PCR and Ureaplasma spp. specific PCR). The method was developed in the Department of Microbiology of the University Hospital La Paz...
November 16, 2017: Journal of Microbiological Methods
https://www.readbyqxmd.com/read/29154392/rapid-and-simple-detection-of-ureaplasma-species-from-vaginal-swab-samples-using-a-loop-mediated-isothermal-amplification-method
#16
Kazumasa Fuwa, Mitsuko Seki, Yoshiyasu Hirata, Itaru Yanagihara, Yukiko Nakura, Chika Takano, Kazumichi Kuroda, Satoshi Hayakawa
PROBLEM: Ureaplasma species occasionally cause chorioamnionitis and premature labor. We developed a novel assay employing a loop-mediated isothermal amplification (LAMP) method to detect Ureaplasma parvum and Ureaplasma urealyticum. METHOD OF STUDY: Loop-mediated isothermal amplification primers were designed to amplify Ureaplasma-specific ureaseB genes. Four U. parvum strains, 5 U. urealyticum strains and 14 reference bacterial species were evaluated. Forty-six vaginal swab samples were analyzed by LAMP, culture, and PCR...
November 20, 2017: American Journal of Reproductive Immunology: AJRI
https://www.readbyqxmd.com/read/29153517/rapid-species-and-subspecies-specific-level-classification-and-identification-of-lactobacillus-casei-group-members-using-maldi-biotyper-combined-with-clinprotools
#17
Chien-Hsun Huang, Lina Huang
Using common taxonomic methods such as 16S rDNA sequencing and physiological and biochemical analysis to identify members of the Lactobacillus casei group (LCG) is time-consuming, expensive and inaccurate. In this study, we applied matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) to rapid discriminate LCG strains by creating an analytical in-house database (IHDB) and to develop a classification model for subspecies-level differentiation based on MS biomarkers using ClinProTools bioinformatics software (Bruker Daltonics, Billerica, MA)...
November 15, 2017: Journal of Dairy Science
https://www.readbyqxmd.com/read/29152132/high-baalc-copy-numbers-in-peripheral-blood-prior-to-allogeneic-transplantation-predict-early-relapse-in-acute-myeloid-leukemia-patients
#18
Madlen Jentzsch, Marius Bill, Juliane Grimm, Julia Schulz, Karoline Goldmann, Stefanie Beinicke, Janine Häntschel, Wolfram Pönisch, Georg-Nikolaus Franke, Vladan Vucinic, Gerhard Behre, Thoralf Lange, Dietger Niederwieser, Sebastian Schwind
High BAALC expression levels at acute myeloid leukemia diagnosis have been linked to adverse outcomes. Recent data indicate that high BAALC expression levels may also be used as marker for residual disease following acute myeloid leukemia treatment. Allogeneic hematopoietic stem cell transplantation (HSCT) offers a curative treatment for acute myeloid leukemia patients. However, disease recurrence remains a major clinical challenge and identification of high-risk patients prior to HSCT is crucial to improve outcomes...
October 20, 2017: Oncotarget
https://www.readbyqxmd.com/read/29151962/insufficient-radiofrequency-ablation-treated-hepatocellular-carcinoma-cells-promote-metastasis-by-up-regulation-itgb3
#19
Ning Zhang, Dening Ma, Lu Wang, Xiaodong Zhu, Qi Pan, Yiming Zhao, Weiping Zhu, Jiamin Zhou, Longrong Wang, Zongtao Chai, Jianyang Ao, Huichuan Sun, Zhaoyou Tang
Radiofrequency ablation (RFA) is one of the standards of care for early stage hepatocellular carcinoma (HCC). However, rapid progression of residual tumor after RFA has been confirmed. The aim of this study was to investigate the underlying mechanism of this phenomenon. Human HCC cell lines HCCLM3 and HepG2 were employed to establish insufficient RFA models in vivo and in vitro, respectively. The effects of insufficient RFA on metastatic potential of residual tumors were evaluated. The molecular changes after insufficient RFA were evaluated by PCR array, western blot, immunofluorescence, and immunohistochemistry...
2017: Journal of Cancer
https://www.readbyqxmd.com/read/29151824/a-real-time-metabolomic-profiling-approach-to-detecting-fish-fraud-using-rapid-evaporative-ionisation-mass-spectrometry
#20
Connor Black, Olivier P Chevallier, Simon A Haughey, Julia Balog, Sara Stead, Steven D Pringle, Maria V Riina, Francesca Martucci, Pier L Acutis, Mike Morris, Dimitrios S Nikolopoulos, Zoltan Takats, Christopher T Elliott
Introduction: Fish fraud detection is mainly carried out using a genomic profiling approach requiring long and complex sample preparations and assay running times. Rapid evaporative ionisation mass spectrometry (REIMS) can circumvent these issues without sacrificing a loss in the quality of results. Objectives: To demonstrate that REIMS can be used as a fast profiling technique capable of achieving accurate species identification without the need for any sample preparation...
2017: Metabolomics: Official Journal of the Metabolomic Society
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