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Molecular and Cellular Probes

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https://www.readbyqxmd.com/read/28958719/recombinase-polymerase-amplification-assay-for-rapid-detection-of-porcine-circovirus-3
#1
Jianchang Wang, Yongning Zhang, Ruoxi Zhang, Qingan Han, Jinfeng Wang, Libing Liu, Ruiwen Li, Wanzhe Yuan
The objective of this study was to develop a real-time recombinase polymerase amplification (rt-RPA) assay for the rapid detection of porcine circovirus 3 (PCV3). Specific RPA primers and exo probes were designed for the cap gene of PCV3 within the conserved region of viral genome. The amplification was performed at 38 °C for 20 min. The rt-RPA was specific for PCV3, as there was no cross-reaction with other pathogens tested. Using the recombinant plasmid pUC57-PCV3 as template, the analytical sensitivity was 23 copies...
September 25, 2017: Molecular and Cellular Probes
https://www.readbyqxmd.com/read/28863892/simultaneous-detection-and-differentiation-of-human-parvovirus-b19-and-human-parvovirus-4-by-an-internally-controlled-multiplex-quantitative-real-time-pcr
#2
Junting Jia, Yadi Zhong, Yi Guo, Chaoji Huangfu, Xiong Zhao, Chi Fang, Rui Fan, Yuyuan Ma, Jingang Zhang
Human parvovirus B19 (B19V) and human parvovirus 4 (PARV4) are two parvoviruses known to infect humans and transmit through blood and plasma derived medicinal products (PDMPs). Inactivation of the two parvoviruses has proven to be difficult and nucleic acid testing (NAT) would be an efficient means to exclude viruses. In this study, an internally controlled multiplex quantitative real-time PCR (qPCR) assay for B19V and PARV4 simultaneous detection and quantification was established and evaluated. The optimized multiplex qPCR assay allowed for simultaneous detection of all of the genotypes (1-3) of B19V and PARV4, with equal limit of quantification (LOQ) of 5 copies/μL, rather than other blood-borne viruses...
August 30, 2017: Molecular and Cellular Probes
https://www.readbyqxmd.com/read/28842221/rapid-and-visual-detection-of-mycobacterium-tuberculosis-complex-using-recombinase-polymerase-amplification-combined-with-lateral-flow-strips
#3
Qinglin Ma, Houming Liu, Feidi Ye, Guangxin Xiang, Wanshui Shan, Wanli Xing
To definitively diagnose active pulmonary Tuberculosis (TB), Mycobacterium tuberculosis complex (MTBC) bacilli must be identified within clinical specimens from patients. In this study, we introduced a rapid and visual detection method of MTBC using recombinase polymerase amplification (RPA) combined with lateral flow (LF) strips. The LF-RPA assay, read results with naked eyes, could detect as few as 5 genome copies of M. tuberculosis H37Rv (ATCC 27294) per reaction and had no cross-reactions with other control bacteria even using excessive amount of template DNA...
August 22, 2017: Molecular and Cellular Probes
https://www.readbyqxmd.com/read/28826997/detection-of-skeletonema-costatum-based-on-loop-mediated-isothermal-ampli%C3%AF-cation-combined-with-lateral-%C3%AF-ow-dipstick
#4
Hai-Long Huang, Peng Zhu, Cheng-Xu Zhou, Xiao-Jun Yan, Yi-Xin Zou, Pei-Wen Lv
We developed a new assay method, which combines loop-mediated isothermal amplification (LAMP) with a chromatographic lateral flow dipstick (LFD) for the rapid and special detection of the diatom Skeletonema costatum. Four groups of LAMP primers were derived from a conserved DNA sequence unique to S. costatum. The amplifications were carried out at 61, 63, and 65 °C for 60 min in various combinations by the quantitative PCR thermal cycler to confirm optimal primers and reaction temperature. The LAMP-LFD detection limit was 0...
August 18, 2017: Molecular and Cellular Probes
https://www.readbyqxmd.com/read/28823562/identification-and-characterization-of-adhesion-proteins-in-lactobacilli-targeting-actin-as-receptor
#5
Zhen Peng, Rudi F Vogel, Matthias A Ehrmann, Tao Xiong
Actin as the main constitution of cytoskeleton in host cells plays an important role in mediating bacterial colonization. To identify the actin-binding proteins in Lactobacillus (L.) paracasei, L. plantarum, and L. brevis, actin immobilized to 24-well plate was used to probe adhesion proteins. Five adhesion proteins were identified and characterized by electrophoresis and LC-MS/MS: pyruvate kinase (PK), glucose-6-phosphate isomerase (PGI), phosphoglycerate kinase (PGK), chaperonin GroEL, and EF-Tu, all of which could display on the cell surface, indicating their possible role in mediating bacterial adhesion to host...
August 17, 2017: Molecular and Cellular Probes
https://www.readbyqxmd.com/read/28803792/rapid-and-visual-detection-of-leptospira-in-urine-by-ligb-lamp-assay-with-pre-addition-of-dye
#6
Syed Atif Ali, Gurpreet Kaur, Nongthombam Boby, T Sabarinath, Khushal Solanki, Dheeraj Pal, Pallab Chaudhuri
Leptospirosis is considered to be the most widespread zoonotic disease caused by pathogenic species of Leptospira. The present study reports a novel set of primers targeting LigB gene for visual detection of pathogenic Leptospira in urine samples through Loop-mediated isothermal amplification (LAMP). The results were recorded by using Hydroxyl napthol blue (HNB), SYBR GREEN I and calcein. Analytical sensitivity of LAMP was as few as 10 leptospiral organisms in spiked urine samples from cattle and dog. LigB gene based LAMP, termed as LigB-LAMP, was found 10 times more sensitive than conventional PCR...
August 10, 2017: Molecular and Cellular Probes
https://www.readbyqxmd.com/read/28652020/development-of-a-realtime-rt-pcr-assay-for-the-rapid-detection-of-influenza-a-h2-viruses
#7
Andrey Komissarov, Artem Fadeev, Anna Kosheleva, Kseniya Sintsova, Mikhail Grudinin
Influenza and other acute respiratory infections are of great concern for public health, causing excessive morbidity and mortality throughout the world. Influenza virus A(H2N2), which caused a pandemic of so called "Asian flu" in 1957 was expelled from the human population by the new pandemic virus subtype H3N2 in 1968, however, influenza A(H2) viruses continue to circulate in wild birds and poultry. The lack of immunity in human population and the continued circulation of influenza A(H2) among animals makes emergence of a new pandemic virus possible...
October 2017: Molecular and Cellular Probes
https://www.readbyqxmd.com/read/28647581/development-of-field-applicable-tests-for-rapid-and-sensitive-detection-of-candidatus-phytoplasma-oryzae
#8
Lillian Wambua, Bernd Schneider, Allan Okwaro, Joseph Odhiambo Wanga, Olive Imali, Peninah Nduku Wambua, Lavender Agutu, Cassandra Olds, Chris Stephen Jones, Daniel Masiga, Charles Midega, Zeyaur Khan, Joerg Jores, Anne Fischer
Napier grass Stunt Disease (NSD) is a severe disease of Napier grass (Pennisetum purpureum) in Eastern Africa, caused by the leafhopper-transmitted bacterium Candidatus Phytoplasma oryzae. The pathogen severely impairs the growth of Napier grass, the major fodder for dairy cattle in Eastern Africa. NSD is associated with biomass losses of up to 70% of infected plants. Diagnosis of NSD is done by nested PCR targeting the phytoplasma DNA, which is difficult to perform in developing countries with little infrastructure...
October 2017: Molecular and Cellular Probes
https://www.readbyqxmd.com/read/28627450/impact-of-fixation-artifacts-and-threshold-selection-on-high-resolution-melting-analysis-for-kras-mutation-screening
#9
Wendy Pérez-Báez, Ethel A García-Latorre, Héctor Aquiles Maldonado-Martínez, Iris Coronado-Martínez, Leonardo Flores-García, Lucía Taja-Chayeb
INTRODUCTION: Treatment in metastatic colorectal cancer (mCRC) has expanded with monoclonal antibodies targeting epidermal growth factor receptor, but is restricted to patients with a wild-type (WT) KRAS mutational status. The most sensitive assays for KRAS mutation detection in formalin-fixed paraffin embedded (FFPE) tissues are based on real-time PCR. Among them, high resolution melting analysis (HRMA), is a simple, fast, highly sensitive, specific and cost-effective method, proposed as adjunct for KRAS mutation detection...
October 2017: Molecular and Cellular Probes
https://www.readbyqxmd.com/read/28627449/microrna-214-regulates-immunity-related-genes-in-bovine-mammary-epithelial-cells-by-targeting-nfatc3-and-traf3
#10
Ning Song, Xingping Wang, Linsheng Gui, Sayed Haidar Abbas Raza, Zhuoma Luoreng, Linsen Zan
In human, microRNA-214 (miR-214) plays crucial roles in mechanisms of immunity. However, the potential importance of miR-214 in immune mechanisms in dairy cows has not been investigated. In this study, we assessed potential immunity-related functions of miR-214 in human 293A cells and in bovine mammary epithelial cells (BMECs). We found that NFATc3 and TRAF3 could be targeted by miR-214 in both 293A cells and BMECs. We also found that miR-214 indirectly inhibited the expression of MAP3K14, TBK1 and inflammatory cytokines IL-6 and IL-1β...
October 2017: Molecular and Cellular Probes
https://www.readbyqxmd.com/read/28587995/duffy-blood-group-system-new-genotyping-method-and-distribution-in-a-brazilian-extra-amazonian-population
#11
Marina Lobato Martins, Adão Rogerio da Silva, Hadassa Campos Santos, Michelle Teodoro Alves, Luciana Cayres Schmidt, Stela Brener Vertchenko, Luci Maria SantAna Dusse, Maria Clara Fernandes da Silva Malta
Duffy blood group system is of interest in several fields of science including transfusion medicine, immunology and malariology. Although some methods have been developed for Duffy polymorphism genotyping, not all of them have been sufficiently described and validated, and all present limitations. At the same time, the frequency of Duffy alleles and antigens in some densely populated regions of the world are still missing. In this study we present new tests for genotyping the major alleles of the Duffy blood system and describe Duffy alleles and antigens in blood donors and transfusion-dependent patients in Minas Gerais, Brazil...
October 2017: Molecular and Cellular Probes
https://www.readbyqxmd.com/read/28578011/homology-modeling-and-in-silico-prediction-of-ulcerative-colitis-associated-polymorphisms-of-nod1
#12
Ishani Majumdar, Isha Nagpal, Jaishree Paul
Cytosolic pattern recognition receptors play key roles in innate immune response. Nucleotide binding and oligomerisation domain containing protein 1 (NOD1) belonging to the Nod-like receptor C (NLRC) sub-family of Nod-like receptors (NLRs) is important for detection and clearance of intra-cellular Gram negative bacteria. NOD1 is involved in activation of pro-inflammatory pathways. Limited structural data is available for NOD1. Using different templates for each domain of NOD1, we determined the full-length homology model of NOD1...
October 2017: Molecular and Cellular Probes
https://www.readbyqxmd.com/read/28558918/general-phytoplasma-detection-by-a-q-pcr-method-using-mycoplasma-primers
#13
Eleonora Satta, Irene M Nanni, Nicoletta Contaldo, Marina Collina, José B Poveda, Ana S Ramírez, Assunta Bertaccini
Phytoplasmas and mycoplasmas are bacteria belonging to the class Mollicutes. In this study, a fine tuning of quantitative polymerase chain reaction (qPCR) with a universal mycoplasma primer pair (GPO3F/MGSO) targeting the 16S rRNA gene was carried out on phytoplasmas. The dissociation curves of DNAs from Catharanthus roseus phytoplasma-infected micropropagated shoots and from phytoplasma field-infected plant samples showed a single peak at 82.5 °C (±0.5) specifically detecting phytoplasmas belonging to several ribosomal groups...
October 2017: Molecular and Cellular Probes
https://www.readbyqxmd.com/read/28734841/potential-involvement-of-mi-rna-574-3p-in-progression-of-prostate-cancer-a-bioinformatic-study
#14
Naeem M Ashraf, Komal Imran, David W Kastner, Khadija Ikram, Ayesha Mushtaq, Aadil Hussain, Nadia Zeeshan
Aberrant gene expression is a hallmark of prostate cancer (PCa), the second deadliest disease affecting males worldwide. Dysregulation of miRNA has been associated with the progression of PCa and in recent studies, miRNA 574-3p was found to be upregulated in cancerous prostate tissue. In this study, we characterize the effects of upregulated miRNA 574-3p on gene expression in the tumor microenvironment through different bioinformatic tools such as Diana-Tools, the KEGG Pathway Database, and the Reactome Database...
July 19, 2017: Molecular and Cellular Probes
https://www.readbyqxmd.com/read/28698006/identification-of-tradd-as-a-potential-biomarker-in-human-uterine-leiomyoma-through-itraq-based-proteomic-profiling
#15
Yong Liu, Dan Lu, Jie Sheng, Lijing Luo, Weiyuan Zhang
Recurrent and refractory leiomyoma of uterus is one of the most common diseases in women of reproductive age. Despite its benign nature, uterine leiomyoma has presented an extremely deleterious impact on public health. The etiology of uterine leiomyoma remains unclear and clinical management remains suboptimal, leaving radical hysterectomy the only effective approach. Delineating the molecular mechanism underlying the leiomyoma initiation and progression remains an unmet clinical need. To screen proteins that were differentially expressed in uterine leiomyoma versus normal myometrium, we examined proteomic profile by isobaric tag for relative and absolute quantitation (iTRAQ) labeling coupled with liquid chromatography - tandem mass spectrometry (LC-MS/MS)...
July 8, 2017: Molecular and Cellular Probes
https://www.readbyqxmd.com/read/28668278/synthetic-scale-up-of-a-novel-fluorescent-probe-and-its-biological-evaluation-for-surface-detection-of-staphylococcus-aureus
#16
Luke Bywaters, Lauren Mulcahy-Ryan, Mark Fielder, Alex Sinclair, Adam Le Gresley
This paper reports on the LGX fluorometric test for enzymatic MRSA/MSSA detection. It highlights the reasons rhodamines have been overlooked and also strategies to improve the synthesis of rhodamine-peptide conjugates. Evaluation of the LGX test for detection of MRSA/MSSA on surfaces is undertaken in the presence of potentially confounding E. coli and S. epidermidis for the first time.
June 28, 2017: Molecular and Cellular Probes
https://www.readbyqxmd.com/read/28666619/development-of-a-multiplex-pcr-for-identification-of-%C3%AE-hemolytic-streptococci-relevant-to-human-infections-and-serotype-distribution-of-invasive-streptococcus-agalactiae-in-thailand
#17
Anusak Kerdsin, Rujirat Hatrongjit, Shigeyuki Hamada, Yukihiro Akeda, Marcelo Gottschalk
A multiplex polymerase chain reaction (mPCR) was developed for simultaneous detection (single reaction) of genes specific to five frequent clinically relevant β-hemolytic streptococcal species: Streptococcus pyogenes (Spy1258), Streptococcus agalactiae (cfb and cpn60), Streptococcus dysgalactiae subsp. equisimilis (16S-23S intergenic spacer) , S. equi subsp. zooepidemicus (esaA and sorD), and Streptococcus anginosus group (moaC). No cross-reaction was observed with other bacterial species. This test was validated and successfully used with 725 clinical isolates involved in pathological conditions in Thailand and collected between March 2014 and December 2015...
June 27, 2017: Molecular and Cellular Probes
https://www.readbyqxmd.com/read/28576288/editor-note
#18
EDITORIAL
(no author information available yet)
No abstract text is available yet for this article.
June 2017: Molecular and Cellular Probes
https://www.readbyqxmd.com/read/28411089/evidence-for-polymicrobial-communities-in-explanted-vascular-filters-and-atheroma-debris
#19
Jeremy E Ellis, Richard Heuser, Dara S Missan, Delyn Martinez, Avory Heningburg, Matthew Shabilla, Renata Schwartz, Stephen Fry
RATIONALE: Microbial communities have been implicated in a variety of disease processes and have been intermittently observed in arterial disease; however, no comprehensive unbiased community analysis has been performed. We hypothesize that complex microbial communities may be involved in chronic vascular diseases as well and may be effectively characterized by molecular assays. OBJECTIVE: The main objective is to survey vascular debris, atheroma, and vascular filters for polymicrobial communities consisting of prokaryotic and eukaryotic microbes, specifically eukaryotic microbes...
June 2017: Molecular and Cellular Probes
https://www.readbyqxmd.com/read/28342800/development-of-an-isothermal-recombinase-polymerase-amplification-assay-for-rapid-detection-of-pseudorabies-virus
#20
Yang Yang, Xiaodong Qin, Wei Zhang, Zhiyong Li, Shuaijun Zhang, Yanmin Li, Zhidong Zhang
Recombinase polymerase amplification assays using real-time fluorescent detection (real-time RPA assay) and lateral flow dipstick (RPA LFD assay) were developed targeting the gD gene of pseudorabies virus (PRV). Both assays were performed at 39 °C within 20 min. The sensitivity of the real-time RPA assay and the RPA LFD assay was 100 copies per reaction and 160 copies per reaction, respectively. Both assays did not detect DNAs from other virus or PRV negative samples. Therefore, the developed RPA assays provide a rapid, simple, sensitive and specific alternative tool for detection of PRV...
June 2017: Molecular and Cellular Probes
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