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Molecular and Cellular Probes

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https://www.readbyqxmd.com/read/28526589/a-rapid-salmonella-detection-method-involving-thermophilic-helicase-dependent-amplification-and-a-lateral-flow-assay
#1
Xin-Jun Du, Tian-Jiao Zhou, Ping Li, Shuo Wang
Salmonella is a major foodborne pathogen that is widespread in the environment and can cause serious human and animal disease. Since conventional culture methods to detect Salmonella are time-consuming and laborious, rapid and accurate techniques to detect this pathogen are critically important for food safety and diagnosing foodborne illness. In this study, we developed a rapid, simple and portable Salmonella detection strategy that combines thermophilic helicase-dependent amplification (tHDA) with a lateral flow assay to provide a detection result based on visual signals within 90 min...
May 16, 2017: Molecular and Cellular Probes
https://www.readbyqxmd.com/read/28502787/no-association-detected-between-seven-common-variants-in-the-cdkal1-gene-and-gestational-glycemic-traits
#2
Hongfang Ju, Rong Lin, Ziyu Yuan, Yuantian Sun, Liangliang Zeng, Zhenyu Su, Li Jin
Elevated plasma glucose levels in pregnancy increase adverse pregnancy outcomes. Cyclin-dependent kinase 5 regulatory subunit-associated protein 1-like 1 (CDKAL1) has been shown to be involved in insulin secretion in pancreatic β cells. In this study, we investigated the impact of genetic variants in CDKAL1 on plasma glucose, insulin values, β cell function and insulin resistance in the fasted state as well as plasma glucose 1 h after the consumption of a 50-g oral glucose load between 24 and 28 weeks of pregnancy among 929 unrelated pregnant Han Chinese women...
May 11, 2017: Molecular and Cellular Probes
https://www.readbyqxmd.com/read/28499542/development-of-a-taqman-based-real-time-pcr-assay-for-the-rapid-and-specific-detection-of-novel-duck-origin-goose-parvovirus
#3
Jianchang Wang, Jinfeng Wang, Yuan Cui, Huizhu Nan, Wanzhe Yuan
A real-time PCR assay was developed for specific detection of novel duck-origin goose parvovirus (N-GPV), the etiological agent of duck beak atrophy and dwarfism syndrome (BADS). The detection limit of the assay was 10(2) copies. The assay was useful in the prevention and control of BADS.
May 9, 2017: Molecular and Cellular Probes
https://www.readbyqxmd.com/read/28499541/sequence-analysis-of-haemagglutinin-and-neuraminidase-of-h1n1-strain-from-a-patient-coinfected-with-mycobacterium-tuberculosis
#4
Ahmed N Alghamdi, Mohammad E Mahfouz, Fahd A Hamdi, Daifullah Al Aboud, Tawfiq Z Al-Laylah, Mohammed I Alotaibi, Khalid W A Al-Thomali, Ahmed S Abdel-Moneim
The 2009 H1N1 pandemic (H1N1pdm09) was associated with a considerable influenza-related morbidity and mortality. Among the complications, Mycobacterial tuberculosis was recorded as a coinfection with influenza in rare cases. The full-length sequences of the viral haemagglutinin and neuraminidase of H1N1pdm09 influenza A virus were analyzed from a recently infected patient. The patient was chronically infected with Mycobacterium tuberculosis. Molecular modelling and in-silico docking of the virus, and other selected strains with the drug oseltamivir were conducted and compared...
May 9, 2017: Molecular and Cellular Probes
https://www.readbyqxmd.com/read/28461258/thiolated-aunp-probes-and-multiplex-pcr-for-molecular-detection-of-staphylococcus-epidermidis
#5
Mahsa Osmani Bojd, Hossein Kamaladini, Fatemeh Haddadi, Akbar Vaseghi
The emergence of nanotechnology in biology helps to apply the gold nanoparticle probes for fast and accurate identification of pathogens compared to the time-consuming and non-precise phenotypic methods. In this study, two molecular methods have been established for the accurate identification of staphylococcus epidermidis from other coagulase-negative staphylococci. Multiplex PCR was performed using designed primers for Gmk2 and pta housekeeping genes, and SESB specific gene of S. epidermidis. Colorimetric detection by gold nanoparticle probes was carried out using two 20-base thiolated probes designed based on the sequence of pta housekeeping gene of S...
April 28, 2017: Molecular and Cellular Probes
https://www.readbyqxmd.com/read/28457983/assessment-of-ns1-gene-specific-real-time-quantitative-taqman-pcr-for-the-detection-of-human-bocavirus-in-respiratory-samples
#6
Marco Ligozzi, Erica Diani, Federica Lissandrini, Rosanna Mainardi, Davide Gibellini
Human Bocaviruses (HBoV) were associated with respiratory diseases. Here, we assessed a TaqMan(®)-based PCR for the detection of all four HBoV subtype infections with a sensitivity up to 15 copies/reaction. To evaluate this assay on clinical samples, 178 nasopharyngeal aspirate specimens from pediatric cases were analyzed and HBoV genome was detected in 13 out of 178 patients with a viral load range between 1.6 × 10(3) and 9.4 × 10(7) copies/ml. These results indicated that this method could be used as an alternative technique for the diagnosis of HBoV infection...
April 27, 2017: Molecular and Cellular Probes
https://www.readbyqxmd.com/read/28455235/computational-prediction-and-analysis-of-deleterious-cancer-associated-missense-mutations-in-dync1h1
#7
Ceren Sucularli, Melda Arslantas
Dynein, cytoplasmic 1, heavy chain 1 (DYNC1H1) gene encodes a subunit of human cytoplasmic dynein complex, which has several crucial functions in the cell, such as intracellular transport of DNA damage proteins and mitotic spindle positioning. Recent studies reported the altered expression of DYNC1H1 in different cancers and DYNC1H1 was suggested to be potential biomarker in colorectal cancers. Previously, DYNC1H1 mutations have been associated with neurodegenerative diseases, however mutations of DYNC1H1 have not been fully investigated in cancers except for different types of pancreatic cancers...
April 25, 2017: Molecular and Cellular Probes
https://www.readbyqxmd.com/read/28433811/rapid-and-sensitive-detection-of-salmonid-alphavirus-using-taqman-real-time-pcr
#8
Wen Shi, Aochen Song, Shuai Gao, Yuting Wang, Lijie Tang, Yigang Xu, Tong Ren, Yijing Li, Min Liu
Salmonid alphavirus (SAV) infection has led to the spread of salmon pancreas disease (PD) and sleeping disease (SD) to salmonids in several countries in Europe, resulting in tremendous economic losses to the fish farming industry. Recently, with increases in the fish import trade, many countries in which SAV has been unreported, such as China, may be seriously threatened by these diseases. It is therefore necessary to develop efficient detection methods for the prevention and diagnosis of SAV infection. In this study, a rapid and sensitive TaqMan real-time PCR method was established and assessed for this purpose...
April 19, 2017: Molecular and Cellular Probes
https://www.readbyqxmd.com/read/28411089/evidence-for-polymicrobial-communities-in-explanted-vascular-filters-and-atheroma-debris
#9
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...
April 11, 2017: Molecular and Cellular Probes
https://www.readbyqxmd.com/read/28400333/umbilical-cord-mesenchymal-stem-cells-are-able-to-undergo-differentiation-into-functional-islet-like-cells-in-type-2-diabetic-tree-shrews
#10
Xing-Hua Pan, Xia Huang, Guang-Ping Ruan, Rong-Qing Pang, Qiang Chen, Jin-Xiang Wang, Jie He, Jing Zhao, Xue-Min Cai, Na Zhao, Yi Chen, Xiang-Qing Zhu
Islet transplantation is arguably one of the most promising strategies to treat patients suffering with diabetes mellitus. However, a combination of a lack of donors and chronic immune rejection limit clinical applications. Here, we evaluated the efficacy of cell therapy using islet-like cells differentiated from umbilical cord mesenchymal stem cells (UC-MSCs) of tree shrews for the treatment of type 2 diabetes. Enhanced green fluorescent protein (eGFP) labeled UC-MSCs were directly injected into type 2 diabetic tree shrews, where UC-MSC differentiated into functional islet-like cells and alleviated disease severity, as evidenced by improved biochemical features and reduced concentrations of inflammatory cytokines...
April 8, 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
#11
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...
March 22, 2017: Molecular and Cellular Probes
https://www.readbyqxmd.com/read/28336361/a-novel-multiplex-pcr-assay-for-simultaneous-detection-of-nine-clinically-significant-bacterial-pathogens-associated-with-bovine-mastitis
#12
Aqeela Ashraf, Muhammad Imran, Tahir Yaqub, Muhammad Tayyab, Wasim Shehzad, Peter C Thomson
For rapid and simultaneous detection of nine bovine mastitic pathogens, a sensitive and specific multiplex PCR assay was developed. The assay was standardized using reference strains and validated on mastitic milk cultures which were identified to species level based on 16S rRNA sequencing. Multiplex PCR assay also efficiently detected the target bacterial strains directly from milk. Detection limit of the assay was up to 50 pg for DNA isolated from pure cultures and 10(4) CFU/ml for spiked milk samples. As estimated by latent class analysis, the assay was sensitive up to 88% and specific up to 98% for targeted mastitic pathogens, compared with the bacterial culture method and the 16S rRNA sequence analysis...
March 20, 2017: Molecular and Cellular Probes
https://www.readbyqxmd.com/read/28302392/osteopontin-regulates-the-proliferation-of-rat-aortic-smooth-muscle-cells-in-response-to-gingipains-treatment
#13
Chong Cao, Xin Luo, Xiaowei Ji, Yao Wang, Yuan Zhang, Pengtao Zhang, Liangjun Zhong
OBJECTIVE: The present study aimed to explore the possible effects of osteopontin (OPN) in the proliferation of rat aortic smooth muscle cells (RASMCs) stimulated by gingipains. METHODS: The proliferation of RASMCs in response to active gingipains treatment was evaluated by CCK-8 assay. OPN siRNA was designed, constructed and transfected into RASMCs at different concentrations. The cell cycle of RASMCs was analyzed by flow cytometry. OPN, α-SMA and calponin expression were examined by real-time PCR and western blot analysis...
March 13, 2017: Molecular and Cellular Probes
https://www.readbyqxmd.com/read/28267624/a-taqman-based-qrt-pcr-assay-for-senecavirus-a-detection-in-tissue-samples-of-neonatal-piglets
#14
Alais Maria Dall Agnol, Rodrigo Alejandro Arellano Otonel, Raquel Arruda Leme, Amauri Alcindo Alfieri, Alice Fernandes Alfieri
This study describes a sensitive (1.3 × 10(1) genomic copies/μL) and specific TaqMan-based qRT-PCR assay able to detect and quantify SVA RNA in porcine biological samples. The technique represents an efficient tool for the virus diagnosis and assessment of SVA load in tissues of infected animals and for epidemiological studies.
March 4, 2017: Molecular and Cellular Probes
https://www.readbyqxmd.com/read/28263784/putative-tmprss3-gjb2-digenic-inheritance-of-hearing-loss-detected-by-targeted-resequencing
#15
Maria Pia Leone, Pietro Palumbo, Rocco Ortore, Stefano Castellana, Orazio Palumbo, Salvatore Melchionda, Teresa Palladino, Raffaella Stallone, Tommaso Mazza, Roberto Cocchi, Massimo Carella
The paper describes a putative digenic form of deafness in two siblings affected by non-syndromic hereditary hearing loss, detected by a Targeted resequencing approach. Given that a previous paper suggested TMPRSS3 and GJB2 genes as responsible for a digenic form of hearing loss, our data support and reinforce this hypothesis.
March 3, 2017: Molecular and Cellular Probes
https://www.readbyqxmd.com/read/28254505/mt-pcr-panel-detection-of-canine-parvovirus-cpv-2-vaccine-and-wild-type-cpv-2-can-be-difficult-to-differentiate-in-canine-diagnostic-fecal-samples
#16
Maira N Meggiolaro, Anna Ly, Benjamin Rysnik-Steck, Carolina Silva, Joshua Zhang, Damien P Higgins, Gary Muscatello, Jacqueline M Norris, Mark Krockenberger, Jan Šlapeta
Canine parvovirus (CPV-2) remains an important cause of devastating enteritis in young dogs. It can be successfully prevented with live attenuated CPV-2 vaccines when given at the appropriate age and in the absence of maternal antibody interference. Rapid diagnosis of parvoviral enteritis in young dogs is essential to ensuring suitable barrier nursing protocols within veterinary hospitals. The current diagnostic trend is to use multiplexed PCR panels to detect an array of pathogens commonly responsible for diarrhea in dogs...
February 27, 2017: Molecular and Cellular Probes
https://www.readbyqxmd.com/read/28232024/selection-and-validation-of-candidate-reference-genes-for-quantitative-real-time-pcr-studies-in-the-shrimp-penaeus-vannamei-under-viral-infection
#17
Adán Valenzuela-Castillo, Fernando Mendoza-Cano, Tania Enríquez-Espinosa, José Manuel Grijalva-Chon, Arturo Sánchez-Paz
The decapod Penstyldensovirus 1 (PstDV-1) represents one of the most serious threats for penaeid shrimp farming. Studies aimed at defining relevant molecular effects of this virus over its host are imperative in the attempt to increase our understanding of its pathogenesis. Unfortunately, few studies have focused on the definition of the expression profile of reference genes in shrimp challenged with a pathogen. As a result, there are no studies on the selection of reference genes for the normalization of target gene expression changes yielding reliable data of the effects following PstDV-1 infection in shrimp...
February 20, 2017: Molecular and Cellular Probes
https://www.readbyqxmd.com/read/28223170/development-of-taqman-rt-qpcr-for-the-detection-of-type-a-human-respiratory-syncytial-virus
#18
Ahmed S Abdel-Moneim, Gaber M Shehab, Adnan A Alsulaimani, Mater I R Al-Malky
The human respiratory syncytial virus is a common respiratory pathogen in children. Improved diagnosis of the virus is dependent on the development of tools for the rapid detection and estimation of the viral loads. In the current study, RT-qPCR using TaqMan hydrolysis probe based on the F gene detection was developed to identify and quantify hRSV in clinical samples. The assay was validated by comparing the results with a commercially available RT-qPCR kit. The newly developed assay was sensitive in detecting hRSV positive samples (59/126) which were equivalent to those detected by the commercial kit (57/126) with a detection limit of 1 × 10(2) copies/mL...
February 20, 2017: Molecular and Cellular Probes
https://www.readbyqxmd.com/read/28192165/copy-number-variation-of-housekeeping-gene-rpl13a-in-rat-strains-selected-for-nervous-system-excitability
#19
Ruslan Kalendar, Alexander Belyayev, Tatiana Zachepilo, Alexander Vaido, Dmitry Maidanyuk, Alan H Schulman, Natalia Dyuzhikova
We evaluated copy number variation (CNV) for four genes in rat strains differing in nervous system excitability. rpl13a copy number is significantly reduced in hippocampus and bone marrow in rats with a high excitability threshold and stress. The observed phenomenon may be associated with a role for rpl13a in lipid metabolism.
February 10, 2017: Molecular and Cellular Probes
https://www.readbyqxmd.com/read/28189797/rapid-detection-of-enterovirus-and-coxsackievirus-a10-by-a-taqman-based-duplex-one-step-real-time-rt-pcr-assay
#20
Jingfang Chen, Rusheng Zhang, Xinhua Ou, Dong Yao, Zheng Huang, Linzhi Li, Biancheng Sun
A TaqMan based duplex one-step real time RT-PCR (rRT-PCR) assay was developed for the rapid detection of Coxsackievirus A10 (CV-A10) and other enterovirus (EVs) in clinical samples. The assay was fully evaluated and found to be specific and sensitive. When applied in 115 clinical samples, a 100% diagnostic sensitivity in CV-A10 detection and 97.4% diagnostic sensitivity in other EVs were found.
February 8, 2017: Molecular and Cellular Probes
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