Search results for "rt-PCR"
showing 10 items of 44 documents
Molecular characterization of cucumber mosaic virus isolates infecting ornamental species cultivated in the botanical garden of Bologna university.
2006
During an epidemiological survey carried out in the Botanical Garden at Bologna University, Cucumber mosaic virus (CMV) has been detected by applying PAS-ELISA technique infecting some ornamental species exhibiting a severe symptomatology on the leaves. Datura innoxia Miller. showed mosaic and leaf-curling; Globularia nudicaulis L. produced narrowed leaves with a yellow mosaic and/or variegation; Eupatorium cannabinum L. showed a systemic chlorotic and/or yellow mosaic and stunting. Reverse transcription-polymerase chain reaction (RT-PCR) and single strand conformation polymorphism (SSCP) were employed to characterise these CMV isolates. Total RNA was extracted from symptomatic leaf samples…
Proposal of a Citrus translational genomic approach for early and infield detection of Flavescence dorée in Vitis
2014
Flavescence dore´e (FD) is one of the most widely known grapevine yellows disease and one of the most unabated worldwide in the viticulture sector. In this paper, we outline a strategy for developing an integrated system of technologies to enable rapid, early disease FD detection and diagnosis. We propose the deployment of a newly developed sensor device, the differential mobility spectrometer (DMS), which has shown positive results with a similar vector-borne disease in Citrus. We have previously demonstrated that the gas chromatograph DMS (GC/DMS) can distinguish various citrus diseases, and the system may also allow detection of volatile organic compound (VOC) signals from a tree of othe…
Detection and identification Fabavirus species by one-step RT-PCR and multiplex RT-PCR
2014
The genus Fabavirus of the family Secoviridae comprises a group of poorly characterized viruses. To date, only five species have been described: Broad bean wilt virus 1 (BBWV-1), Broad bean wilt virus 2 (BBWV-2), Lamium mild mosaic virus (LMMV), Gentian mosaic virus (GeMV) and Cucurbit mild mosaic virus (CuMMV). The development is described of two RT-PCR procedures for the detection and identification of Fabavirus species: a one-step RT-PCR using a single pair of conserved primers for the detection of all fabaviruses, and a one-step multiplex RT-PCR using species-specific primers for the simultaneous detection and identification of the above-mentioned species of the genus Fabavirus. These m…
Implementation of Sample Pooling Procedure Using a Rapid SARS-CoV-2 Diagnostic Real-Time PCR Test Performed Prior to Hospital Admission of People wit…
2021
Reliability, accuracy, and timeliness of diagnostic testing for SARS-CoV-2 infection have allowed adequate public health management of the disease, thus notably helping the timely mapping of viral spread within the community. Furthermore, the most vulnerable populations, such as people with intellectual disability and dementia, represent a high-risk group across multiple dimensions, including a higher prevalence of pre-existing conditions, lower health maintenance, and a propensity for rapid community spread. This led to an urgent need for reliable in-house rapid testing to be performed prior to hospital admission. In the present study, we describe a pooling procedure in which oropharyngeal…
CHARACTERIZATION OF A STRAIN OF PEPINO MOSAIC VIRUS FOUND IN SICILY
2006
During winter 2005, an unusual virus-like yellow leaf disorder associated to interveinal leaf yellowing and marbling on fruits, was observed in some greenhouse tomato crops in Sicily, exactly in province of Ragusa. Two leaves from 250 symptomatic tomato plants were serologically tested by DAS-ELISA technique using the sera to Tomato spotted wilt virus (TSWV), Impatiens necrotic spot virus (INSV), Tobacco mosaic virus (TMV), Cucumber mosaic virus (CMV) and Pepino mosaic virus (PepMV). Of the 250 samples tested, 215 were positive to PepMV presence. This virus was mechanically transmitted to Cucumis sativus “Cubit”, Datura metel, Nicotiana benthamiana and tomato “Rio Grande”. The host range of…
Mboat7 down-regulation by hyper-insulinemia induces fat accumulation in hepatocytes.
2020
Background: Naturally occurring variation in Membrane-bound O-acyltransferase domain-containing 7 (MBOAT7), encoding for an enzyme involved in phosphatidylinositol acyl-chain remodelling, has been associated with fatty liver and hepatic disorders. Here, we examined the relationship between hepatic Mboat7 down-regulation and fat accumulation. Methods: Hepatic MBOAT7 expression was surveyed in 119 obese individuals and in experimental models. MBOAT7 was acutely silenced by antisense oligonucleotides in C57Bl/6 mice, and by CRISPR/Cas9 in HepG2 hepatocytes. Findings: In obese individuals, hepatic MBOAT7 mRNA decreased from normal liver to steatohepatitis, independently of diabetes, inflammatio…
Polymerase Chain Reaction (PCR)
2008
Comparison of the BD Directigen Flu A+B Kit and the Abbott TestPack RSV with a multiplex RT-PCR ELISA for rapid detection of influenza viruses and re…
2005
ABSTRACTThe Directigen Flu A+B enzyme immunoassay and the Abbott TestPack RSV enzyme immunoassay were each compared with a multiplex RT-PCR ELISA by testing 635 nasopharyngeal aspirates collected from children aged < 16 years who had been hospitalised with acute respiratory tract infection during the epidemic season 2002–2003. In this study, the sensitivity of the Directigen Flu A+B assay was unacceptably low (29.3% and 10.0%, respectively) for the detection of influenza A and B viruses. The sensitivity of the Abbott TestPack RSV assay (77.4%) was acceptable and in agreement with the multiplex RT-PCR ELISA.
CD40 activation in human pancreatic islets and ductal cells.
2008
Aims/hypothesis: CD40 expression on non-haematopoietic cells is linked to inflammation. We previously reported that CD40 is expressed on isolated human and non-human primate islets and its activation results in secretion of IL-8, macrophage inflammatory protein 1-beta (MIP-1β) and monocyte chemoattractant protein-1 (MCP-1) through nuclear factor-κB and extracellularly regulated kinases 1/2 pathways. The objective of this study was to identify the pattern of gene expression, and to study viability and functionality affected by CD40-CD40 ligand (CD40L) interaction in human islets. Furthermore, we have studied the CD40-mediated cytokine/chemokine profile in pancreatic ductal cells, as they are…
Expression in Streptomyces lividans of Nonomuraea genes cloned in an artificial chromosome
2004
A bacterial artificial chromosomal library of Nonomuraea sp. ATCC39727 was constructed using Escherichia coli-Streptomyces artificial chromosome (ESAC) and screened for the presence of dbv genes known to be involved in the biosynthesis of the glycopeptide A40926. dbv genes were cloned as two large, partially overlapping, fragments and transferred into the host Streptomyces lividans, thus generating strains S. lividansColon, two colonsNmESAC50 and S. lividansColon, two colonsNmESAC57. The heterologous expression of Nonomuraea genes in S. lividans was successfully demonstrated by using combined RT-PCR and proteomic approaches. MALDI-TOF analysis revealed that a Nonomuraea ABC transporter is e…