Rapid detection of Newcastle disease virus replication in embryonated chicken eggs using quantitative real time polymerase chain reaction

Gopinath, V.P.; Raj, Gopal Dhinakar; Amri, R.; Kumanan, K.; Elankumaran, Subbiah

doi:10.1016/j.jviromet.2010.10.007

Cited by 13 publications

(3 citation statements)

References 10 publications

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“…The slope values were calculated using serial dilutions of cDNA and the respective Ct values for each dilution and PCR efficiency (E = 10 21 /slope) was determined. Results were expressed as 40-Ct values [54,55]. Changes in cytokine expression were expressed as fold change (2 2DDCt ) over the respective basal levels of mockinduced PBMCs after normalizing for the endogenous control gene and using the corrected Ct.…”

Section: Cytokine Transcripts After Stimulation With Tlr Agonists or mentioning

confidence: 99%

Toll-Like Receptor Responses to Peste des petits ruminants Virus in Goats and Water Buffalo

et al. 2014

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Ovine rinderpest or goat plague is an economically important and contagious viral disease of sheep and goats, caused by the Peste des petits ruminants virus (PPRV). Differences in susceptibility to goat plague among different breeds and water buffalo exist. The host innate immune system discriminates between pathogen associated molecular patterns and self antigens through surveillance receptors known as Toll like receptors (TLR). We investigated the role of TLR and cytokines in differential susceptibility of goat breeds and water buffalo to PPRV. We examined the replication of PPRV in peripheral blood mononuclear cells (PBMC) of Indian domestic goats and water buffalo and demonstrated that the levels of TLR3 and TLR7 and downstream signalling molecules correlation with susceptibility vs resistance. Naturally susceptible goat breeds, Barbari and Tellichery, had dampened innate immune responses to PPRV and increased viral loads with lower basal expression levels of TLR 3/7. Upon stimulation of PBMC with synthetic TLR3 and TLR7 agonists or PPRV, the levels of proinflammatory cytokines were found to be significantly higher while immunosuppressive interleukin (IL) 10 levels were lower in PPRV resistant Kanni and Salem Black breeds and water buffalo at transcriptional level, correlating with reduced viralloads in infected PBMC. Water buffalo produced higher levels of interferon (IFN) α in comparison with goats at transcriptional and translational levels. Pre-treatment of Vero cells with human IFNα resulted in reduction of PPRV replication, confirming the role of IFNα in limiting PPRV replication. Treatment with IRS66, a TLR7 antagonist, resulted in the reduction of IFNα levels, with increased PPRV replication confirming the role of TLR7. Single nucleotide polymorphism analysis of TLR7 of these goat breeds did not show any marked nucleotide differences that might account for susceptibility vs resistance to PPRV. Analyzing other host genetic factors might provide further insights on susceptibility to PPRV and genetic polymorphisms in the host.

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Section: Cytokine Transcripts After Stimulation With Tlr Agonists or mentioning

confidence: 99%

Toll-Like Receptor Responses to Peste des petits ruminants Virus in Goats and Water Buffalo

et al. 2014

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“…The qRT-PCR of all samples was performed according to the previously reported protocols [33, 34]. Briefly, RNA was extracted using the RNAqueous-Micro Kit (Ambion USA) according to the manufacturer's instructions.…”

Section: Methodsmentioning

confidence: 99%

Viability Reduction andRac1Gene Downregulation of HeterogeneousEx-VivoGlioma Acute Slice Infected by the Oncolytic Newcastle Disease Virus Strain V4UPM

Mustafa

Shamsuddin

Ideris

et al. 2013

BioMed Research International

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Oncolytic viruses have been extensively evaluated for anticancer therapy because this virus preferentially infects cancer cells without interfering with normal cells. Newcastle Disease Virus (NDV) is an avian virus and one of the intensively studied oncolytic viruses affecting many types of cancer including glioma. Nevertheless, the capability of NDV infection on heterogeneous glioma tissue in a cerebrospinal fluid atmosphere has never been reported. Recently, Rac1 is reported to be required for efficient NDV replication in human cancer cells and established a link between tumourigenesis and sensitivity to NDV. Rac1 is a member of the Rho GTPases involved in the regulation of the cell migration and cell-cycle progression. Rac1 knockdown leads to significant inhibition of viral replication. In this work, we demonstrated that NDV treatment led to significant reduction of tumour tissue viability of freshly isolated heterogeneous human brain tumour slice, known as an ex vivo glioma acute slice (EGAS). Analysis of gene expression indicated that reduced tissue viability was associated with downregulation of Rac1. However, the viability reduction was not persistent. We conclude that NDV treatment induced EGAS viability suppression, but subsequent downregulation of Rac1 gene may reduce the NDV replication and lead to regrowth of EGAS tissue.

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“…Besides high sensitivity and readout formats suitable for non-research laboratories, an increasing demand for real-time assays for quantification of clinically relevant analytes has been put forward. Whereas such assays have been successfully developed for nucleic acid targets in the form of qPCR and qRT-PCR assays, [184][185][186][187][188] real-time assays for analysis of proteins or other non-nucleotide biomolecules have been more difficult to achieve. 189 RCA combined with nucleotide biosensors targeting vast varieties of biomolecules may, however, enable real-time protein/small molecule detection.…”

Section: Readout Formats For Detection Of Rcpsmentioning

confidence: 99%

Strategies for highly sensitive biomarker detection by Rolling Circle Amplification of signals from nucleic acid composed sensors

et al. 2011

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The use of nucleic acids as components in highly sensitive biosensors has attracted increasing interest during recent years, not least due to the ease by which nucleic acids can be synthesized, manipulated and signal-amplified. To date, several enzymatic reactions, including Polymerase Chain Reaction, Rolling Circle- and Strand Displacement Amplification for signal enhancement of nucleic acid biosensors, have been presented. Of these, the isothermal Rolling Circle Amplification, in which a small single-stranded DNA circle is replicated to generate long tandem repeat products, presents the advantage of allowing single molecule detection and easy quantification as well as of posing little requirement to equipment and personnel training. Rolling Circle Amplification has been used to enhance signals of nucleic acid based sensors specific for a large variety of important biomarkers, including nucleotide sequences, small molecules, proteins and enzyme activities, allowing detection of biomarkers even at the aM concentration level. Moreover, Rolling Circle Products have been adapted to a broad variety of low technological and cost efficient readout formats making the technique appealing for future basic discovery research as well as for at-point-of-care diagnostic or prognostic tests.

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Rapid detection of Newcastle disease virus replication in embryonated chicken eggs using quantitative real time polymerase chain reaction

Cited by 13 publications

References 10 publications

Toll-Like Receptor Responses to Peste des petits ruminants Virus in Goats and Water Buffalo

Toll-Like Receptor Responses to Peste des petits ruminants Virus in Goats and Water Buffalo

Viability Reduction andRac1Gene Downregulation of HeterogeneousEx-VivoGlioma Acute Slice Infected by the Oncolytic Newcastle Disease Virus Strain V4UPM

Strategies for highly sensitive biomarker detection by Rolling Circle Amplification of signals from nucleic acid composed sensors

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