Abstract：
:Porcine reproductive and respiratory syndrome virus (PRRSV) became a significant pathogen of swine upon its emergence in the late 1980s and since then has exemplified a rapidly evolving, constantly re-emerging pathogen. In addition to the challenges faced in development of vaccines and diagnostics, research on the basic molecular pathogenesis of PRRSV is also restrained by the ability to accurately and comparatively quantitate levels of replication in different tissues and between strains. This is further complicated by the presence of non-genomic RNA within infected tissues which are generally detected with equivalent efficiency by RT-qPCR based techniques, thereby introducing inherent error in these measurements that may differ significantly by tissue and strain. To address this, an RT-qPCR based technique was developed which targets the viral RNA-dependent RNA polymerase gene (nsp9) which is unique to genomic RNA, being absent from all subgenomic and heteroclite RNAs. This assay targets a region of considerable sequence conservation, and based on sequence only, should be quantitative for approximately 40% of all Type 2 PRRSV strains in GenBank for which nsp9 sequence is available. The assay was demonstrated to be linear over nine orders of magnitude (10(10)-10(2) copies) and can be readily adapted for multiplex detection of additional divergent PRRSV strains. This assay will add significantly to the ability to assess and compare PRRSV replication in a variety of tissues and between divergent strains, including highly pathogenic strains of considerable concern to the global pork industry.
journal_name
J Virol Methodsjournal_title
Journal of virological methodsauthors
Spear A,Faaberg KSdoi
10.1016/j.jviromet.2015.02.007subject
Has Abstractpub_date
2015-06-15 00:00:00pages
1-6eissn
0166-0934issn
1879-0984pii
S0166-0934(15)00029-4journal_volume
218pub_type
杂志文章abstract：:Polymerase chain reaction (PCR) assays were developed for the detection of RNA from the St. Louis encephalitis (SLE) virus. Using computer-assisted analysis of the MSI-7 strain SLE virus genome, two primer pairs were selected from the capsid-coding and the membrane associated protein-coding genes, and one from the env...
journal_title：Journal of virological methods
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journal_title：Journal of virological methods
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journal_title：Journal of virological methods
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journal_title：Journal of virological methods
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journal_title：Journal of virological methods
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journal_title：Journal of virological methods
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journal_title：Journal of virological methods
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pub_type： 杂志文章
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journal_title：Journal of virological methods
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更新日期：2009-07-01 00:00:00
abstract：:A rapid and effective ELISA for measuring Epstein-Barr virus (EBV)-neutralizing antibodies in human sera was devised to replace the existing cumbersome method involving the inhibition of fetal cord blood B-cell transformation by the virus. The new method will be invaluable for assessing antibody responses in human sub...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/s0166-0934(98)00054-8
更新日期：1998-07-01 00:00:00
