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Compositions and methods for detecting certain flaviviruses, including members of the japanese encephalitis virus serogroup

An encephalitis virus, serum technology, applied in biochemical equipment and methods, microbial assay/test, resistance to vector-borne diseases, etc., can solve problems such as difficulty in distinguishing between active infection and previous contact

Active Publication Date: 2006-06-28
F HOFFMANN LA ROCHE & CO AG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, WNV-specific IgM persists for more than 1 year, making it difficult to distinguish between active infection and previous exposure

Method used

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  • Compositions and methods for detecting certain flaviviruses, including members of the japanese encephalitis virus serogroup
  • Compositions and methods for detecting certain flaviviruses, including members of the japanese encephalitis virus serogroup
  • Compositions and methods for detecting certain flaviviruses, including members of the japanese encephalitis virus serogroup

Examples

Experimental program
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Embodiment 1

[0378] Embodiment 1: Amplification and detection of West Nile virus RNA

[0379] Lysates of virus-infected cell culture supernatants were obtained from Dr. R. Lanciotti, Centers for Disease Control and Prevention. Nucleic acids were purified from the above lysates using reagents in the QIAamp Viral RNA Mini Kit (Qiagen Inc., Valencia, CA) according to the manufacturer's instructions. Prepare 10-fold serial dilutions (10 -2 -10 -7 ). Utilize TaqMan ® reagent and method to carry out 5 ' nuclease reaction test, and by containing 1 μ M primer (including SEQ ID NO:8 and SEQ ID NO:15), 55mM Tricine (pH7.7, Sigma, cat T-5816), 450 μM dNTPs (including dATP, dCTP, dGTP and dUTP, Pharmacia), 2.7 mM manganese acetate (Fluka, cat 63537), 135 mM potassium acetate (Fluka, cat 60035), 7% (v / v) DMSO (Sigma, cat D8418) , 6% (V / V) glycerol (USB, cat 16347), 5 units of uracil-N-glycosylase (Roche Diagnostics), 40 units of Z05 DNA polymerase (Roche Diagnostics) and 0.15 μM probe (SEQ ID NO: ...

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Abstract

The present invention provides rapid and accurate methods, primers, probes and kits for detecting certain flaviviruses in samples. Detectable flaviviruses include Japanese encephalitis virus serogroup members, dengue virus, St. Louis encephalitis virus, Montana myotis leukoencephalitis virus, Modoc virus, and yellow fever virus. The primers and probes of the present invention can hybridize to the region within the 3' untranslated region of the genome of the virus to be detected.

Description

Background of the invention [0001] The Flaviviridae family and the Flavivirus genus include many viruses that are potentially lethal pathogens in humans. These viruses include dengue virus, yellow fever virus, Modoc virus, and viruses in the Japanese encephalitis virus serogroup. The Japanese encephalitis virus serogroup includes several closely related viruses such as Japanese encephalitis virus (JEV), West Nile virus (WNV), St. Louis encephalitis virus, Murray Valley encephalitis virus, and Kunjin virus. The reason Kunjin viruses are often considered WNV variants is the degree of sequence conservation between the two viruses. According to the results of sequence analysis, the characterized WNV strains can be divided into two groups, namely lineage I and lineage II. The first human WNV infection in the United States was reported in 1999. Since then, it has exploded every year. In August 2002, when 4 organ recipients were infected by a single organ donor, it was confirmed ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/70
CPCY02A50/30
Inventor K·K·Y·杨
Owner F HOFFMANN LA ROCHE & CO AG
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