Reverse transcription-PCR-enzyme-linked immunosorbent assay for rapid detection and differentiation of alphavirus infections.

Reverse transcription-PCR-enzyme-linked immunosorbent assay for rapid detection and differentiation of alphavirus infections. Research Abstract Details 

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Reverse transcription-PCR-enzyme-linked immunosorbent assay for rapid detection and differentiation of alphavirus infections. Abstract Text:

Eryu Wang,Slobodan Paessler,Patricia V Aguilar,Anne-Sophie Carrara,Haolin Ni,Ivorlyne P Greene,Scott C Weaver,

Due to the lack of a rapid, simple, and inexpensive assay for detecting alphavirus infections, we combined a reverse transcription-PCR with an enzyme-linked immunosorbent assay (RT-PCR-ELISA) to identify human pathogenic alphaviruses that are endemic in the New World. By combining the sensitivity of PCR, the detection simplicity of ELISA, and the specificities of DNA probes, this method rapidly detected and differentiated closely related species and subtypes of several medically important alphaviruses. After an amplification using RT-PCR with primers targeting conserved sequences in the nonstructural protein 1 gene, sequence-specific, biotin-labeled probes targeted against Venezuelan, eastern, and western equine encephalitis or Mayaro virus genes were used for the detection of amplicons using ELISA. The assay is simple, fast, and easy to perform in an ordinary diagnostic laboratory or clinical setting. Nucleic acid derived from cell cultures infected with several alphaviruses, clinical specimens, and mosquito pools as well as frozen and paraffin-embedded animal tissues were detected and identified within 6 to 7 h in a sensitive and specific manner.

Reverse transcription-PCR-enzyme-linked immunosorbent assay for rapid detection and differentiation of alphavirus infections. Publishing Authors By Initials

E Wang,S Paessler,PV Aguilar,AS Carrara,H Ni,IP Greene,SC Weaver,

For similar investigative techniques: epidemiologic methods: statistics as topic: sensitivity and specificity research abstracts see: investigative techniques: epidemiologic methods: statistics as topic: sensitivity and specificity research

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Reverse transcription-PCR-enzyme-linked immunosorbent assay for rapid detection and differentiation of alphavirus infections. Journal Published:

PUBLICATION TYPE: Research Support, U.S. Gov't,

Journal: Journal of clinical microbiology

VOLUME: 44

Page Numbers: 4000-8

Journal Abbreviation: J. Clin. Microbiol.

ISSN: 0095-1137

DAY: 6

MONTH: 09

YEAR: 2006

Reverse transcription-PCR-enzyme-linked immunosorbent assay for rapid detection and differentiation of alphavirus infections. Information

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LANGUAGE: eng

NlmUniqueID: 7505564

Reverse transcription-PCR-enzyme-linked immunosorbent assay for rapid detection and differentiation of alphavirus infections. Keywords Mesh Terms:

KEYWORDS: Sensitivity and Specificity

MESH TERMS: methods

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Grant and Affiliation Information for Reverse transcription-PCR-enzyme-linked immunosorbent assay for rapid detection and differentiation of alphavirus infections.

AFFILIATION: Center for Biodefense and Emerging Infectious Diseases, Department of Pathology, University of Texas Medical Branch, Galveston, Texas 77555-0609, USA.

Country: United States

AGENCY: United States NIAID

GRANT: U54 AI057156

ACRONYM: AI

MEDLINETA: J Clin Microbiol

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