Establishment of a high-throughput respiratory virus detection technology without RNA purification and reverse transcription.
- Author:
Dan-li YANG
1
;
Xiao-yi TIAN
;
Wei-xian SHI
;
Zhi ZHENG
Author Information
- Publication Type:Journal Article
- MeSH: High-Throughput Screening Assays; methods; Influenza A virus; genetics; isolation & purification; Influenza B virus; genetics; isolation & purification; Limit of Detection; Metapneumovirus; genetics; isolation & purification; Nucleic Acid Amplification Techniques; methods; RNA, Viral; analysis; Respiratory Syncytial Viruses; genetics; isolation & purification; Respiratory System; virology; Respirovirus; genetics; isolation & purification; Reverse Transcription; Sensitivity and Specificity
- From: Acta Academiae Medicinae Sinicae 2013;35(1):24-28
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo establish a convenient and high-throughput respiratory virus detection method to facilitate epidemiological viral monitoring.
METHODSWe used high-throughput microsphere-based flexible multi-analyte profiling technology (xMAP) coupled with signal amplification molecules to simultaneously detect RNAs of 8 viruses including influenza viruses A and B, parainfluenza viruses type 1, 2 and 3, respiratory syncytial viruses A and B, and metapneumovirus in a 96-well plate format. The sensitivity and specificity of the method for the synthetic viral RNAs were evaluated.
RESULTSThere was no cross-reactivity among the 8 respiratory viral target RNAs. The detection limits for the 8 viral in intro-transcribed RNAs ranged from 1204 to 4695 RNA copies.
CONCLUSIONWe establish a sensitive, specific, convenient, and high-throughput multiplex detection method suitable for detecting multiple respiratory viral RNAs for epidemiological viral monitoring.
