Development of a GeXP based multiplex RT-PCR assay for simultaneous detection of eight arboviruses related to encephalitis.
- Author:
Bin HE
1
;
Huan-Yu WANG
;
Chen ZHANG
;
Miao WANG
;
Meng QIN
;
Ke-Xia WANG
;
Xue-Jun MA
Author Information
1. School of Medicine, AnHui University of Science and Technology, Huainan 232001 China.
- Publication Type:Journal Article
- MeSH:
Arboviruses;
genetics;
isolation & purification;
Encephalitis Virus, Japanese;
genetics;
Encephalitis Viruses;
genetics;
isolation & purification;
Reverse Transcriptase Polymerase Chain Reaction;
methods;
Sensitivity and Specificity
- From:
Chinese Journal of Virology
2012;28(1):57-62
- CountryChina
- Language:Chinese
-
Abstract:
Multiplex reverse transcription-polymerase chain reaction (mRT-PCR) is currently available in virus detection and defined as the simultaneous amplification of two or more DNA/RNA targets in a single reaction vessel. In this study, we attempted to modify the conventional mRT-PCR technique on a basis of GenomeLab Genetic Analysis System (GeXP). Initially, we optimized the analytical validation of the GeXP analyzer and its design of workflow and simultaneously detected eight arboviruses that related to epidemic encephalitis by verifying the specificity of mRT-PCR with Japanese encephalitis virus(JEV) cell cultures and positive strains identified previously and determining the sensitivity with in vitro-transcribed RNA of serial dilutions. The GeXP system after optimization could amplify the specific fragments related to the viruses and exposed specifically a total of 13 target genes out of eight types of arboviruses at the level of 10(2) copies/microL, and the findings suggest that the novel protocol we developed can be high-throughput and highly specific and sensitive as well as quickness in screening of the encephalitis viruses, and is promising in detection of encephalitis-associated viruses for molecular epidemiological studies.
