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Multiplexed Isothermal Amplification Based Diagnostic Platform to Detect Zika, Chikungunya, and Dengue 1
Published on: March 13, 2018
Development and evaluation of serotype- and group-specific fluorogenic reverse transcriptase PCR (TaqMan) assays for
J D Callahan1, S J Wu, A Dion-Schultz
1Viral and Rickettsial Diseases Department, Naval Medical Research Center, Silver Spring, Maryland 20910-7500, USA.
Journal of Clinical Microbiology
|October 30, 2001
Summary
Developed rapid dengue virus detection assays using TaqMan reverse transcriptase PCR (RT-PCR). These sensitive and specific tools enable quick serotyping and screening for epidemiological studies of dengue virus infections.
Area of Science:
- Molecular Biology
- Virology
- Infectious Diseases
Background:
- Dengue virus poses a significant global health threat, necessitating rapid and accurate diagnostic tools.
- Existing diagnostic methods can be time-consuming and may lack serotype specificity.
Purpose of the Study:
- To develop and evaluate fluorogenic probe hydrolysis (TaqMan) reverse transcriptase PCR (RT-PCR) assays for dengue virus detection.
- To achieve both serotype-specific and group-specific identification of dengue virus strains.
Main Methods:
- Design of serotype- and group-specific oligonucleotide primers and fluorogenic probes targeting conserved dengue virus genome regions.
- Development of a rapid, single-tube RT-PCR assay with a 30-minute reverse transcription step and a 45-cycle PCR.
- Validation of assays using cell culture-derived viruses and 67 dengue-viremic human sera from diverse geographical locations.
Main Results:
- The serotype-specific TaqMan RT-PCR assay demonstrated 92.5% sensitivity in detecting dengue virus.
- The group-specific TaqMan RT-PCR assay achieved 98.5% sensitivity.
- All TaqMan RT-PCR assays exhibited 100% specificity when compared to cell culture isolation and testing of negative control samples.
Conclusions:
- The developed TaqMan RT-PCR assays are rapid, sensitive, and specific for dengue virus detection.
- These assays can serve as valuable tools for epidemiological surveillance and diagnosis of dengue virus infections.
- The method facilitates efficient screening and serotyping, aiding in the understanding and control of dengue outbreaks.
