MSACL 2019 EU Abstract

Self-Classified Topic Area(s): Proteins & Proteomics

Single-Shot Targeted Proteomics Flavivirus Assay

Sheena Wee (1), Asfa Alli-Shaik (1), Relus Kek (2), Hannah L. F. Swa (1), Wei-Ping Tien (2), Vanessa W. Lim (3), Yee-Sin Leoc (3, 4), Lee-Ching Ng (2, 5), Hapuarachchige C. Hapuarachchi (2), and Jayantha Gunaratne (1, 6)
(1) Institute of Molecular and Cell Biology, Agency for Science, Technology and Research, Singapore (2) Environmental Health Institute, National Environment Agency, Singapore (3) National Centre for Infectious Diseases, Singapore (4) Communicable Disease Centre, Tan Tock Seng Hospital, Singapore (5) School of Biological Sciences, Nanyang Technological University, Singapore (6) Yong Loo Lin School of Medicine, National University of Singapore, Singapore

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Jayantha Gunaratne (Presenter)
Institute of Molecular and Cell Biology, A*STAR

Presenter Bio: Jayantha Gunaratne obtained his Ph.D. in Biosciences from Tokyo Institute of Technology, Japan in 2003 and was a postdoctoral fellow at University of California San Diego from 2004 to 2007. In 2007, he moved to Singapore as a founder member of the Mass Spectrometry and Systems Biology Laboratory in Institute of Molecular and Cell Biology (IMCB), A*STAR and pioneered the establishment of high-resolution advanced quantitative proteomics technology in Singapore. He is currently a Principal Investigator heading translational biomedical proteomics research in IMCB and also an adjunct Associate Professor in Yong Loo Lin School of Medicine, National University of Singapore.

Relevant Financial Disclosures (within past 24 months)

Salary Institute of Molecular and Cell Biology (My employer)

Conflict mitigation NOT REQUIRED.

Abstract

INTRODUCTION: With pandemic emergence and increasing magnitude of flavivirus outbreaks in recent years, there is an urgent need in the development of a robust diagnostic tool for flavivirus diagnosis and typing to better facilitate disease management, surveillance and control.
OBJECTIVE: The objective of this work was to develop a highly sensitive shotgun diagnostic assay that has typing capability to cover a wide panel of flaviviruses and circumvents the shortcomings of current flavivirus diagnostics assays.
METHODS: Peptides derived from virus and serotype-specific non-structural protein 1 (NS1) were selected as virus/type identifiers to be monitored by targeted proteomics-based parallel reaction monitoring (PRM) approach. This assay was validated using clinical and infected secretome samples.
RESULTS: The assay was shown to confidently distinguish Dengue serotypes (1-4), Zika, Yellow Fever and Kunjin viruses. The ability to diagnose multiple flaviviruses and types over a longer diagnostic window (up to 8 days) with high sensitivity (70% vs 50% by PCR), specificity (100%) and multiplexing capability (7 viruses/types in a single run) in comparison to conventional assays were shown. This assay performs equally well for primary, secondary and co-infections.
CONCLUSIONS: Our flavivirus PRM assay, which can be easily extendable to detect a wider panel of flaviviruses, is the first single-shot flavivirus assay and has addressed the shortcomings of current diagnostics, positioning it to hold high promise as a future flavivirus diagnostic tool.
Reference: Wee et al (2019. Multiplex targeted mass spectrometry assay for one-shot flavivirus diagnosis. Proc Natl Acad Sci U S A. 116:6754-6759