Graduation Year


Document Type




Degree Name

MS in Public Health (M.S.P.H.)

Degree Granting Department

Global Health

Major Professor

Thomas Unnasch, Ph.D.

Co-Major Professor

Miguel Reina-Ortiz, Ph.D.

Committee Member

Alexis LaCrue, Ph.D.

Committee Member

Andreas Seyfang, Ph.D.

Committee Member

Lea Heberlein-Larson, MPH


Plaque reduction neutralization test, Chimera viruses, Arboviruses


The plaque reduction neutralization test (PRNT) is a confirmatory diagnostic assay that is used to confirm a variety of diseases. The performance of PRNT requires the use of infectious wild type viruses, which increases the risk of laboratory acquired infections. For instance, eastern equine encephalitis (EEEV) is a highly virulent pathogen used in PRNT that can result in potentially fatal neurological diseases among humans and equines. Therefore, arboviral PRNT must be performed in Biosafety Level 3 (BSL-3) containment facilities and may require select agent approved scientists, like in the case of EEEV. These stringent requirements restrict the ability of public health laboratories to conduct PRNTs. Chimera viruses, recombinant constructs that have been bio-engineered to express the immunogenic structural proteins from the wild type virus in an attenuated form, can serve as a substitution for infectious viruses when performing PRNT. Since chimera viruses do not require the use of a BSL-3 facility and are not classified as select agents, their use offers advantages over wild type viruses. This study aimed at validating the use of EEE and West Nile chimera viruses as an alternative to the corresponding wild type viruses for diagnostic purposes at the Florida Department of Health (FDOH) Bureau of Public Health Laboratories (BPHL). These evaluations were conducted using human and avian sera. The results illustrate that chimera virus-based PRNT portrays specificity comparable to that of the wild type virus, while a slight reduction in sensitivity was observed when human sera was used. Considering their benefits in increasing safety and reducing regulatory requirements, these chimera viruses are an important alternative to the virulent wild type viruses and could be highly beneficial for diagnostic laboratories.

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