Date of Award

Spring 2000

Document Type

Restricted Thesis

Terms of Use

© 2000 Joseph D. Tucker. All rights reserved. Access to this work is restricted to users within the Swarthmore College network and may only be used for non-commercial, educational, and research purposes. Sharing with users outside of the Swarthmore College network is expressly prohibited. For all other uses, including reproduction and distribution, please contact the copyright holder.

Degree Name

Bachelor of Arts

Department

Biology Department

First Advisor

Amy Cheng Vollmer

Abstract

Hepatitis A Virus (HAV) is an infectious disease spread through the oral-fecal route. Approximately 180,000 Americans acquire hepatitis A infection each year, costing $1,817 - $2,459 per symptomatic case. HAV environmental surveillance in remote locations requires inexpensive, flexible, and sensitive molecular detection methods. Since the virus is endemic among many rural Native American subpopulations, adapting a new method for sustainable rural HAV detection could greatly benefit these high risk areas. This project established a portable HAV reverse transcription PCR (RT-PCR) assay for environmental surveillance from water, sewage, or clam tissue. Mercenaria mercenaria (hard shell clams) concentrate the virus through filter-feeding, providing a natural environmental sentinel for detection. The technique could detect as few as 30-100PFU HAV per liter of water, 1PFU per 10mL of urea-arginine phosphate buffer, and 0.3 PFU HAV per 2mL of guanidium isothiocyanate solution. Clam dissection utilized compartmentalized bivalve physiology to isolate tissues more likely to contain HAV. Environmental detection on the Hoopa Valley Indian Reservation (Northern California) pinpointed a 1L water sample with at least 30PFU of HAV in a localized area of the reservation. Sequencing of the amplicon was consistent with a 1B genotype of HAV, supporting epidemiological evidence. Sustained hepatitis A virus detection alongside community awareness initiatives make another hepatitis A outbreak on the Hoopa Valley Reservation less likely. Combining molecular and community-centered empowerment strategies to respond against infectious disease provides a powerful tool in disease control.

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