Date of Award

January 2022

Document Type

Open Access Thesis

Degree Name

Master of Public Health (MPH)

Department

School of Public Health

First Advisor

Elsio Wunder Jr.

Abstract

Leptospirosis is an infectious zoonotic disease caused by bacteria of the genus Leptospira. The genus is comprised of 40 pathogenic species divided among P1 (19) and P2 (21). Leptospirosis incurs a disproportionately large burden of disease in urban slum regions of developing countries that lack adequate sewage disposal, water treatment, and sanitation infrastructure. While P1 species have been identified in cases of human infection, it is unclear if all Leptospira species are capable of infecting humans, especially P2 species. Among P1 species, L. interrogans has been described as the most frequent specie to cause disease in both animals and humans, and varying pathogenicity has been identified in other pathogenic species. Real-time qPCR assays have been shown to identify Leptospira with high sensitivity and specificity; however, there is a lack of assays capable of targeting relevant species of Leptospira of specific clinical and environmental importance among both humans and animals. Thus, the purpose of this study was three-fold: (1) design a multiplex assay for the simultaneous detection of and discrimination between P1 and P2 species; (2) optimize a previous assay for the detection of all currently-named P1 species; and (3) create an assay for the specific detection of L. interrogans. Primers and probes were designed in silico utilizing the BioEdit software to evaluate sequences of all 68 species with available genomic sequences. In vitro sensitivity and specificity analyses were conducted on 28 reference species. Given the specificity and sensitivity of the developed assays, real-time qPCR assays may be useful in future epidemiological and clinical studies for the identification and quantification of various Leptospira species.

Comments

This is an Open Access Thesis.

Download
Open Access

This Article is Open Access

Share

COinS