Development of Q-PCR Technique for Determining Bartonella Infection in Wild Deer Mice (Peromyscus maniculatus)

Burns, Amelia and McLean, Nellie and Paddock, Sherry A. and Simpkiss, Alena G. and Sommerville , Dr. Les (2012) Development of Q-PCR Technique for Determining Bartonella Infection in Wild Deer Mice (Peromyscus maniculatus). [Abstract]

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Abstract

Sin Nombre virus (SNV) is a North American hantavirus that causes Hantavirus Pulmonary Syndrome (HPS) in humans, an acute and rapidly progressing disease with high (~40%) mortality rates. Deer mice (Peromyscus maniculatus) are the primary mode of human HPS infection; humans become infected with SNV via inhalation of virus particles shed through deer mouse excrement. It is unknown whether chronic infection with SNV impacts the ability of deer mice to respond to other immune challenges. This has significant implications for the deer mouse – SNV system, as deer mice with reduced immunocompetence may shed greater amounts of SNV, increasing the likelihood of human SNV infection. Thus, our objective was to develop a protocol in which we could quantify Bartonella, a bacterial pathogen that is endemic in wild rodents, through Q-PCR. Similar to SNV, Bartonella has low pathogenicity, but requires that deer mice mount a chronic, low-level immune response post infection. For the development of a Q-PCR assay, we are using the forward and reverse primers BhC5781p (5’-GGGGACCAGCTCATGGTGG-3’) and BhCS1137n, (5’-AATGCAAAAAGAACAGTAAACA-3’ respectively. Q-PCR trials were completed using serum and whole blood, however, we were unable to isolate and amplify Bartonella DNA using serum and whole blood alone. DNA isolation appears to be a necessary step to detect the presence of Bartonella DNA in the deer mouse whole blood samples. The further development of this protocol will allow us to determine the extent to which SNV and Bartonella infections are correlated. This research will ultimately improve our ability to predict human patterns of infection with zoonotic pathogens.

Item Type: Abstract
Created by Student or Faculty: Student
Additional Information: 7th Annual Natural & Behavioral Sciences Undergraduate Research Symposium Program
Uncontrolled Keywords: Sin Nombre virus (SNV), Hantavirus, Hantavirus Pulmonary Syndrome (HPS), Mortality, Deer mice (Peromyscus maniculatus), Bartonella, Q-PCR technique, pathogenicity, BhC5781p (5’-GGGGACCAGCTCATGGTGG-3’), BhCS1137n, (5’-AATGCAAAAAGAACAGTAAACA-3’, DNA isolation, zoonotic pathogens, Infection, Infectious diseases; Transmission patterns;
Subjects: School of Natural and Behavioral Sciences > Chemistry
School of Natural and Behavioral Sciences > Environmental Studies
NBS Symposium
Interdisciplinary > Pre-Health
School of Natural and Behavioral Sciences > Public Health
Depositing User: Alejandro Marquez
Date Deposited: 26 Jun 2013 14:13
Last Modified: 26 Jun 2013 14:13
URI: http://eprints.fortlewis.edu/id/eprint/279


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