Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

Outer Membrane Protein A Conservation among Orientia tsutsugamushi Isolates Suggests Its Potential as a Protective Antigen and Diagnostic Target

Version 1 : Received: 11 April 2018 / Approved: 11 April 2018 / Online: 11 April 2018 (08:13:04 CEST)

A peer-reviewed article of this Preprint also exists.

Evans, S.M.; Adcox, H.E.; VieBrock, L.; Green, R.S.; Luce-Fedrow, A.; Chattopadhyay, S.; Jiang, J.; Marconi, R.T.; Paris, D.; Richards, A.L.; Carlyon, J.A. Outer Membrane Protein A Conservation among Orientia tsutsugamushi Isolates Suggests Its Potential as a Protective Antigen and Diagnostic Target. Trop. Med. Infect. Dis. 2018, 3, 63. Evans, S.M.; Adcox, H.E.; VieBrock, L.; Green, R.S.; Luce-Fedrow, A.; Chattopadhyay, S.; Jiang, J.; Marconi, R.T.; Paris, D.; Richards, A.L.; Carlyon, J.A. Outer Membrane Protein A Conservation among Orientia tsutsugamushi Isolates Suggests Its Potential as a Protective Antigen and Diagnostic Target. Trop. Med. Infect. Dis. 2018, 3, 63.

Abstract

Scrub typhus threatens one billion people in the Asia-Pacific area and cases have emerged outside this region. It is caused by infection with any of the multitude of strains of the bacterium, Orientia tsutsugamushi. A vaccine that affords heterologous protection and a commercially available molecular diagnostic assay are lacking. Herein, we determined that the nucleotide and translated amino acid sequences of outer membrane protein A (OmpA) are highly conserved among 51 O. tsutsugamushi isolates. Molecular modeling revealed the predicted tertiary structure of O. tsutsugamushi OmpA to be very similar to that of the phylogenetically related pathogen, Anaplasma phagocytophilum, including the location of a helix that contains residues functionally essential for A. phagocytophilum infection. PCR primers were developed that amplified ompA DNA from all O. tsutsugamushi strains, but not from negative control bacteria. Using these primers in quantitative real-time PCR enabled sensitive detection and quantitation of O. tsutsugamushi ompA DNA from organs of mice that had been experimentally infected with the Karp or Gilliam strains. The high degree of OmpA conservation among O. tsutsugamushi strains evidences its potential to serve as a molecular diagnostic target and justifies its consideration as a candidate for developing a broadly protective scrub typhus vaccine.

Keywords

scrub typhus; Orientia tsutsugamushi; Rickettsia; Rickettsiales; outer membrane protein A; Anaplasma

Subject

Biology and Life Sciences, Immunology and Microbiology

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