Genetic typing of isolates of Rickettsia typhi

Author:

Kato Cecilia Y.ORCID,Chung Ida H.ORCID,Robinson Lauren K.ORCID,Eremeeva Marina E.,Dasch Gregory A.ORCID

Abstract

Murine typhus, which is caused byRickettsia typhi, has a wide range of clinical manifestations. It has a low mortality rate but may result in meningoencephalitis and interstitial pneumonia in severe cases. Comparisons of complete genome sequences ofR.typhiisolates from North Carolina, USA (Wilmington), Myanmar (B9991PP), and Thailand (TH1527) identified only 26 single nucleotide polymorphism (SNP) and 7 insertion-deletion (INDEL) sites in these highly syntenic genomes. Assays were developed to further define the distribution of these variant sites among 15 additional isolates ofR.typhiwith different histories from Asia, the USA, and Africa. Mismatch amplification mutation assays (MAMA) were validated for 22 SNP sites, while the 7 INDEL sites were analyzed directly on agarose gels. Six SNP types, 9 INDEL types, 11 total types were identified among these 18 isolates. Replicate DNA samples as well as comparisons of isolates with different passage and source histories gave consistent genetic typing profiles. Comparison of the SNP and INDEL markers toR.typhi’snearest neighborRickettsia prowazekiidemonstrated that the majority of the SNPs represent intra-species variation that arose post divergence of these two species while several INDEL sites also exhibited intraspecies variability among theR.prowazekiigenomes that have been completely sequenced. The assays for the presence of these SNP and INDEL sites, particularly the latter, comprise a low technology gel method for consistently distinguishingR.typhi and R.prowazekiias well as for differentiating genetic types ofR.typhi.

Publisher

Public Library of Science (PLoS)

Subject

Infectious Diseases,Public Health, Environmental and Occupational Health

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