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Isolation of Rickettsia amblyommatis in HUVEC line

S Santibáñez1, A Portillo1, A M Palomar1

  • 1Center of Rickettsiosis and Arthropod-Borne Diseases, Infectious Diseases Department, Hospital San Pedro-Center of Biomedical Research from La Rioja (CIBIR), Logroño, La Rioja, Spain.

New Microbes and New Infections
|January 12, 2018
PubMed

Insights

Rickettsia amblyommatis, a common tick bacterium, was successfully isolated and cultured in human umbilical vein endothelial cells (HUVEC). This study demonstrates HUVEC

Area of Science:

  • Microbiology
  • Infectious Diseases
  • Vector-borne Diseases

Background:

  • Rickettsia amblyommatis is a prevalent intracellular bacterium in the spotted fever group Rickettsia, commonly found in Amblyomma ticks across the Western Hemisphere.
  • Previous studies have cultivated R. amblyommatis in various cell types, including chicken fibroblasts, embryonated eggs, and Vero cells.

Purpose of the Study:

  • To isolate Rickettsia amblyommatis from Amblyomma cajennense tick using human umbilical vein endothelial cells (HUVEC).
  • To establish an in vitro culture of R. amblyommatis in HUVEC for further study and characterization.

Main Methods:

  • A nymph of Amblyomma cajennense was processed, with one half used for ompA PCR and the other for in vitro culture.
  • Tick homogenate was inoculated into a HUVEC line and incubated at 33°C with specific growth medium.
  • Cultures were monitored weekly using Gimenez staining, and PCR and sequencing were performed on observed Rickettsia-like organisms.

Main Results:

  • PCR analysis of the tick half confirmed the presence of R. amblyommatis.
  • After 48 days of incubation, Rickettsia-like organisms were observed intracellularly within HUVEC.
  • Sequencing of the ompA gene from the cultured organisms showed 100% identity with R. amblyommatis.

Conclusions:

  • Human umbilical vein endothelial cells (HUVEC) are an effective tool for the isolation and in vitro cultivation of Rickettsia amblyommatis.
  • The successful isolation and culture of R. amblyommatis in HUVEC provide a valuable resource for studying this important tick-borne bacterium.
  • The established R. amblyommatis isolate has been deposited in a recognized culture collection for future research.
Keywords:
Amblyomma cajennenseCandidatus Rickettsia amblyommiiHUVEC lineRickettsia amblyommatis

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