Isolation of in HUVEC line.
Journal:
New microbes and new infections
Published Date:
Dec 9, 2017
Abstract
formerly named and ' Rickettsia amblyommii' is an intracellular bacterium belonging to the spotted fever group It is highly prevalent in and in other spp. throughout the Western Hemisphere. has been cultivated in chicken fibroblast, primary embryonated chicken eggs, Vero cells and arthropod-derived cells. Because of the affinity of rickettsiae to invade vascular endothelial cells, we tried to isolate from a nymph of s.l. collected in Saltillo (Coahulia, Mexico) using human umbilical vein endothelial cells (HUVEC). One tick half was analysed by PCR and was found to be positive for The other half was selected for culture of spp. It was triturated in 1 mL of endothelial cell growth medium with 1% antibiotic-antimycotic solution, and the homogenate was inoculated into a HUVEC line. Culture was maintained at 33°C in endothelial cell growth medium plus 2 mM l-glutamine and 2% fetal calf serum, with 5% CO The medium was changed weekly. Culture was checked by Gimenez stain for -like intracellular organisms. After 48 days of incubation, -like organisms were observed in HUVEC. PCR assays and sequencing of gene in the culture suspension showed 100% identity with This isolate was successfully established in HUVEC, and it has been deposited in the collection of the Center of Rickettsioses and Arthropod-Borne Diseases, Infectious Diseases Department, Hospital San Pedro-Center of Biomedical Research from La Rioja, Logroño, Spain. The HUVEC line is a useful tool for the isolation of
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