Shell-vial culture, coupled with real-time PCR, applied to Rickettsia conorii and Rickettsia massiliae-Bar29 detection, improving the diagnosis of the Mediterranean spotted fever

Ferran Segura, Immaculada Pons, Isabel Sanfeliu, María Mercedes Nogueras

Research output: Contribution to journalArticleResearchpeer-review

3 Citations (Scopus)

Abstract

© 2016 Elsevier GmbH. Rickettsia conorii and Rickettsia massiliae-Bar29 are related to Mediterranean spotted fever (MSF). They are intracellular microorganisms. The Shell-vial culture assay (SV) improved Rickettsia culture but it still has some limitations: blood usually contains low amount of microorganisms and the samples that contain the highest amount of them are non-sterile. The objectives of this study were to optimize SV culture conditions and monitoring methods and to establish antibiotic concentrations useful for non-sterile samples. 12 SVs were inoculated with each microorganism, incubated at different temperatures and monitored by classical methods and real-time PCR. R. conorii was detected by all methods at all temperatures since 7th day of incubation. R. massiliae-Bar29 was firstly observed at 28 °C. Real-time PCR allowed to detected it 2-7 days earlier (depend on temperature) than classical methods. Antibiotics concentration needed for the isolation of these Rickettsia species from non-sterile samples was determined inoculating SV with R. conorii, R. massiliae-Bar29, biopsy or tick, incubating them with different dilutions of antibiotics and monitoring them weekly. To sum up, if a MSF diagnosis is suspected, SV should be incubated at both 28 °C and 32 °C for 1-3 weeks and monitored by a sensitive real-time PCR. If the sample is non-sterile the panel of antibiotics tested can be added.
Original languageEnglish
Pages (from-to)457-461
JournalTicks and Tick-borne Diseases
Volume7
Issue number3
DOIs
Publication statusPublished - 1 Apr 2016

Keywords

  • Mediterranean spotted fever
  • Real-time PCR
  • Rickettsia conorii
  • Rickettsia massiliae-Bar29
  • Shell-vial assay

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