The genus Rickettsia (Rickettsiales: Rickettsiaceae) includes Gram-negative, small, obligate intracellular, nonmotile, pleomorphic coccobacilli bacteria transmitted by arthropods. Some of them cause human and probably also animal disease (life threatening in some patients). In these guidelines, we give clinical practice advices (microscopy, serology, molecular tools, and culture) for the microbiological study of these microorganisms in clinical samples. Since in our environment rickettsioses are mainly transmitted by ticks, practical information for the identification of these arthropods and for the study of Rickettsia infections in ticks has also been added.
Get full access to this article
View all access options for this article.
References
1.
AlexandreN, SantosAS, BacellarF, BoinasFJ, et al.Detection of Rickettsia conorii strains in Portuguese dogs (Canis familiaris). Ticks Tick Borne Dis, 2011; 2:119–122.
2.
AngelakisE, RichetH, RolainJM, La ScolaB, et al.Comparison of real-time quantitative PCR and culture for the diagnosis of emerging Rickettsioses. PLoS Negl Trop Dis, 2012; 6:e1540.
3.
BalrajP, VestrisG, RaoultD, RenestoP. Comparison of Rickettsia conorii growth within different cell lines by real-time quantitative PCR. Clin Microbiol Infect, 2009; 15(Suppl 2):294–295.
4.
BechahY, SocolovschiC, RaoultD. Identification of rickettsial infections by using cutaneous swab specimens and PCR. Emerg Infect Dis, 2011; 17:83–86.
5.
Bell-SakyiL. Ehrlichia ruminantium grows in cell lines from four ixodid tick genera. J Comp Pathol, 2004; 130:285–293.
6.
Bell-SakyiL, ZweygarthE, BlouinEF, GouldEA, et al.Tick cell lines: Tools for tick and tick-borne disease research. Trends Parasitol, 2007; 23:450–457.
7.
BrouquiP, BacellarF, BarantonG, BirtlesJ, et al.Guidelines for the diagnosis of tick-borne diseases in Europe. Clin Microbiol Infect, 2004; 10:1108–1132.
8.
ChoiYJ, JangWJ, KimJY, RyuJS, et al.Spotted fever group and typhus group rickettsioses in humans, South Korea. Emerg Infec Dis, 2005; 11:237–244.
9.
De SousaR, IsmailN, Dória-NóbregaS, CostaP, et al.The presence of eschars, but not greater severity, in Portuguese patients infected with Israeli spotted fever. Ann N Y Acad Sci, 2005; 1063:197–202.
10.
DenisonAM, AminBD, NicholsonWL, PaddockCD. Detection of Rickettsia rickettsii, Rickettsia parkeri, and Rickettsia akari in skin biopsy specimens using a multiplex real-time polymerase chain reaction assay. Clin Infect Dis, 2014; 59:635–642.
11.
DiemeC, BechahY, SocolovschiC, AudolyG, et al.Transmission potential of Rickettsia felis infection by Anopheles gambiae mosquitoes. Proc Natl Acad Sci U S A, 2015; 112:8088–8093.
12.
DumlerJS, GageWR, PettisGL, AzadAF, et al.Rapid immunoperoxidase demonstration of Rickettsia rickettsii in fixed cutaneous specimens from patients with Rocky Mountain spotted fever. Am J Clin Pathol, 1990; 93:410–414.
13.
Faccini-MartínezÁA, García-ÁlvarezL, HidalgoM, OteoJA. Syndromic classification of rickettsioses: An approach for clinical practice. Int J Infect Dis, 2014; 28:126–139.
14.
FournierPE, JenseniusM, LaferlH, VeneS, et al.Kinetics of antibody responses in Rickettsia africae and Rickettsia conorii infections. Clin Diagn Lab Immunol, 2002; 9:324–328.
15.
FournierPE, RaoultD. Suicide PCR on skin biopsy specimens for diagnosis of rickettsioses. J Clin Microbiol, 2004; 42:3428–3434.
16.
GourietF, FenollarF, PatriceJY, DrancourtM, et al.Use of shell-vial cell culture assay for isolation of bacteria from clinical specimens: 13 years of experience. J Clin Microbiol, 2005; 43:4993–5002.
17.
HortaMC, LabrunaMB, DurigonEL, SchumakerTT. Isolation of Rickettsia felis in the mosquito cell line C6/36. Appl Environ Microbiol, 2006; 72:1705–1707.
18.
JadoI, EscuderoR, GilH, Jiménez-AlonsoMI, et al.Molecular method for identification of Rickettsia species in clinical and environmental samples. J Clin Microbiol, 2006; 44:4572–4576.
19.
KiddL, MaggiR, DinizPP, HegartyB, et al.Evaluation of conventional and real-time PCR assays for detection and differentiation of spotted fever group Rickettsia in dog blood. Vet Microbiol, 2008; 129:294–303.
20.
La ScolaB, RaoultD. Diagnosis of Mediterranean spotted fever by cultivation of Rickettsia conorii from blood and skin samples using the centrifugation-shell vial technique and by detection of R. conorii in circulating endothelial cells: A 6-year follow-up. J Clin Microbiol, 1996; 34:2722–2727.
21.
La ScolaB, RaoultD. Laboratory diagnosis of rickettsioses: Current approaches to diagnosis of old and new rickettsial diseases. J Clin Microbiol, 1997; 35:2715–2727.
22.
La ScolaB, RydkinaL, NdihokubwayoJB, VeneS, et al.Serological differentiation of murine typhus and epidemic typhus using cross-adsorption and Western blotting. Clin Diagn Lab Immunol, 2000; 7:612–616.
23.
LabrunaMB, PachecoRC, NavaS, BrandãoPE, et al.Infection by Rickettsia bellii and Candidatus “Rickettsia amblyommii” in Amblyomma neumanni ticks from Argentina. Microb Ecol, 2007; 54:126–133.
24.
LagierJC, EdouardS, PagnierI, MediannikovO, et al.Current and past strategies for bacterial culture in clinical microbiology. Clin Microbiol Rev, 2015; 28:208–236.
25.
LepidiH, FournierPE, RaoultD. Histologic features and immunodetection of African tick bite fever eschar. Emerging Infect Dis, 2006; 12:1332–1337.
26.
MárquezFJ, MuniainMA, SoriguerRC, IzquierdoG, et al.Genotypic identification of an undescribed spotted fever group Rickettsia in Ixodes ricinus from southwestern Spain. Am J Trop Med Hyg, 1998; 58:570–577.
27.
MarreroM, RaoultD. Centrifugation-shell vial technique for rapid detection of Mediterranean spotted fever Rickettsia in blood culture. Am J Trop Med Hyg, 1989; 40:197–199.
28.
MerhejV, AngelakisE, SocolovschiC, RaoultD. Genotyping, evolution and epidemiological findings of Rickettsia species. Infect Genet Evol, 2014; 25:122–137.
29.
MilhanoN, de CarvalhoIL, AlvesAS, ArroubeS, et al.Coinfections of Rickettsia slovaca and Rickettsia helvetica with Borrelia lusitaniae in ticks collected in a Safari Park, Portugal. Ticks Tick Borne Dis, 2010; 1:172–177.
30.
MunderlohUG, JauronSD, FingerleV, LeitritzL, et al.Invasion and intracellular development of the human granulocytic ehrlichiosis agent in tick cell culture. J Clin Microbiol, 1999; 37:2518–2524.
31.
NgwamidibaM, BlancG, RaoultD, FournierPE. Sca1, a previously undescribed paralog from autotransporter protein-encoding genes in Rickettsia species. BMC Microbiol, 2006; 6:12.
32.
PaddockCD, FinleyRW, WrightCS, RobinsonHN, et al.Rickettsia parkeri rickettsiosis and its clinical distinction from Rocky Mountain spotted fever. Clin Infect Dis, 2008; 47:1188–1196.
33.
PaddockCD, GreerPW, FerebeeTL, SingletonJJr, et al.Hidden mortality attributable to Rocky Mountain spotted fever: Immunohistochemical detection of fatal, serologically unconfirmed disease. J Infect Dis, 1999; 179:1469–1476.
34.
ParolaP, PaddockCD, RaoultD. Tick-borne rickettsioses around the world: Emerging diseases challenging old concepts. Clin Microbiol Rev, 2005; 18:719–756.
35.
ParolaP, PaddockCD, SocolovschiC, LabrunaMB, et al.Update on tick-borne rickettsioses around the world: A geographic approach. Clin Microbiol Rev, 2013; 26:657–702.
36.
PortilloA, SantibáñezS, García-ÁlvarezL, PalomarAM, et al.Rickettsioses in Europe. Microbes Infect. 2015; 17:834–838.
37.
RaoultD, FournierPE, EremeevaM, GravesS, et al.Naming of Rickettsiae and rickettsial diseases. Ann N Y Acad Sci, 2005; 1063:1–12.
38.
RegneryRL, SpruillCL, PlikaytisBD. Genotypic identification of rickettsiae and estimation of intraspecies sequence divergence for portions of two rickettsial genes. J Bacteriol, 1991; 173:1576–1589.
39.
RolainJM, MaurinM, VestrisG, RaoultD. In vitro susceptibilities of 27 rickettsiae to 13 antimicrobials. Antimicrob Agents Chemother, 1998; 42:1537–1541.
40.
RolainJM, StuhlL, MaurinM, RaoultD. Evaluation of antibiotic susceptibilities of three rickettsial species including Rickettsia felis by a quantitative PCR DNA assay. Antimicrob Agents Chemother, 2002; 46:2747–2751.
41.
RouxV, FournierPE, RaoultD. Differentiation of spotted fever group rickettsiae by sequencing and analysis of restriction fragment length polymorphism of PCR-amplified DNA of the gene encoding the protein rOmpA. J Clin Microbiol, 1996; 34:2058–2065.
42.
RouxV, RaoultD. Phylogenetic analysis of members of the genus Rickettsia using the gene encoding the outer-membrane protein rOmpB (ompB). Int J Syst Evol Microbiol, 2000; 50:1449–1455.
43.
RozentalT, EremeevaME, PaddockCD, ZakiSR, et al.Fatal case of Brazilian spotted fever confirmed by immunohistochemical staining and sequencing methods on fixed tissues. Ann NY Acad Sci, 2006; 1078:257–259.
44.
SantibáñezS, PortilloA, PalomarAM, Bell-SakyiL, et al.Isolation and maintenance of Rickettsia raoultii in a Rhipicephalus sanguineus tick cell line. Microbes Infect, 2015; 17:866–869.
45.
SantibáñezS, PortilloA, SantibáñezP, PalomarAM, et al.Usefulness of rickettsial PCR assays for the molecular diagnosis of human rickettsioses. Enferm Infec Microbiol Clin, 2013; 31:283–288.
46.
SekeyovaZ, RouxV, RaoultD. Phylogeny of Rickettsia spp. inferred by comparing sequences of ‘gene D’, which encodes an intracytoplasmic protein. Int J Syst Evol Microbiol, 2001; 51:1353–1360.
47.
SinghS, EldinC, KowalczewskaM, RaoultD. Axenic culture of fastidious and intracellular bacteria. Trends Microbiol, 2013; 21:92–99.
48.
StenosJ, GravesSR, UnsworthNB. A highly sensitive and specific real-time PCR assay for the detection of spotted fever and typhus group rickettsiae. Am J Trop Med Hyg, 2005; 73:1083–1085.
49.
TeysseireN, RaoultD. Comparison of Western immunoblotting and microimmunofluorescence for diagnosis of Mediterranean spotted fever. J Clin Microbiol, 1992; 30:455–460.
50.
UchiyamaT. Growth of typhus group and spotted fever group rickettsiae in insect cells. Ann N Y Acad Sci, 2005; 1063:215–221.
51.
VestrisG, RolainJM, FournierPE, BirgML, et al.Seven years' experience of isolation of Rickettsia spp. from clinical specimens using the shell vial cell culture assay. Ann N Y Acad Sci, 2003; 990:371–374.
52.
WalkerDH, IsmailN. Emerging and re-emerging rickettsioses: Endothelial cell infection and early disease events. Nat Rev Microbiol, 2008; 6:375–386.
53.
WeisburgWG, BarnsSM, PelletierDA, LaneDJ. 16S ribosomal DNA amplification for phylogenetic study. J Bacteriol, 1991; 173:697–703.
54.
WölfelR, EssbauerS, DoblerG. Diagnostics of tick-borne rickettsioses in Germany: A modern concept for a neglected disease. Int J Med Microbiol, 2008; 298:368–374.
55.
WoodsGL, WalkerDH. Detection of infection or infectious agents by use of cytologic and histologic stains. Clin Microbiol Rev, 1996; 9:382–404.
Supplementary Material
Please find the following supplemental material available below.
For Open Access articles published under a Creative Commons License, all supplemental material carries the same license as the article it is associated with.
For non-Open Access articles published, all supplemental material carries a non-exclusive license, and permission requests for re-use of supplemental material or any part of supplemental material shall be sent directly to the copyright owner as specified in the copyright notice associated with the article.
