Rickettsia massiliae human isolation.To the Editor: The number of new rickettsial rickettsial /rick·ett·si·al/ (ri-ket´se-al) pertaining to or caused by rickettsiae.
Relating to, or caused by a member of the genus Rickettsia. species that cause diseases in humans is rapidly increasing (1). Moreover, many of the species first described in ticks have been recently shown to be pathogenic. Of the 10 species or subspecies found to be pathogens after 1984, a total of 7 were first isolated from ticks (2). We report the first isolation of Rickettsia rickettsia (rĭkĕt`sēə), any of a group of very small microorganisms, many disease-causing, that live in vertebrates and are transmitted by bloodsucking parasitic arthropods such as fleas, lice (see louse), and ticks. massiliae from a patient. The bacterium was isolated in Sicily in 1985 and identified in 2005.
A 45-year-old man was hospitalized in Palermo, Italy, on June 6, 1985, for fever and a rash. He had been febrile since May 25 and did not respond to antimicrobial drug treatment using cefamezin, a first-generation cephalosporin cephalosporin (sĕf'əlōspôr`ĭn), any of a group of more than 20 antibiotics derived from species of fungi of the genus Cephalosporium and closely related chemically to penicillin. Cephalosporins, e.g. . On examination, he had a necrotic eschar eschar /es·char/ (es´kahr)
1. a slough produced by a thermal burn, by a corrosive application, or by gangrene.
2. tache noire.
n. on his right ankle, a maculopapular rash on his palms and soles (online Appendix Figure 1, available at http://www.cdc. gov/ncidod/EID/vol12no01/05-0850-G1.htm), and slight hepatomegaly hepatomegaly /hep·a·to·meg·a·ly/ (hep?ah-to-meg´ah-le) enlargement of the liver.
The abnormal enlargement of the liver. Also called megalohepatia. . Leukocyte count was normal; he received tetracyclines Tetracyclines Definition
Tetracyclines are medicines that kill certain infection-causing microorganisms.
Tetracyclines are called "broad-spectrum" antibiotics, because they can be used to treat a wide variety of for 13 days and fully recovered. He seroconverted (from 0 to 1:80 between day 11 and day 24) by indirect immunofluorescence to Rickettsia conorii (R. conorii spot, bioMerieux, Marcy l'Etoile, France).
Four milliliters of heparinized blood sampled before treatment were inoculated in a 25-[cm.sup.2] flask containing Vero cells and incubated at 33[degrees]C in a C[O.sub.2] incubator (1). Direct immunofluorescence test on a sample of the patient's serum was positive 7 days later. The strain was stored for 20 years and tested in 2005 at the Unite des Rickettsies for identification, and R. massiliae was identified. DNA DNA: see nucleic acid.
or deoxyribonucleic acid
One of two types of nucleic acid (the other is RNA); a complex organic compound found in all living cells and many viruses. It is the chemical substance of genes. was extracted from the cell culture supernatant and used as template in 2 previously described polymerase chain reaction polymerase chain reaction (pŏl`ĭmərās') (PCR), laboratory process in which a particular DNA segment from a mixture of DNA chains is rapidly replicated, producing a large, readily analyzed sample of a piece of DNA; the process is (PCR PCR polymerase chain reaction.
polymerase chain reaction
Polymerase chain reaction (PCR) ) assays that targeted a portion of the rickettsial ompA gene as well as a portion of the rickettsial gltA gene (3, 4). Amplification products of the expected size were obtained from this extract but from no concurrently processed control materials, including 3 negative controls. DNA sequencing of the positive PCR products gave 100% identity with R. massiliae for ompA (GenBank accession no. RBU RBU Riksförbundet för Rörelsehindrade Barn Och Ungdomar (Swedish National Association for Disabled Children and Young People)
RBU Regional Business Unit
RBU Rukun Budi Utama (The Hague, The Netherlands) 43792) and 99.9% homology for gltA (GenBank accession no. RSU RSU Restricted Stock Unit
RSU Rogers State University (Claremore, Oklahoma)
RSU Rifiuti Solidi Urbani (Italiano)
RSU Rappresentanza Sindacale Unitaria (Italian Group of Unions) 59720).
R. massiliae was first isolated from Rhipicephalus ticks in Marseilles (5). It is transmitted transovarially in Rhipicephalus turanicus (2). R. massiliae is commonly found in Rhipicephalus sanguineus or R. turanicus in France, Greece, Spain (identified as Bar 29) (6), Portugal, Switzerland, Sicily (D. Raoult, unpub. data), Central Africa, and Mali (2). R. massiliae may be commonly associated with these ticks, which are distributed worldwide.
R. massiliae is grouped phylogenically with Rickettsia rhipicephali and Rickettsia aeschlimannii (online Appendix Figure 2, available at http://www.cdc.gov/ncidod/EID/vol1 2no01/05-0850-G2.htm). Bacteria from this group have a natural resistance to rifampin rifampin (rĭfăm`pĭn), antibiotic used in the treatment of tuberculosis. It is also used to eliminate the meningococcus microorganism from carriers and to treat leprosy, or Hansen's disease. that is associated with an rpoB sequence that is different from that of other rickettsiae. This isolate was not tested for antimicrobial drug susceptibly (7). Rifampin resistance leads us to believe that this isolate may cause a Mediterranean spotted fever-like disease that was described in children in Spain (7,8). Serologic findings were recently reported that showed some patients in Barcelona, Spain, with reactions that indicate R. massiliae (B29 strain) rather than R. conorii (6). However, serologic reactions are only presumptive; isolation from a patient is the required to initially describe a new disease (9).
This Sicilian index case shows that R. massiliae is a human pathogen. It contraindicates using rifampin to treat Mediterranean spotted fever in areas where R. massiliae is endemic, as it cannot as yet be differentiated from R. conorii infection. R. massiliae is a new example of a strain identified in ticks for several years before its first isolation from a human patient (10). The longest delay was observed for Rickettsia parkeri, which was isolated from ticks in 1939 but not from a patient until 2004. Many authors labeled R. parkeri a nonpathogenic rickettsia during this time (1). In the present case, the human isolate was obtained before the tick isolate but was not further identified. When this strain was isolated, R. conorii was the sole Rickettsia sp. found in ticks in southern Europe. Moreover, only 1 tickborne pathogenic Rickettsia sp. was believed to circulate in a single area. Since that time, several tickborne rickettsial diseases have been shown to exist in the same area, which prompted us to retrospectively identify this strain. The patient was reexamined in May 2005, after this identification. He is healthy and has no remaining antibodies against Rickettsia spp.
Giustina Vitale, * Serafino Mansueto, * Jean-Marc Rolain, ([dagger]) and Didier Raoult ([dagger])
* Azienda Ospedaliera Universitaria Policlinico "P. Giaccone," Palermo, Italy; and ([dagger]) Universite de la Mediterranee, Marseille, France
(1.) Raoult D, Roux V. Rickettsioses Rickettsioses
Often severe infectious diseases caused by several diverse and specialized bacteria, the rickettsiae and rickettsia-like organisms. The best-known rickettsial diseases infect humans and are usually transmitted by parasitic arthropod vectors. as paradigms of new or emerging infectious diseases. Clin Microbiol Rev. 1997;10: 694-719.
(2.) Matsumoto K, Ogawa M, Brouqui P, Raoult D, Parola P. Transmission of Rickettsia massiliae in the tick, Rhipicephalus turanicus. Med Vet Entomol. 2005;19:263-70.
(3.) Roux V, Fournier PE, Raoult D. Differentiation of spotted fever group rickettsiae by sequencing and analysis of restriction fragment length polymorphism restriction fragment length polymorphism
n. Abbr. RFLP
Intraspecies variations in the length of DNA fragments generated by the action of restriction enzymes and caused by mutations that alter the sites at which these enzymes act, changing of PCR amplified DNA of the gene encoding the protein rOmpA. J Clin Microbiol. 1996;34:2058-65.
(4.) Roux V, Rydkina E, Eremeeva M, Raoult D. Citrate synthase gene comparison, a new tool for phylogenetic analysis, and its application for the rickettsiae. Int J Syst Bacteriol. 1997;47:252-61.
(5.) Beati L, Raoult L. Rickettsia massiliae sp.nov., a new spotted fever group rickettsia, Int J Syst Bacteriol. 1993;43:839-40.
(6.) Cardenosa N, Segura F, Raoult D. Serosurvey among Mediterranean spotted fever patients of a new spotted fever group rickettsial strain (Bar29). Eur J Epidemiol. 2003;18:351-6.
(7.) Drancourt M, Raoult D. Characterization of mutations in the rpoB gene in naturally rifampin-resistant Rickettsia species. Antimicrob Agents Chemother. 1999;43:2400-3.
(8.) Bella F, Espejo-Arenas E, Uriz S, Serrano JA, Alegre MD, Tort J. Randomized ran·dom·ize
tr.v. ran·dom·ized, ran·dom·iz·ing, ran·dom·iz·es
To make random in arrangement, especially in order to control the variables in an experiment. trial of five-day rifampin versus one-day doxycycline doxycycline /doxy·cy·cline/ (dok?se-si´klen) a semisynthetic broad-spectrum tetracycline antibiotic, active against a wide range of gram-positive and gram-negative organisms; used also as d. calcium and d. hyclate. therapy for Mediterranean spotted fever. J Infect Dis. 1991;164:433-4.
(9.) Parola P, Paddock CD, Raoult D. Tickborne rickettsioses around the world: emerging diseases challenging old concepts. Clin Microbiol Rev. 2005;18:719-56.
(10.) Paddock CD, Sumner JW, Comer JA, Zaki SR, Goldsmith CS, Goddard J, et al. Rickettsia parkeri: a newly recognized cause of spotted lever rickettsiosis rickettsiosis /rick·ett·si·o·sis/ (ri-ket?se-o´sis) infection with rickettsiae.
Infection with Rickettsia bacteria. in the United States. Clin Infect Dis. 2004;38:805-11.
Address for correspondence: Didier Raoult, Unite des Rickettsies, CNRS CNRS Centre National de la Recherche Scientifique (National Center for Scientific Research, France)
CNRS Centro Nacional de Referencia Para El Sida (Argentinean National Reference Center for Aids) UMR 6020, Faculte de Medecine, Universite de la Mediterrande, 27 Bd Jean Moulin, 13385 Marseille CEDEX 05, France; fax: 33-4-91-38-77-72; email: didier.raoult@medecine. univ-mrs.fr
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