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Parachlamydiaceae As Rare Agents of Pneumonia

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Parachlamydiaceae As Rare Agents of Pneumonia LETTERS Jorge Hernandez,*† Parachlamydiaceae FAM is 6-carboxyfluorescein and Jonas Bonnedahl,* TAMRA is 6-carboxytetramethylrho- Jonas Waldenström,*§¶ as Rare Agents of damine). Amplification was carried Helena Palmgren,‡ and Björn Pneumonia out on the ABI 7700 sequence detec- Olsen*‡¶ tion system (TaqMan system, Applied *Research Institute for Zoonotic Ecology To the Editor: Members of the Biosystems), by running 45 cycles, and Epidemiology (RIZEE), Färjestaden, Parachlamydiaceae family are emer- Sweden; †Kalmar County Hospital, Kalmar, with annealing temperature of 52°C Sweden; ‡Umeå University, Umeå, ging intracellular bacteria living in and polymerization temperature of Sweden; §Lund University, Lund, Sweden; amoebae (1,2). Serologic studies have 60°C. To prevent carryover, 200 µ M and ¶Ottenby Bird Observatory, suggested that Parachlamydia acan- Degerhamn, Sweden of uracil triphosphate was part of the thamoeba might be an agent of com- master mixture, and uracil-N-glycosy- References munity-acquired pneumonia transmit- lase was used systematically. Para- ted from a water source (3,4). In a sin- chlamydia acanthamoeba strain Hall 1. D’Aoust PY, Busby DG, Ferns L, Goltz J, gle occasion, 16s rRNA of a member McBurney S, Poppe C, et al. Salmonellosis coccus (kindly provided by T.J. of the Parachlamydiaceae family was in songbirds in the Canadian Atlantic Rowbotham) (3) and sterile water provinces during winter-summer 1997– amplified and sequenced from a bron- were used as positive and negative 1998. Can Vet J 2000;41:54–9. choalveolar lavage sample (5). Thus, controls, respectively. In addition, 2. Cízek A, Literák I, Hejlícek K, Treml F, to specify the role played by the Smola J. Salmonella contamination of the PCR was tested on Chlamydophila Parachlamydiaceae as agents of environment and its incidence in wild birds. pneumoniae and Chlamydia psitacci Zentralbl Veterinarmed B 1994;41:320–7. lower respiratory tract infection, we and four strains of Rickettsia. All but 3. Waldenström J, Broman T, Carlsson I, developed a real-time polymerase one (Rickettsia montana) was nega- Hasselquist D, Achterberg RP, Wagenaar chain reaction (PCR) assay and JA, et al. Prevalence of Campylobacter tive, as were 64 sterile water controls. applied it to 1,200 bronchoalveolar jejuni, Campylobacter lari, and Of the 1,200 bronchoalveolar Campylobacter coli in different ecological lavage samples, taken mainly from lavage samples tested, 5 (0.42%) were guilds and taxa of migrating birds. Appl patients with pneumonia of unknown positive. When PCR was repeated for Environ Microbiol 2002;68:5911–7. cause and received in our diagnostic 4. Brooks KA, Sodeman TM. A clinical eval- those five samples, four were negative microbiology laboratory between uation of the API microtube system for for P. acanthamoeba DNA, and only identification of Enterobacteriaceae. Am J 1997 and 2002. one was a true positive, confirmed by Med Technol 1974;40:55–61. DNA extraction was performed by sequencing the product of the addi- 5. Kauffmann F. Das Kauffmann-White- using the MagNA Pure LC instrument Schema. Ergebn. Mikrobiol 1957;30: tional PCR. The sequence shared and the MagNA Pure LC DNA 160–216. 100% DNA homology with P. acan- 6. Aksoycan N, Meco O, Ozsan K, Tekeli ME, Isolation Kit III (Roche Molecular thamoeba strain Hall coccus Saganak I, Ozuygur B. First isolation of a Biochemicals, Mannheim, Germany). (GenBank accession no. AF490592). strain of Salmonella schleissheim in Turkey Real-time PCR was performed by from a patient with enteritis. Mikrobiyol The patient, a 31-year-old man who using TaqMan technology and target- Bul 1983;4:257–8. was HIV-positive, had pneumonia, 7. Hubálek Z, Sixl W, Mikulásková M, Sixl- ing the gene encoding for a nonmito- cough, and no fever. Chest x-ray Vogel B, Thiel W, Halouzka J, et al. chondrial ATP/ADP translocase examination showed an opacity in the Salmonella in gulls and other free-living (GenBank accession no. AF490592). birds in the Czech republic. Cent Eur J right lung and a bilateral infiltrate. This energy parasite gene is present Public Health 1995;3:21–4. Leukocyte count was 5,000/mm3 with 8. Palmgren H, Sellin M, Bergström S, Olsen only in rickettsiae, chlamydiae, and 80 CD4 cells/mm3; microbiologic B. Enteropathogenic bacteria in migrating plant plastids (6). The master mixture investigations (in which the bron- birds arriving in Sweden. Scand J Infect Dis was prepared from the TaqMan 1997;29:565–8. choalveolar lavage was examined for Universal Master Mix kit (Applied 9. Isenberg HD. Interpretation of growth cul- cytomegalovirus, Chlamydophila ture for stool samples. In: Isenberg HD, edi- Biosystems, Foster City, CA), accord- pneumoniae, Legionella pneumo- tor. Essential procedures for clinical micro- ing to the manufacturer’s instructions, phila, Pneumocystis carinii, myco- biology. Washington: American Society for and included 200 nM of each primer Microbiology; 1998. p. 90–4. bacteria, and Toxoplasma gondii) did (Adp81F 5´- TAGTGATCTGC- 10. Prescott JF, Poppe C, Goltz J, Campbell not identify a causal agent. GD. Salmonella typhimurium phage type TACGGGATTT, Adp84R 5´-TTG- We developed a highly sensitive 40 in feeder birds. Vet Rec 1998;142:732. GATTAGGATATTGCAATTT) and PCR, which could amplify as few as 200 nM of the fluorescent labeled 10 bacteria. The assay results in a rel- Address for correspondence: Björn Olsen, probe (6-FAM-5´-AACCTTGTA- Department of Infectious Diseases, Umeå atively high specificity (1,195/ 1,199; GAAGTAACCTGGAA- 99.67%) because it uses a target gene University, SE-901 87 Umeå, Sweden; fax: +46- GAACCAGC-3´-TAMRA, where 6- 90-13 30 06; email: [email protected] found only in Rickettsiae, Emerging Infectious Diseases • Vol. 9, No. 6, June 2003 755 LETTERS Chlamydiae, and plant plastids, and Olivier Castigliola for technical assis- a long time, but their causative agent uses a specific DNA probe. We con- tance. was not detected until 1976 (1). These siderably decreased the risk of hori- viruses of the genus Hantavirus, fam- zontal and vertical contamination of Gilbert Greub,* Pierre Berger,* ily Bunyaviridae, have >20 geno- the PCR reaction by using uracil and Laurent Papazian,† types. Hantaviruses circulate in wild uracil-N-glycosylase and by keeping and Didier Raoult* rodents within natural foci over reaction cups closed since the first *Université de la Méditerranée, Marseille, Euroasia and North and South amplification cycle. France; and †Hôpital Sainte Marguerite, America. They cause asymptomatic More importantly, our study Marseille, France persistent infections in these small showed that Parachlamydia DNA is mammals. Humans may acquire rarely found in bronchoalveolar References infection accidentally from inhalation lavage samples (0.083%). This sug- 1. Greub G, Raoult D. Parachlamydiaceae, of virus-contaminated aerosols of gests that persons are infrequently potential emerging pathogens. Emerg Infect rodent excreta. Hantavirus genotypes exposed to Parachlamydia organisms Dis 2002;8:625–30. may be nonpathogenic for humans or 2. Greub G, Raoult D. Crescent bodies of and, consequently, members of the Parachlamydia acanthamoeba and its life cause serious diseases with high death Parachlamydiaceae seldom cause cycle within Acanthamoeba polyphaga: an rates. In Eurasia, these pathogens pneumonia in humans. In the only electron micrograph study. Appl Environ involve primarily the kidney and positive sample, whether Para- Microbiol 2002;68:3076–84. cause hemorrhagic fever with renal 3. Birtles RJ, Rowbotham TJ, Storey C, chlamydia originated from bacteria in Marrie TJ, Raoult D. Chlamydia-like obli- syndrome; in North and South the oropharynx, from water, or from a gate parasite of free-living amoebae. Lancet America, these pathogens involve pri- colonization of the lower respiratory 1997;349:925–6. marily the lung and cause hantavirus tract was not known; whether they 4. Marrie TJ, Raoult D, La Scola B, Birtles cardiopulmonary syndrome. RJ, de Carolis E. Legionella-like and other caused the patient’s pneumonia is also amoebal pathogens as agents of communi- First reports on the occurrence of not known. That two strains of ty-acquired pneumonia. Emerg Infect Dis hantaviruses in central Europe origi- Parachlamydia found in amoebae 2001;7:1026–9. nated from former Czechoslovakia were recovered from the nasopharynx 5. Corsaro D, Venditti D, Le Faou A, (2,3) and Germany (4) and date back Guglielmetti P, Valassina M. A new of healthy volunteers (7) favors the chlamydia-like 16s rDNA sequence from a to 1984 and 1985, respectively. The first hypothesis. However, that the clinical sample. Microbiology 2001; first cases of hantavirus disease in positive broncholaveolar lavage spec- 147:515–6. humans in the Czech Republic were imen was taken from an HIV-positive 6. Wolf YI, Aravind L, Koonin EV. reported in 1992 (5). This flulike dis- Rickettsiae and chlamydiae evidence of patient with community-acquired horizontal gene tranfer and gene exchange. ease accompanied by microhematuria pneumonia suggests that Parachla- Trends Genet 1999;15:173–5. was recorded in southern Moravia. mydia might occasionally play a path- 7. Amann R, Springer N, Schonhuber W, Another severe imported case was ogenic role in AIDS patients. Ludwig W, Schmid EN, Muller KD, et al. described in a soldier on active mili- Obligate intracellular bacterial parasites of Moreover, any amoebae-associated Acanthamoebae related to Chlamydia spp. tary duty in the Balkans (6). The first bacteria should be considered as a Appl Environ Microbiol
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