Volume: 7 Table of Contents: I. N ENG J MED: Ehrlichia ewingii, a Newly Recognized Agent of Human Ehrlichiosis II. J CLIN MICROBIOL: Isolation of DNA after extraction of RNA To detect the presence of Borrelia burgdorferi and expression of host cellular genes from the same tissue sample. III. J CLIN MICROBIOL: Infection of laboratory mice with the human granulocytic ehrlichiosis agent does not induce antibodies to diagnostically significant Borrelia burgdorferi antigens. IV. PEDIATRICS: Characterization of Lyme meningitis and comparison with viral meningitis in children. V. J KOREAN MED SCI: Serologically diagnosed Lyme disease manifesting erythema migrans in Korea. VI. ABOUT THE LYMENET NEWSLETTER Newsletter: *********************************************************************** * The National Lyme Disease Network * * http://www.LymeNet.org/ * * LymeNet Newsletter * *********************************************************************** Volume 7 / Number 07 / 23-JUL-1999 INDEX I. N ENG J MED: Ehrlichia ewingii, a Newly Recognized Agent of Human Ehrlichiosis II. J CLIN MICROBIOL: Isolation of DNA after extraction of RNA To detect the presence of Borrelia burgdorferi and expression of host cellular genes from the same tissue sample. III. J CLIN MICROBIOL: Infection of laboratory mice with the human granulocytic ehrlichiosis agent does not induce antibodies to diagnostically significant Borrelia burgdorferi antigens. IV. PEDIATRICS: Characterization of Lyme meningitis and comparison with viral meningitis in children. V. J KOREAN MED SCI: Serologically diagnosed Lyme disease manifesting erythema migrans in Korea. VI. ABOUT THE LYMENET NEWSLETTER =====*===== I. N ENG J MED: Ehrlichia ewingii, a Newly Recognized Agent of Human Ehrlichiosis ----------------------------------------------------------------- AUTHORS: Buller RS, Arens M, Hmiel SP, Paddock CD, Sumner JW, Rikihisa Y, Unver A, Gaudreault-Keener M, Manian FA, Liddell AM, Schmulewitz N, Storch GA ORGANIZATION: Edward Mallinckrodt Department of Pediatrics and the Department of Medicine, Washington University School of Medicine, St. Louis, MO. REFERENCE: N Engl J Med 1999;341:148-55 ABSTRACT: BACKGROUND: Human ehrlichiosis is a recently recognized tick-borne infection. Four species infect humans: Ehrlichia chaffeensis, E. sennetsu, E. canis, and the agent of human granulocytic ehrlichiosis. METHODS: We tested peripheral-blood leukocytes from 413 patients with possible ehrlichiosis by broad-range and species-specific polymerase- chain-reaction (PCR) assays for ehrlichia. The species present were identified by species-specific PCR assays and nucleotide sequencing of the gene encoding ehrlichia 16S ribosomal RNA. Western blot analysis was used to study serologic responses. RESULTS: In four patients, ehrlichia DNA was detected in leukocytes by a broad-range PCR assay, but not by assays specific for E. chaffeensis or the agent of human granulocytic ehrlichiosis. The nucleotide sequences of these PCR products matched that of E. ewingii, an agent previously reported as a cause of granulocytic ehrlichiosis in dogs. These four patients, all from Missouri, presented between May and August 1996, 1997, or 1998 with fever, headache, and thrombocytopenia, with or without leukopenia. All had been exposed to ticks, and three were receiving immunosuppressive therapy. Serum samples obtained from three of these patients during convalescence contained antibodies that reacted with E. chaffeensis and E. canis antigens in a pattern different from that of humans with E. chaffeensis infection but similar to that of a dog experimentally infected with E. ewingii. Morulae were identified in neutrophils from two patients. All four patients were successfully treated with doxycycline. CONCLUSIONS: These findings provide evidence of E. ewingii infection in humans. The associated disease may be clinically indistinguishable from infection caused by E. chaffeensis or the agent of human granulocytic ehrlichiosis. =====*===== II. J CLIN MICROBIOL: Isolation of DNA after extraction of RNA To detect the presence of Borrelia burgdorferi and expression of host cellular genes from the same tissue sample. ------------------------------------------------------------------- AUTHORS: Amemiya K, Schaefer H, Pachner AR ORGANIZATION: National Institute of Neurological Diseases and Stroke, National Institutes of Health, Bethesda, Maryland 20892, USA. REFERENCE: J Clin Microbiol 1999 Jun;37(6):2087-9 ABSTRACT: We are investigating the neuropathogenesis of Lyme disease caused by Borrelia burgdorferi in a nonhuman primate model. In the past, two separate pieces of tissue had to be used when both analyzing for the presence of the spirochete and examining the host response to infection. We have modified a procedure to purify DNA from the same sample after the extraction of RNA. The remaining material containing the DNA was precipitated, and residual organic reagent was removed prior to deproteinization and extraction of the DNA. This procedure now allows us to both assay for the presence of the Lyme microorganism and analyze the host response in the same tissue preparation. =====*===== III. J CLIN MICROBIOL: Infection of laboratory mice with the human granulocytic ehrlichiosis agent does not induce antibodies to diagnostically significant Borrelia burgdorferi antigens. ------------------------------------------------------------------- AUTHORS: Bunnell JE, Magnarelli LA, Dumler JS ORGANIZATION: Department of Molecular Microbiology and Immunology, The Johns Hopkins University School of Hygiene and Public Health, Baltimore, Maryland 21205, USA. REFERENCE: J Clin Microbiol 1999 Jun;37(6):2077-9 ABSTRACT: Laboratory diagnosis of Borrelia burgdorferi is routinely made by an enzyme-linked immunosorbent assay, with positive results confirmed by Western blot analysis. Concern has been raised that false-positive diagnoses may be made on the basis of serologic cross-reactivity with antibodies directed against other bacterial pathogens, in particular the agent of human granulocytic ehrlichiosis (HGE). The present study made use of a mouse model to ascertain the validity of these concerns. Two different strains of mice were inoculated with the HGE agent and assayed for production of polyclonal and monoclonal antibodies to antigens of both of these bacteria. Infection of mice with the HGE agent does not induce diagnostically significant B. burgdorferi serologic cross-reactions. =====*===== IV. PEDIATRICS: Characterization of Lyme meningitis and comparison with viral meningitis in children. -------------------------------------------------------------------- AUTHORS: Eppes SC, Nelson DK, Lewis LL, Klein JD ORGANIZATION: Division of Infectious Diseases, Alfred I. duPont Hospital for Children, Wilmington, Delaware, USA. [email protected] REFERENCE: Pediatrics 1999 May;103(5 Pt 1):957-60 ABSTRACT: OBJECTIVES: The objectives of this study were to characterize Lyme meningitis (LM) in the pediatric population; to compare LM with viral meningitis (VM) with respect to epidemiology, history and physical examination, and laboratory data; and to provide means of early distinction of Lyme neuroborreliosis from other forms of aseptic meningitis. METHODS: This retrospective analysis involved children admitted to Alfred I. duPont Hospital for Children between 1990 and 1996 whose discharge diagnoses indicated viral or aseptic meningitis or Lyme disease. LM was defined as the presence of cerebrospinal fluid (CSF) pleocytosis with positive Lyme serology and/or erythema migrans. Patients were considered to have VM if they exhibited CSF pleocytosis and had a positive viral culture. Demographic, clinical, and laboratory data were collected for each patient, and patients with LM were compared with age-matched patients with VM. RESULTS: Of 179 patient records, 12 patients with LM and 10 patients with VM (all, >2 years old) were identified by using the above criteria. In comparing LM patients with VM patients, we noted no differences among demographic variables. Children with LM had significantly lower temperatures at the time of presentation. The presence of headache, neck pain, and malaise was similar for the two groups, but the duration of these symptoms was significantly longer among LM patients. Five children with LM had cranial neuropathies. All but 1 LM patient exhibited either papilledema, erythema migrans, or cranial neuropathy. These three findings were absent in the VM group. On CSF analysis, LM patients had fewer white blood cells (mean, 80/mm3 versus 301/mm3) and a significantly greater percentage of mononuclear cells than the VM patients. CONCLUSIONS: In this study, in a Lyme-endemic area, LM was about as common as VM in older children who were hospitalized with aseptic meningitis. Attention to pertinent epidemiologic and historical data, along with physical and CSF findings, allows early differentiation of LM from VM. =====*===== V. J KOREAN MED SCI: Serologically diagnosed Lyme disease manifesting erythema migrans in Korea. ------------------------------------------------------------ AUTHORS: Kim TH, Choi EH, Lee MG, Ahn SK ORGANIZATION: Department of Dermatology, Yonsei University, Wonju College of Medicine, Korea. REFERENCE: J Korean Med Sci 1999 Feb;14(1):85-8 ABSTRACT: Lyme disease is a vector-borne infection, primarily transmitted by Ixodes ticks, and caused by Borrelia burgdorferi. It has a wide distribution in the northern hemisphere. In Korea, however, only one human case has been reported, although B. burgdorferi was isolated from the vector tick I. persulcatus in the region. A 60-year-old male and a 45-year-old female developed the clinical sign of erythema migrans. Each patients were bitten by a tick four weeks and five weeks, respectively, before entering the hospital. On serologic examination, significantly increased IgM and IgG antibody titers to B. burgdorferi were observed in consecutive tests performed at an interval of two weeks. They responded well to treatment with tetracycline. =====*===== VI. 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