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Medical Microbiology Section Identification of the causative agent of SARS Zusammenfassung SARS (schweres akutes Atemwegssyndrom) verursachte in der ersten Hälfte des Jahres 2003 eine weltweite Epidemie. In Zusammenarbeit mit anderen Europäischen Institutionen gelang der Abteilung für Virologie die Identifizierung des Erregers als ein neues Coronavirus. Auch die ersten RT-PCR Tests für den Erreger wurden von BNI-Wissenschaftlern entwickelt. Summary A severe acute respiratory syndrome (SARS) caused a global epidemic in the first half of 2003. In collaboration with other European insitutions, the Department of Virology identified the causative agent of this novel disease as a new coronavirus. The first diagnostic RT-PCR tests for the new agent were developed by BNITM researchers. Project Description and Results In November 2002 the first case of a new infectious lung disease, termed the severe acute respiratory syndrome (SARS), occurred in Gungdong province of south China. During the subsequent months the epidemic spread to all five continents and eventually came to a halt in July 2003 after strict quarantine measures had been imposed. It claimed the lifes of about 10% of patients, with more than 8000 cases in total. When SARS was imported into Germany in early March 2003, the causative agent was still unknown. SARS-Patients were treated in the University Hospital of Frankfurt and in a pulmonary care facility in Hemer/North Rhine Westphalia. In close collaboration with the Institute of Virology in Frankfurt and other European institutions the Bernhard Nocht Institute identified a novel coronavirus as the causative agent of SARS. A physician who had treated a SARS patient in Singapore was quarantined in the University Hospital of Frankfurt after being disembarked from an intercontinental flight. After a Vero cell culture inoculated with his sputum in the University of Frankfurt showed a cytopathic effect, culture supernatant was transferred to Hamburg for genetic analysis. With an RT-PCR method using 15 different degenerated primer pairs random amplification fragments were obtained from the cell culture supernatant and sequenced. The sequences showed no close similarity to any known DNA sequence, but its translated amino acid code resembled proteins of various animal coronaviruses. The patient meanwhile had made a good clinical recovery and Frankfurt virologists found antibodies in his se- 68 rum that reacted with the infected culture cells. No such antibodies were found in serum taken from the same patient in an earlier phase of the disease. Obviously the cells contained a coronavirus-like agent that caused an antibody response along with clinical recovery. Phylogenetic analysis confirmed a distant but clear relationship of the new agent with known coronaviruses (Figure 1). Figure 1 The PCR Laboratory used the sequence information they had obtained to design several sets of specific polymerase chain reaction primers and established diagnostic tests for the new virus. With the help of a biotech company (Tib-Mobiol, Berlin), RT-PCR primers were made available to laboratories worldwide. Test protocols were distributed via ProMed mail, and positive control material was made available by the Bernhard Nocht Institute. The Institute Pasteur used these tests to examine samples from a cohort of SARS patients shipped to Paris from Vietnam. The test detected the virus in all patients classified as »probable SARS«, in a fraction of patients classified as »suspected SARS«, and in none of the healthy controls (Table 1). Table 1
The same or a highly similar virus was thus present in patients with SARS in two independent local outbreaks (Vietnam and Singapore). When CDC Atlanta made available a different sequence fragment of a virus from another SARS patient, the same fragment was also found in the virus from Frankfurt. The sequence was identical to the base with that from Atlanta. Again, the only link between the patients was the virus. Laboratories in Singapore, Hong Kong, and China were soon able to detect the virus in huge fractions of SARS patients. The PCR Laboratory has also established a real-time RT-PCR assay fully suitable for routine diagnostic application. This test was transformed into the first diagnostic test kit for SARS-Coronavirus by Artus Biotech, a spin-off company of the Bernhard Nocht Institute. It was launced on April 14 th , 2003, one day before the World Health Organization officially declared the new coronavirus to be the cause of SARS. Selected Publications • Drosten C, Gunther S et al.Preiser W, Van Der Werf S, Brodt HR, Becker S, Rabenau H, Panning M, Kolesnikova L, Fouchier RA, Berger A, Burguiere AM, Cinatl J, Eickmann M, Escriou N, Grywna K, Kramme S, Manuguerra JC, Muller S, Rickerts V, Sturmer M, Vieth S, Klenk HD, Osterhaus AD, Schmitz H, Doerr HW. (2003). Identification of a Novel Coronavirus in Patients with Severe Acute Respiratory Syndrome. N Engl J Med 348:1965-74. • Rota PA, Oberste MS, Monroe SS, Nix WA, Campagnoli R, Icenogle JP, Penaranda S, Bankamp B, Maher K, Chen MH, Tong S, Tamin A, Lowe L, Frace M, DeRisi JL, Chen Q, Wang D, Erdman DD, Peret TC, Burns C, Ksiazek TG, Rollin PE, Sanchez A, Liffick S, Holloway B, Limor J, McCaustland K, Olsen-Rassmussen M, Fouchier R, Gunther S, Osterhaus AD, Drosten C, Pallansch MA, Anderson LJ, Bellini WJ. (2003). Characterization of a Novel Coronavirus Associated with Severe Acute Respiratory Syndrome. Science 300:1394-9. • Drosten C, Chiu LL, Panning M, Leong HN, Preiser W, Tam JS, Gunther S, Kramme S, Emmerich P, Ng WL, Schmitz H, Koay ESC (2004). Evaluation of advanced reverse transcription-PCR assays and an alternative PCR target region for detection of severe acute respiratory syndrome-associated coronavirus. J Clin Microbiol, in press. 69 Funding • Bundesministerium für Gesundheit und soziale Sicherung, Grant No. 325-4539-85/3 Cooperating Partners • Prof. Dr. Hans Wilhelm Doerr, University of Frankfurt • Prof. Dr. Sylvie van der Werf, Institut Pasteur, Paris, France • Prof. Dr. Albert Osterhaus, University of Rotterdam, Netherlands Investigators • Christian Drosten • Stephan Günther • Marcus Panning • Stefanie Kramme • Klaus Grywna • Stefanie Müller • Simon Vieth • Herbert Schmitz Medical Microbiology Section
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BERNHARD NOCHT INSTITUTE FOR TROPIC
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Human genetic variants influencing
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Research Funding / Haushaltsmittel
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Introduction The appearance of SARS
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tive project with the Faculty of Me
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since this group headed by Dr. Chri
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Vorwort Das Erscheinen der schweren
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AIDS-Forschung hat eine lange zurü
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Education and Teaching The objectiv
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Education and Teaching Ziel des Kur
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Education and Teaching Institute Fa
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Education and Teaching Lectures and
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Education and Teaching Training for
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Seminar Programme 2002 Prof. Aileen
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Seminar Programme Dr. Stefan Martin
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Symposia and Meetings 2002 Symposia
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Symposia and Meetings 2003 May 13 t
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Staff Activities Staff Activities A
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Staff Activities PD Dr. Joachim Clo
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Staff Activities Editorial Activiti
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Staff Activities Prof. Dr. Rolf Hor
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Staff Activities Prof. Dr. Christia
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Staff Activities Prof. Dr. Egbert T
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Publications Book chapters and Mono
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Publications Herbst R, Ott C, Jacob
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Publications Supali T, Wibowo H, Ru
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Publications Scholze H and Tannich
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Publications Graefe SE, Meyer BS, M
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Publications Schulz A, Mellenthin K
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Appendix 01. Januar 2002 Das BNI wi
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Appendix 01. Oktober 2002 Gemeinsam
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Appendix 01. April 2003 Ergebnis de
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Appendix 01. Juni 2003 Fernsehrepor
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Appendix November 2003 Inauguration
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Appendix red by solar energy could
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Appendix Ansprache von Dr. Thomas F
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Diary 2004/2005 12.-14. November 20
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