2. César Henríquez 1 Paul Pachas 2 Phillip Lawyer 3 Larry Laughlin 3 Ciro Maguiña 1 1. Instituto de Medicina Tropical Alexander von Humboldt-Universidad Peruana Cayetano Heredia 2. Oficina General de Epidemiologia 3. Uniformed Services University of the Health Sciences 2002
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4. History and Archeology Bartonellosis has been known since Pre-Inca times. Numerous artistic representations in clay “huacos” depict the chronic phase of the disease. Historians and chronists described a disease with warts in Spanish troops when they arrived for the first time in Coaque-Ecuador. For a long time it was thought that the disease was endemic only in Peru and that it had only one phase: “Peruvian wart” or “Verruga peruana”
5. Historical Figures Dr. Alberto L. Barton (1871-1950) Daniel A. Carrión (1858-1885 ) In 1875 an outbreak, characterized by fever and anemia ( Oroya fever ) occurred in the region of construction of the railroad line between Lima and Oroya. In 1885, Daniel A. Carrion , a Peruvian medical student, inoculated himself with material taken from a patient with Peruvian wart . He subsequently acquired Oroya fever and died a month later. Later, Alberto Barton discovered the etiologic agent of “Carrion’s disease”
12. New foci of Carrion’s disease February 2002 New epidemic areas identified. Mortality during the outbreaks is high. No cases of chronic phase (Peruvian wart)in epidemic areas. No animal reservoir identified.
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16. Etiologic agent: Bartonella bacilliformis Gram negative aerobic, pleomorphic, flagellated, motile, coccobacillary, 2-3 m large and 0,2 - 0,5 m wide and facultative intracellular bacterium. For its isolation, special cultures are required containing complemental soy agar, proteases, peptones, some essential amino acids and blood. The optimum growing temperature is 19-29 ºC.
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21. The diagnosis The diagnosis in the acute phase can be done using the thin blood film with Giemsa stain. It is possible to observe the bacillus inside the red blood cells.
22. M: DNA ladder (100 bp). 1: B. bacilliformis DNA from culture extracted by thermal lysis (100°C, 10 min.) using 16S 23S primers (positive control) . 2: Whole blood extraction from an acute phase patient, using 16S 23S primers. 3: Whole blood extraction from an acute phase patient, using primers for Citrate Synthetase gene. 4: B. bacilliformis DNA from a culture extraction using primers for Citrate Synthetase gene . Base pairs 1500 bp 600 bp Molecular technics M 1 2 3 4
23. Lane A: Positive control pool Lane Band C: Bartonella bacilliformis -positive serum taken from a patient in acute phase Lane D: Negative control pool Immunologic technics: Sonicated immunoblot A B C D 20 kDa 18 kDa 17 kDa 14 kDa