2013, Number 2
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Rev Cubana Med Trop 2013; 65 (2)
Detection of leptospiral DNA in fresh tissues from dead people in Cuba, 2008-2011
Rodríguez OY, Rodríguez GI, Zamora MY, Rodríguez SJE, Valdés LY, Echevarria PE, Obregón FAM, Verdasquera CD, Fernández MC
Language: Spanish
References: 26
Page: 211-222
PDF size: 161.85 Kb.
ABSTRACT
Introduction: leptospirosis is an important re-emerging zoonosis worldwide, with
an associated mortality ranging 15-25 %. Post-mortem diagnosis of the disease is
painstaking. Different molecular methods for diagnosis of leptospirosis have been
implemented. In Cuba, the National Reference Laboratory for Letospires of "Pedro
Kourí" Institute of Tropical Medicine (LNRL-IPK) performs laboratory diagnosis of
fatal cases suspected of leptospirosis.
Objective: to diagnose leptospirosis in fresh tissue samples from dead people by
using a molecular method.
Methods: of 50 deceased individuals- 3 in 2008, 7 in 2009, 15 in 2010 and 25 of
2011- from different Cuban regions, 171 samples of fresh tissues of different
organs were studied. A singleplex polymerase chain reaction that amplifies
lipl32
gene fragment was used. Additionally, a serological diagnosis of leptospirosis was
performed in 9 cases.
Results: the laboratory confirmed 32 % of cases as positive (16/50). Of these,
2 (66.7 %) in 2008, 2 (28.6 %) in 2009, 1 (6.7 %) in 2010 and 11 (44 %) in 2011.
The positivity rate of the different sample was 25 % in the lung (12/48), 25.6 % in
the liver (11/43), 22.5 % in the kidneys (9/40), 6.3 % in the brain (1/16), 7.1 % in
the heart (1/14), 12.5 % in the spleen (1/8) and 50 % in the muscles (1/2). Of the
nine cases that underwent serology, three were positive by PCR and negative by
serological testing, whereas one case was found positive in the serology but
negative in the PCR. The overall agreement coefficient between serology and PCR
was 55.5 % (5/9).
Conclusions: polymerase chain reaction, as a molecular tool for diagnosis of
leptospirosis at the LNRL-IPK, has allowed making post-mortem confirmation of
that disease in Cuba. It is recommended to continue shipping serum samples from
fatal cases to the LNRL-IPK for confirmation, which will allow a better interpretation
of the results by using serology.
REFERENCES
Berdasquera D, Barreras BA, Barroso J, Pérez A, Fernández C, Ortega LM, et al. Mortalidad por leptospirosis humana. Ciudad de La Habana, 2005-2006. Rev Panam Infectol. 2009;11(4):19-26.
Ko AI, Reis MG, Dourado CMR, Johnson WD, Riley LW, et al. Urban epidemic of severe leptospirosis in Brasil. Lancet. 1999;354:820-5.
Roca B. Leptospirosis. Rev Med Univ Navarra. 2006;50(2):3-6 [citado 22 Jul 2012]. Disponible en: http://www.ncbi.nlm.nih.gov/pubmed/16999233
Verdasquera D, Barroso J, Barreras BA, Pérez A, Pérez K, Obregón AM, et al. Factores asociados a la morbilidad por Leptospirosis humana. Ciudad de La Habana, 2005-2006. Rev Panam Infectol. 2010;12(1):8-16.
Verdasquera D, Ortega LM, Fernández C, Obregón AM, Rodríguez I, Miyar R. Enfrentamiento a brotes epidémicos de leptospirosis humana. Rev Panam Infectol. 2011;13(1):28-35.
Verdasquera D, Galí L, Sánchez L, Alpízar D, Vega B, Calas V. Variación de la morbilidad y mortalidad por leptospirosis humana. Cuba, 1998-2007. Rev Panam Infectol. 2010;12(4):22-30.
Obregón AM, Fernández C, Rodríguez I, Balbis Y, Martínez B, Rodríguez J. Sistema de aglutinación con látex para el diagnóstico rápido de la leptospirosis en Cuba. Rev Panam Salud Publica. 2004;16(4):259-65.
Levett P. Leptospirosis. Clin Microbiol Rev. 2001;14(2):296-320.
Dassanayake DL, Wimalaratna H, Agampodi SB, Liyanapathirana VC, Piyarathna TA, Goonapienuwala BL. Evaluation of surveillance case definition in the diagnosis of leptospirosis, using the Microscopic Agglutination Test: a validation study. BMC Infect Dis. 2009;9:48.
Rodríguez I, Fernández C, Obregón AM, Zamora Y, Rodríguez JE, Rodríguez NM, et al. Confirmación microbiológica de 2 brotes emergentes de leptospirosis humana en Cuba. Rev Cubana Med Trop. 2007;59(1):19-23.
Fernández C, Obregón AM, Rodríguez I, Rodríguez Y, Mondeja BA, Noda AA, et al. Manual de operaciones y procederes para el diagnóstico de la leptospirosis humana. La Habana/ Panamá: Ediciones DAMPSA; 2012. p. 24,25,32,33,59,61, 64, 65.
Brown PD, Gravekamp C, Carrington DG, van de Kemp H, Hartskeerl RA, Edwards CN, et al. Evaluation of the polymerase chain reaction for early diagnosis of leptospirosis. J Med Microbiol. 2006;43(3):110-14.
Adler B, de la Peña-Moctezuma A. Leptospira y leptospirosis. Vet Microbiol. 2010;140:287-96.
Levett P, Morey RE, Galloway RL, Turner DE, Steigerwalt AG, Mayer LW. Detection of pathogenic leptospires by real-time quantitative PCR. J Med Microbiol. 2005;54:45-9.
WHO-ILS. Human leptospirosis: guidance for diagnosis, surveillance and control. Malta: WHO; 2003. p. 5-7, 43, 63, 64, 77.
Verdasquera D, Ortega LM. Leptospirosis humana: pautas para su prevención y control. Ediciones DAMPSA: La Habana/Panamá; 2012. p. 49-57, 63.
Bacich DJ, Sobek KM, Cummings JL, Atwood AA, O´Keefe D. False negative results from using common PCR reagents. BMC Research Notes. 2011;4:457.
Daher E, Trevisan DM, Borges M, Abdulkader RCRM. Risk factors for death and changing patterns in leptospirosis acute renal failure. Am J Trop Med Hyg. 1999;61(4):630-4.
Gouveia E, Metcalfe J, de Carvalho AL, Aires TS, Villasboas-Bisneto JC, Queirroz A, et al. Leptospirosis-associated severe pulmonary hemorrhagic syndrome, Salvador, Brazil. Emerg Infec Dis. 2008;14(3):505-8.
Rodríguez I, Obregón AM, Rodríguez JE, Fernández C, Arzola A, Victoria B. Caracterización serológica de cepas aisladas de pacientes con leptospirosis humana en Cuba. Rev Cubana Hig Epidemiol. 2002;40(1):11-5.
Coyne VE, James MD, Reid SJ, Rybicki EP. Molecular Biology Techniques Manual. 3rd ed. University of Cape Town; 2003 [citado 22 Jul 2012]. Disponible en: http://www.mcb.uct.ac.za/manual//MolBiolManual.htm
Cullen PA, Xu X, Matsunaga J, Sanchez Y, Ko AI, Haake DA. Surfaceome of Leptospira spp. Infect Immun. 2005;73:4853-63.
Gravekamp C, van de Kemp H, Franzen M, Carrington D, Schoone GJ, van Eys G, et al. Detection of seven species of pathogenic leptospires by PCR using two sets of primers. J Gen Microbiol. 1993;139:1691-700.
Haake DA, Chao G, Zuerner RL, Barnett JK, Barnett D, Mazel M, et al. The leptospiral major outer membrane protein LipL32 is a lipoprotein expressed during mammalian infection. Infect Immun. 2000;68(4):2276-85.
Brown PD, Carrington DG, Gravekamp C, van de Kemp H, Edwards CN, Jones SR, et al. Direct detection of leptospiral material in human postmortem samples. Research Microbiology. 2003;154:581-6.
de Abreu C, Teixeira VL, Caló E, Spinosa C, Arroyo MC, da Silva MV, et. al. Polymerase chain reaction in comparison with serological tests for early diagnosis of human leptospirosis. Trop Med Int Health. 2006;2(2):1699-707.