Abstract
The aim of the present study was to consider the wide usage of urinary PCR as an increasingly useful tool for an accurate diagnosis of leptospirosis in livestock. A total of 512 adult animals (300 cattle, 138 horses, 59 goats and 15 pigs), from herds/flocks with reproductive problems in Rio de Janeiro, Brazil was studied by serology and urinary PCR. From the 512 serum samples tested, 223 (43.5 %) were seroreactive (cattle: 45.6 %, horses: 41.3 %, goats: 34%and pigs: 60 %). PCR detected leptospiral DNA in 32.4 % (cattle: 21.6 %, horses: 36.2 %, goats: 77.4 % and pigs: 33.3 %. To our knowledge there is no another study including such a large number of samples (512) from different species, providing a comprehensive analysis of the usage of PCR for detecting leptospiral carriers in livestock. Serological and molecular results were discrepant, regardless the titre, what was an expected outcome. Nevertheless, it is impossible to establish agreement between these tests, since the two methodologies are conducted on different samples (MAT - serum; PCR - urine). Additionally, the MAT is an indirect method and PCR is a direct one. In conclusion, we have demonstrated that urinary PCR should be considered and encouraged as an increasingly useful tool for an accurate diagnosis of leptospirosis in livestock.
References
Adler B, de la Peña MA (2010) Leptospira and leptospirosis. Vet Microbiol 140:287–296
Bomfim MR, Barbosa-Stancioli EF, Koury MC (2008) Detection of pathogenic leptospires in urine from naturally infected cattle by nested PCR. Vet J 178:251–256
Desvars A, Cardinale E, Michault A (2011) Animal leptospirosis in small tropical areas. Epidemiol Infect 139:167–188
Fornazari F, da Silva RC, Richini-Pereira VB, Beserra HE, Luvizotto MC, Langoni H (2012) Comparison of conventional PCR, quantitative PCR, bacteriological culture and the Warthin Starry technique to detect Leptospira spp. in kidney and liver samples from naturally infected sheep from Brazil. J Microbiol Methods 90:321–326
Gamage CD, Koizumi N, Perera AK, Muto M, Nwafor-Okoli C, Ranasinghe S, Kularatne SA, Rajapakse RP, Kanda K, Lee RB, Obayashi Y, Ohnishi M, Tamashiro H (2012) Carrier status of leptospirosis among cattle in Sri Lanka: a zoonotic threat to public health. Transbound Emerg Dis. doi:10.1111/tbed.12014
Hamond C, Martins G, Lilenbaum W, Medeiros MA (2012) PCR detection of leptospiral carriers among seronegative horses. Vet Rec 171:105–106
Hernández-Rodríguez P, Díaz CA, Dalmau EA, Quintero GM (2011) A comparison between polymerase chain reaction (PCR) and traditional techniques for the diagnosis of leptospirosis in bovines. J Microbiol Methods 84:1–7
Higino SS, Santos FA, Costa DF, Santos CS, Silva ML, Alves CJ, Azevedo SS (2013) Flock-level risk factors associated with leptospirosis in dairy goats in a semiarid region of Northeastern Brazil. Prev Vet Med 109:158–161
Koizumi N, Muto MM, Akachi S, Okano S, Yamamoto S, Horikawa K, Harada S, Funatsumaru S, Ohnishi M (2013) Molecular and serological investigation of Leptospira and leptospirosis in dogs in Japan. J Med Microbiol 62:630–636
Lilenbaum W, Varges R, Ristow P, Cortez A, Souza SO, Richtzenhain LJ, Vasconcellos SA (2009) Identification of Leptospira spp. carriers among seroreactive goats and sheep by polymerase chain reaction. Res Vet Sci 87:16–19
Limmathurotsakul D, Turner EL, Wuthiekanun V, Thaipadungpanit J, Suputtamongkol Y, Chierakul W, Smythe LD, Day NP, Cooper B, Peacock SJ (2012) Fool’s gold: why imperfect reference tests are undermining the evaluation of novel diagnostics: a reevaluation of 5 diagnostic tests for leptospirosis. Clin Infect Dis 55:322–331
Lucchesi PMA, Arroyo GH, Etcheverría AI, Parma AE, Seijo AC (2004) Recommendations for the detection of Leptospira in urine by PCR. Rev Soc Bras Med Trop 37:131–134
OIE (2012) Manual of diagnostic tests and vaccines for terrestrial animals, 6ªth edn. World Organisation for Animal Health, Paris
Otaka DY, Martins G, Hamond C, Penna B, Medeiros MA, Lilenbaum W (2012) Serology and PCR for bovine leptospirosis: herd and individual approaches. Vet Rec 170:338
Roy S, Biswas D, Vijayachari P, Sugunan AP, Sehgal SC (2005) A clone of Leptospira interrogans sensu stricto is the major cause of leptospirosis in the archipelago of Andaman and Nicobar Islands, India. Lett Appl Microbiol 41:179–185
Stoddard RA, Gee JE, Wilkins PP, McCaustland K, Hoffmaster AR (2009) Detection of pathogenic Leptospira spp. through TaqMan polymerase chain reaction targeting the LipL32 gene. Diagn Microbiol Infect Dis 64:247–255
Subharat S, Wilson PR, Heuer C, Collins-Emerson JM (2011) Evaluation of a SYTO9 real-time polymerase chain reaction assay to detect and identify pathogenic Leptospira species in kidney tissue and urine of New Zealand farmed deer. J Vet Diagn Invest 23:743–752
Acknowledgements
This study was supported by National Counsel of Technological and Scientific Development (CNPq) and Fundação de Amparo à Pesquisa do Estado do Rio de Janeiro (FAPERJ), Brazil. WL is a CNPq fellow.
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Hamond, C., Martins, G., Loureiro, A.P. et al. Urinary PCR as an increasingly useful tool for an accurate diagnosis of leptospirosis in livestock. Vet Res Commun 38, 81–85 (2014). https://doi.org/10.1007/s11259-013-9582-x
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DOI: https://doi.org/10.1007/s11259-013-9582-x