Diagnosis of canine leptospirosis using an immunomagnetic separation-PCR method

Monte, Leonardo Garcia; Jorge, Sérgio; Luiz, João; Sinnott, Francine Alves; Seixas, Kömmling Seixas; Aleixo, José Antônio Guimarães; Samartino, Luis; Conceição, Fabrício Rochedo; Hartleben, Cláudia Pinho

doi:10.1590/s1517-83822012000200023

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“…The rrs conventional PCR was less sensitive in whole blood and urine samples than the lipL32 real-time PCR. However, the rrs conventional PCR showed values higher than those reported in human urine (10 4 bacteria/mL of urine) and similar to those obtained by other authors in canine urine after using a previous stage of immunomagnetic separation to increase sensitivity or a conventional PCR assay targeted hap1 gene (Lucchesi et al 2004, Branger et al 2005, Monte et al 2012.…”

Section: Discussionsupporting

confidence: 88%

“…Conventional or real-time PCR assays previously studied to detect Leptospira spp. target genes common to all Leptospira spp., including the rrs, gyrB, flaB and secY genes, or pathogen-specific genes, including the lipL32, ligA and ligB genes (Mérien et al 1992, Levett et al 2005, Palaniappan et al 2005, Slack et al 2006, Ahmed et al 2009, Rojas et al 2010, Monte et al 2012, Koizumi et al 2013, Xu et al 2014. The advantage of a conventional PCR assay based on the detection of the rrs gene is its low cost, which allows using it in routine clinical diagnosis.…”

Section: Introductionmentioning

confidence: 99%

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Diagnosis of canine leptospirosis: evaluation of two PCR assays in comparison with the microagglutination test

et al. 2019

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Canine leptospirosis is definitely diagnosed by demonstrating seroconversion in paired serum samples from the acute and convalescent period by the microagglutination test (MAT). However, the application of a polymerase chain reaction (PCR) assay can provide earlier confirmation of suspected cases. The objective of this study was to evaluate two PCR assays used in diagnosis of human leptospirosis (lipL32 real-time PCR and rrs conventional PCR) in cultured microorganisms and experimentally contaminated samples (whole blood, serum, urine), and investigate their applicability in clinical samples from dogs with presumptive diagnosis of leptospirosis by using the MAT as a reference. The analytical sensitivity of the lipL32 real-time PCR was 1 genome equivalent per reaction, whereas that for the rrs conventional PCR was 10 genome equivalents per reaction. Both assays amplified the pathogenic strains but were negative when evaluating the DNA of other microorganisms that may be present in clinical samples. The lipL32 real-time PCR detected 100 bacteria/mL in whole blood samples, 1000 bacteria/mL in serum samples and 10 bacteria/mL in urine samples, whereas the rrs conventional PCR detected 1000 bacteria/mL in whole blood and serum samples and 100 bacteria/mL in urine samples. Seven out of the 51 samples from dogs with presumptive diagnosis of leptospirosis were considered as confirmed cases. ThelipL32 real-time PCR detected positive results in six of the seven confirmed cases, whereas the rrs conventional PCR detected four. The PCR assays evaluated proved to be useful diagnostic tools in the confirmation of canine leptospirosis when used together with the MAT.

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Section: Discussionsupporting

confidence: 88%