Recommendations for the detection of Leptospira in urine by PCR

Lucchesi, Paula María Alejandra; Arroyo, Guillermo H; Etcheverría, Analía Inés; Parma, A; Seijo, Alfredo

doi:10.1590/s0037-86822004000200003

Cited by 42 publications

(40 citation statements)

References 14 publications

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“…In past, many workers have successfully used G1/G2 (Gravekamp et al, 1993, Oliveira et al, 2003, Cheema et al, 2007, Shivaraj et al, 2009, Mineiro et al, 2011, Dehkordi et al, 2011 and LipL32 primers (Bomfim et al, 2008, Meenambigai et al, 2011, Azizi et al, 2012 for diagnostic purpose. Nevertheless, other primers (LipL21, LipL32, LipL41, 16SrRNA) have also been used by various workers (Van Eys et al, 1989, Lucchesi et al, 2004, Lilenbaum et al, 2009, Shivraj et al, 2009, Ambily et al, 2012 for the detection of leptospiral DNA in various biological materials.…”

Section: Resultsmentioning

confidence: 99%

Identification of Pathogenic Leptospira spp. Carrier Goats Using Polymerase Chain Reaction (PCR)

Vihol¹,

Patel²,

Patel³

et al. 2017

Int.J.Curr.Microbiol.App.Sci

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Section: Resultsmentioning

confidence: 99%

Identification of Pathogenic Leptospira spp. Carrier Goats Using Polymerase Chain Reaction (PCR)

Vihol¹,

Patel²,

Patel³

et al. 2017

Int.J.Curr.Microbiol.App.Sci

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“…Additionally, freezing the sample prior to DNA extraction may cause lysis of the leptospires, and the DNA may subsequently be lost with the supernatant after centrifugation (LUCCHESI et al, 2004). Here, the urine samples were processed without freezing and within two hours after collected to prevent the degradation of the nucleic acid of the bacteria that may occur during freezing.…”

Section: Resultsmentioning

confidence: 99%

Serological and molecular findings in diagnosis of leptospirosis serovar hardjo in a dairy bovine herd

Hashimoto¹,

Chideroli

Ribeiro

et al. 2017

SCA

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The cattle are considered hosts of the Hardjo serovar, causing economic damages due to the reproductive failures like abortions and infertility. The serovar Hardjo usually remains in the reproductive tract and also in the renal tubules where it is eliminated intermittently in the urine for months. Placental remnants, the aborted fetus and contaminated urine promote the permanence of this bacterium within the herd for years. Thus, the objective of this study was to monitor for prolonged period, cows naturally infected with Leptospira ssp. through microbiological culture, serological examination and DNA detection of the pathogen in the urine. The dairy herd was composed of 50 breeding cows with a history of abortion and infertility, without leptospirosis vaccine and located in the northern region of Paraná. Blood and urine samples were collected and laboratorial diagnosis were performed five times at intervals of four months. Blood samples were collected from the all 50 animals and the serum was submitted to the microscopic agglutination test (MAT) for the detection of anti-leptospira antibodies. Of the total cows, 20 showed antibody titres ≥ 1: 100 in MAT and urine samples were collected from only those animals with higher titers to perform nested-PCR (n-PCR) and bacterial isolation per culture. In addition, two urine samples from five animals with antibody titers < 1: 100 were collected in MAT for n-PCR. Serovar Hardjo was considered the most frequent during the serological monitoring of the animals evaluated. The n-PCR technique was able to detect leptospiral DNA in the urine of animals with MAT ≥ 1: 100 antibody titers and urine from animals whose titers were < 1: 100. Sequencing of the leptospiral amplicons shared 100% nucleotide sequence identity with the Leptospira interrogans species. Positive n-PCR results from animals with titers of < 1: 100 suggest that the cut-off of MAT is could be not sufficient to detect renal carriers, so it is also important to use n-PCR as an additional diagnostic tool for identify infected animals with Hardjo serovar and whose serology was negative. reprodutivo e também nos túbulos renais onde é eliminado de forma intermitente na urina por meses. Restos placentários, o feto abortado e a urina contaminada favorecem a permanência dessa bactéria dentro do rebanho por anos. Assim, o objetivo deste estudo foi monitorar por período prolongado, vacas naturalmente infectadas por Leptospira ssp. através de cultura microbiológica, exame sorológico e detecção de DNA do patógeno na urina. O rebanho leiteiro estudado era constituído por 50 vacas reprodutoras com histórico de abortos e infertilidade, sem uso de vacina contra a leptospirose e localizada na região Norte do Paraná. As amostras de sangue e urina foram coletadas e submetidas a análises laboratoriais em cinco vezes em intervalos espaçados de quatro meses. As amostras de sangue foram coletadas dos 50 animais e o soro foi submetido ao teste de soroaglutinação microscópica (SAM) para detecção de anticorpos anti-leptospira. Do ...

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“…Para a extração de DNA das amostras de urina foi utilizado um protocolo modificado (Lucchesi et al, 2004). O mesmo foi previamente testado com urina de gato sabidamente negativa e contaminação com quantidades conhecidas de Leptospira spp.…”

Section: Extração De Dnaunclassified

ANTICORPOS ANTI-Leptospira spp. E LEPTOSPIRÚRIA EM GATOS NA REGIÃO METROPOLITANA DE CURITIBA/PR-BRASIL

Cordeiro

Oliveira

Vieira

2017

AVS

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RESUMO:A leptospirose ainda é pouco elucidada em gatos e o papel da espécie na epidemiologia da doença é indeterminado. Os dados sobre a doença clínica nos mesmos são raros, no entanto, a resposta com anticorpos específicos após infecção é comprovada e apresenta valores de prevalência variando de 5,6 a 33%. Apesar da maior resistência da espécie, não se deve ignorar a possibilidade de transmissão, pois é comprovado que após infecção ou doença pode ocorrer eliminação do patógeno na urina. Neste estudo objetivou-se detectar leptospirúria por reação em cadeia da polimerase (PCR) e anticorpos contra L. interrogans com o uso de soroaglutinação microscópica (SAM) em gatos expostos a fatores associados a infecção na região metropolitana de Curitiba/PR. Foram coletadas amostras de sangue e urina de 65 gatos (13 de São José dos Pinhais e 52 de Pinhais), no período de agosto a novembro de 2016. Ao todo, três animais (4,6%) foram soropositivos para o sorovar Pomona, com títulos variando de 200 a 400. Das 65 amostras de urina, uma (1,53%) foi positiva na PCR. Sendo assim, podemos concluir que gatos da região metropolitana de Curitiba/PR expostos a fatores associados a infecção estão sucetíveis a se contaminarem com Leptospira spp. e podem apresentar leptospirúria.Palavras-chave: felinos; leptospirose; saúde pública; sorologia; zoonose ABSTRACT: Leptospirosis is still poorly known in cats and the role of the specie in the disease epidemiology is indeterminate. Clinical disease descriptions are rare, although cats may produce response with specific antibodies after infection. Serological studies have showed a varying prevalence from 6.96 to 26.7%. Despite the specie's larger resistance, the transmission possibility should not be ignored since is confirmed that after infection or disease the pathogen can be eliminated in the urine. The study's objective was to detect leptospiruria by polymerase chain reaction (PCR) and anti-Leptospira spp. Antibodies by microscopic agglutination test (MAT) in cats exposed to infection association factors in the metropolitan zone of Curitiba / PR. Blood and urine samples were collected from 65 cats (13 from São José dos Pinhais and 52 from Pinhais) from august to november 2016(th). Three animals (4.6%) were seropositive for the Pomona serovar, with titles ranging from 200 to 400. Of all the 65 samples collected, one (1.53%) was PCR positive. As a result, we can conclude that cats from the metropolitan area of Curitiba/PR exposed to infection associated factors are susceptible to contamination with Leptospira spp. and may show leptospiruria.

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Recommendations for the detection of Leptospira in urine by PCR

Abstract: In the present study PCR was applied to detect leptospires in human urine. Several

Cited by 42 publications

References 14 publications

Identification of Pathogenic Leptospira spp. Carrier Goats Using Polymerase Chain Reaction (PCR)

Identification of Pathogenic Leptospira spp. Carrier Goats Using Polymerase Chain Reaction (PCR)

Serological and molecular findings in diagnosis of leptospirosis serovar hardjo in a dairy bovine herd

ANTICORPOS ANTI-Leptospira spp. E LEPTOSPIRÚRIA EM GATOS NA REGIÃO METROPOLITANA DE CURITIBA/PR-BRASIL

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