2018
DOI: 10.1016/j.bjm.2017.09.004
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Abstract: A modified TaqMan real-time polymerase chain reaction targeting a 138 bp fragment within the lipl32 gene was developed to identify exclusively pathogenic Leptospira spp. in dog urine samples. Thirty-five samples from dogs with suspected clinical leptospirosis and 116 samples from apparently healthy dogs were tested for presence of leptospiral DNA using the TaqMan-based assay. The results were compared with those from a well-established conventional PCR targeting the 16S RNA encoding gene associated with nucleo… Show more

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Cited by 21 publications
(15 citation statements)
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“…isolates were recovered from whole blood (n = 3), urine (n = 7) and abdominal effusion (n = 1) samples. Comparing with previous studies, leptospires were successfully recovered only from urine sample of the diseased dogs [46], shelter and stray dogs [47] and farm dogs [20]. This supports that urine samples are superior samples for leptospiral isolation in dogs regardless of the target population.…”
Section: Discussionsupporting
confidence: 72%
“…isolates were recovered from whole blood (n = 3), urine (n = 7) and abdominal effusion (n = 1) samples. Comparing with previous studies, leptospires were successfully recovered only from urine sample of the diseased dogs [46], shelter and stray dogs [47] and farm dogs [20]. This supports that urine samples are superior samples for leptospiral isolation in dogs regardless of the target population.…”
Section: Discussionsupporting
confidence: 72%
“…The detection of pathogenic Leptospira was performed using a quantitative real-time assay targeting the lipl32 gene. The primers (forward: ; reverse: ), probe ( ) and cycling conditions used had been previously validated to detect pathogenic Leptospira from canine urine samples [ 39 ]. Each reaction had a final volume of 25 μL, with 600 μM of each primer, 250 nM of the probe, 1x TaqMan ® Universal Master Mix II (Thermo Fisher Scientific Inc., Carlsbad, CA, USA), DNase free-water and 5 μL of the extracted DNA.…”
Section: Methodsmentioning
confidence: 99%
“…To confirm the lipl32 qPCR results, all positive samples were subjected to a partial 16S rRNA gene amplification targeting a 331bp fragment [ 41 , 42 ] and were thereafter sequenced to confirm the leptospiral sequence identity. Conventional PCR amplification was carried out as previously described [ 39 ]. Selected samples presenting positive yields were also subjected to a partial secY gene amplification, according to a previous description [ 41 ].…”
Section: Methodsmentioning
confidence: 99%
“…DNA concentration was determined by NanoVue Plus Spectrophotometer (GE Healthcare, USA). The number of Genomic Equivalents (GE), assuming one genome copy per Leptospira, was estimated based on a genomic size of 4.659 Mb (Ren et al 2003, Nascimento et al 2004, Miotto et al 2017.…”
Section: Methodsmentioning
confidence: 99%