Development of an immunomagnetic antigen capture system for detecting leptospires in bovine urine

Yan, Keding; Ellis, William; Montgomery, J.M.; Taylor, Mark J.; Mackie, D.P.; McDowell, S.W.J.

doi:10.1016/s0034-5288(98)90006-6

Cited by 14 publications

(6 citation statements)

References 32 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Even though nucleotide data do not supply evidence of infecting serovars, [1,2] amplicons containing 16S rRNA gene sequences allow the identification of the infecting species. In the present study three nucleotide sequences showed high sequence homology to L. borgpetersenii ribosomal genes suggesting infection by serovar Hardjo type Hardjobovis, the most common Leptospira found in cattle around the world [18,19]. It is likely that the advent of new generation sequencing procedures will produce inexpensive high quality sequence very useful for epidemiological studies.…”

Section: Discussionmentioning

confidence: 48%

Evaluation of a Polymerase Chain Reaction for the Diagnosis of Leptospirosis in Cattle~!2010-03-10~!2010-08-06~!2010-09-06~!

Baquero¹,

López²,

Mejía³

et al. 2010

TOVSJ

View full text Add to dashboard Cite

Bovine leptospirosis is a highly prevalent infection worldwide causing serious losses in cattle production and serving as a source for human infection. Diagnosis and assessment of prevalence of this infection in bovine herds is difficult due to limitations of current procedures. The present report describes the adaptation of a polymerase chain reaction (PCR) protocol for detection of leptospiral DNA in bovine urine. The amplification products corresponded to a segment of the Leptospira 16S rRNA gene detected using two sets of primers (A/B and C/D). A total of 547 urine samples from Bos taurus (n=327) and Bos indicus (n=220) were collected from animals in Andean and Coastal regions of Ecuador, either by furosemide-induced urination or from bladders at the slaughterhouse. The results of this research showed a PCR positivity of 13.52% using primers A/B. Bos taurus samples obtained by urination and those obtained from bladder showed a significant difference in PCR positivity (P= 0.036). Differentiation of Leptospira species was preformed by DNA sequencing of the amplified products. Three amplicons showed 90 and 98% sequence identity with L. borgpetersenii and 98% identity with L. inadai. The results of this study suggest that PCR could be an excellent approach for epidemiological studies.

show abstract

Section: Discussionmentioning

confidence: 48%

Evaluation of a Polymerase Chain Reaction for the Diagnosis of Leptospirosis in Cattle~!2010-03-10~!2010-08-06~!2010-09-06~!

Baquero¹,

López²,

Mejía³

et al. 2010

TOVSJ

View full text Add to dashboard Cite

show abstract

“…A chemiluminescent immunoassay (612) was applied to human blood and urine (433) but was no more sensitive than earlier ELISA. More recently, immunomagnetic antigen capture was combined with fluoroimmunoassay to detect as few as 10 2 leptospires/ml in urine of cattle infected with serovar hardjo (654). Inhibitory substances have been reported in urine (4,109,654), indicating the need for treatment of urine prior to testing.…”

Section: Antigen Detectionmentioning

confidence: 99%

Leptospirosis

2001

View full text Add to dashboard Cite

Leptospirosis is a worldwide zoonotic infection with a much greater incidence in tropical regions and has now been identified as one of the emerging infectious diseases. The epidemiology of leptospirosis has been modified by changes in animal husbandry, climate, and human behavior. Resurgent interest in leptospirosis has resulted from large outbreaks that have received significant publicity. The development of simpler, rapid assays for diagnosis has been based largely on the recognition that early initiation of antibiotic therapy is important in acute disease but also on the need for assays which can be used more widely. In this review, the complex taxonomy of leptospires, previously based on serology and recently modified by a genotypic classification, is discussed, and the clinical and epidemiological value of molecular diagnosis and typing is also evaluated

show abstract

“…Recent studies report the immunomagnetic separation (IMS) technique prior to PCR assay as an approach to reduce the effect of inhibitory substances present in biological fluids and food samples (2, 3, 4, 6, 7, 9, 14, 15). For diagnosis of leptospirosis, the IMS-PCR approach was reported to detect Leptospira spp.…”

mentioning

confidence: 99%

“…For diagnosis of leptospirosis, the IMS-PCR approach was reported to detect Leptospira spp. in bovine urine (15) and in human biological fluids (6). In order to improve PCR sensitivity and specificity, we developed a novel IMS-PCR approach by using both magnetic beads in house coated with a monoclonal antibody (mAb) and specific PCR primers for pathogenic Leptospira spp.…”

mentioning

confidence: 99%