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

Abstract

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.

Polymerase Chain Reaction in the Diagnosis of Spontaneous Leptospirosis in Bovines

“…The PCR positive samples also showed positivity in microscopic agglutination test (MAT) with L. interrogans serovar Hardjo, Canicola, Hebdomadis, Icterohaemorrhagiae, Pomona, Autumnalis, Australis, Pyrogenes; L. kirschneri serovar Grippotyphosa and L. borgpetersenii serovar Javanica (Patel et al, 2014). Further the amplicons size of G1/G2 and LipL32 primers was 285bp and 756bp, respectively and supported the earlier findings (Meenambigai et al, 2011;Gravekamp et al, 1993;Bomfim et al, 2007;Baquero et al, 2010).…”

Identification of cattle, buffaloes and rodents as reservoir animals of Leptospira in the District of Gampaha, Sri Lanka

“…Samples were immediately centrifuged at ~5000× g for 15 min, the pellets were resuspended in 1 ml of 1 × phosphate buffered saline (PBS) [137 mM NaCl, 2.7 mM KCl, 4.3 mM Na 2 HPO 4 , 1.4 mM KH 2 PO 4 (pH 7)], transferred into a 1.5 ml microcentrifuge tube and centrifuged at ~8000× g for 5 min. The supernatant was discarded and pellets resuspended in 1× PBS and stored at −20.0 °C until used for testing [16]. …”

Detection of Brucella sp. and Leptospira sp. in dogs using conventional polymerase chain reaction

“…Sensitivity and specificity of the method are so high, that they are often used to compare other methods such as isolation and culture. PCR technique has been increasingly used for the detection of Brucella and Leptospira in clinical and research applications (2,8). This work demonstrated the presence of DNA of the two studied bacteria, and that means a subclinical acute infection.…”

“…Direct bacteriological isolations can be also conducted, but they are difficult, time consuming and risky. In order to improve the direct diagnosis and minimise the aforementioned problems, molecular diagnosis based on polymerase chain reaction (PCR) has been successfully used for the detection of Leptospira and Brucella in clinical samples (2,14,17).…”