Leptospira DNA PCR - Quantitative Test Cost & Procedure

Abstract

Leptospirosis is a bacterial zoonotic disease brought on by pathogenic Leptospira species and is preventable by vaccination. By administering virulent serovars of Leptospira to control and vaccination dogs, followed by the detection of Leptospira in blood and urine, the effectiveness of canine Leptospira vaccines is evaluated. Utilizing species-distinct  real-time qPCR targeting the lipL32 gene, we approximated the consequences of urine and blood specimens from clinical examinations to those acquired utilizing the consideration culture technique.

Negative results for nonpathogenic Leptospira and for a number of canine viruses, germs, and parasites served as proof of the qPCR assay's specificity. Utilizing McNemar's examination, kappa coefficient , and portion of consensus examinations, the consequences of the two strategies were approximated. There were no false-negative conclusions with the qPCR assay, and the outcomes for the portion of favorable and unfavorable dogs were analogous. There was outstanding consensus between the outcomes of the qPCR and the culture technique for both blood and urine .

Introduction

Leptospira, a pathogenic spirochaete bacteria, is the source of canine leptospirosis, an ailment that affects both domestic and wild dogs. Leptospira infection in dogs can cause asymptomatic to severe, potentially deadly disease with multiorgan dysfunction in the clinical symptoms . One of the most pervasive zoonotic illnesses, leptospirosis affects both humans and animals. Direct contact with diseased animal urine can cause infection, as can being exposed to water or soil that has been polluted by rodent and other wild animal pee .It is advised to vaccinate dogs who may be exposed to pathogenic Leptospira strains in order to prevent them from illness and lower the danger of zoonotic spread by lowering the shedding of virulent Leptospira.

The most prevalent leptospirosis protein, LipL32, a lipoprotein with surface exposure from the outer membrane that is also referred to as Hap1, is only present in pathogenic leptospires. This very immunogenic protein is expressed in vivo and is largely conserved among pathogenic Leptospira strains. LipL32-specific antibodies have been demonstrated to be highly specific, with no detectable cross-reactivity to unrelated antigens, and to be present at all illness phases .

The effectiveness of veterinary C vaccinations is evaluated during clinical development by their capacity to shield dogs against exposure to particular pathogenic Leptospira serovars

According to the European Pharmacopoeia, vaccination effectiveness must be evaluated using protection against challenge with a pathogenic strain in order to determine the timing of immunity (OOI) and the length of immunity (DOI). A reference culture approach is used to test blood, urine, and kidney tissue samples from challenged dogs for the presence of Leptospira for up to 6 weeks following the challenge in order to evaluate protection using clinical parameters.

Since Leptospira growth is slow and can take several weeks, this reference culture method is time-consuming and labor-intensive. Additionally, the cultures must be regularly checked under a light microscope for live leptospires. The test samples' danger of contamination and the possible risk of exposure for technicians both rise as a result of the protracted multistep process for culture and reading.