Detection of Leptospira in urine of apparently healthy dogs by quantitative polymerase chain reaction in Haryana, India

Rohilla, Preeti; Khurana, Rajesh; Kumar, Aman; Batra, Kanisht; Gupta, Renu

doi:10.14202/vetworld.2020.2411-2415

Cited by 7 publications

(3 citation statements)

References 29 publications

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“…Although several studies have demonstrated high specificity and sensitivity for real-time PCR and found it to be a rapid method for diagnosing acute leptospirosis (Stoddard et al, 2009), in the present study, specificity was high, but sensitivity low. Based on previous research, this can be explained by intermittent urinal Leptospira shedding, prior use of antibiotics before analysis, freezing of samples and the pH of urine, which can influence and reduce test sensitivity (Fraune et al, 2013;Rohilla et al, 2020;Schuller et al, 2015). Troìa et al (2018) also observed low diagnostic sensitivity for qPCR, with sensitivity and specificity of 13.5 and 92.0 %, respectively, similar to our findings.…”

Section: Resultssupporting

confidence: 87%

Alterações clínico-patológicas, diagnóstico sorológico e molecular em cães com suspeita de leptospirose

et al. 2023

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A leptospirose é uma zoonose de importância mundial que causa mortes tanto em humanos, como em animais, especialmente em cães não vacinados. A infecção em cães ocorre pelo contato com urina ou água contaminada pela bactéria espiroqueta patogênica do gênero Leptospira. Objetivou-se avaliar dois testes diagnósticos na caracterização de cães com leptospirose, e associar com as principais alterações clínicos-patológicas de cães suspeitos. Foram atendidos no Hospital Veterinário da Universidade Federal de Uberlândia no período de setembro de 2019 a setembro de 2020, 24 cães com suspeita clínica de leptospirose canina. Exames complementares como hemograma completo, dosagens séricas de creatinina, ureia, ALT (alanina aminotransferase), FA (fosfatase alcalina) foram solicitados. Para o diagnóstico confirmatório de leptospirose foi realizado teste sorológico pelo teste de aglutinação microscópica (MAT) e também teste molecular pela reação em cadeia da polimerase em tempo real (qPCR). Dos 24 casos de cães suspeitos de leptospirose, seis (25%) foram positivos no MAT, e três (12,5%) no qPCR, sendo 1 (um) (4,17%) positivo nos dois testes diagnósticos avaliados. Quando se associou MAT + qPCR a positividade foi para 8/24 (33,3%). Os sorogrupos reativos predominantes foram Icterohaemorrhagiae (33,33%) e Djasiman (33,33%), seguido de Ballum (16,60%). A sensibilidade e especificidade da qPCR em relação teste padrão ouro (MAT) foi 16,7 % e 88,9%, respectivamente. Houve alterações nos níveis séricos de creatinina, ureia e FA no grupo positivo aos testes diagnósticos (MAT/qPCR), porém somente ureia e FA apresentaram valores elevados no grupo negativo. Quando se comparou os parâmetros bioquímicos no grupo positivo aos testes diagnósticos (MAT/qPCR), e negativo, não se observou nenhuma diferença estatística significativa entre os elementos avaliados (p>0,05). Conclui-se para o diagnóstico de leptospirose canina aguda, o médico veterinário deve associar os achados clínicos-patológicos aos resultados do MAT de forma pareada e o qPCR de urina.

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Section: Resultssupporting

confidence: 87%

Alterações clínico-patológicas, diagnóstico sorológico e molecular em cães com suspeita de leptospirose

et al. 2023

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“…The lateral flow immunochromatographic kit (LFK) is a simple, rapid test that can be carried out in the field. Now the LFK technology is widely used for the determination of bacterial pathogens in clinical samples of different diseases, such as S. aureus in clinical samples [ 8 ], S. aureus enterotoxin A in milk [ 9 ], Vibrio cholera [ 10 ], Vibrio harveyi [ 11 ], Yersinia pestis in humans [ 12 ], Leptospira in urine [ 13 ], Helicobacter pylori in stool [ 14 ], Streptococcus suis serotype 2 [ 15 ], Salmonella enterica subsp. Enterica serovar Typhimurium ( S. typhimurium ) [ 16 , 17 ], Candida albicans [ 18 ], and COVID-19 coronavirus [ 19 ].…”

Section: Introductionmentioning

confidence: 99%

Development of a lateral flow device for rapid simultaneous multiple detections of some common bacterial causes of bovine mastitis

Sayed¹,

Soliman²,

Elsaady³

2023

J Adv Vet Anim Res

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Objective: This work was conducted for the development of a 5-combi lateral flow immunochro¬matographic kit (LFK) for rapid and simultaneous identification of the common bacterial causes of bovine mastitis. The following pathogens are the identification targets of this kit: Escherichia coli, Staphylococcus aureus, Klebsiella pneumoniae, Streptococcus agalactiae, and Streptococcus pyogenes in milk samples from suspected bovine mastitis cases. The conventional microbiological identification of these agents is not only time-consuming and requires a fully equipped laboratory but also requires experienced personnel. Materials and Methods: Rabbit polyclonal antibodies (PAbs) specific to the antigenic components of the selected pathogens were prepared, and the pathogen-specific IgG was separated, purified, and conjugated with nanogold that was laid on the conjugate pad. Guinea pig PAbs specific to the microbial antigens of the selected pathogens were prepared, and their IgG content was separated, purified, and used as a capture antibody in the test (T) line on the nitrocellulose (NC) strips. Goat anti-rabbit IgG antibodies were used to capture the rabbit antibodies in the control (C) line of NC strips. The kit was held in a device comprising five strip-holding channels for the above-mentioned five bacterial species antigens. The developed LFK was evaluated, and its sensitivity and specificity were determined. Results: The developed kits were applied for the examination of bovine milk samples from suspected mastitis cases, and the average sensitivity, specificity, and accuracy of 5-combi LFK for the detection of the five selected bacterial species compared to bacteriological examination (gold standard test) were 93.90%, 80.83%, and 90.53%, respectively. The minimal microbial count that gave positive results using the developed LFK was 103 colony forming unit/ml. Treatment of the milk samples with an application buffer and its pre-incubation in trypticase soy broth for 6 h at 37°C before testing significantly increased the sensitivity of the prepared LFK. The developed kit proved simple and convenient, and the results could be obtained in less than 10 min.

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“…7,8 Rats, canines, buffaloes, and cattle are the hosts for leptospirosis. 9 Humans or animals are infected accidentally by direct or indirect contact with leptospira infected animals or contaminated water or urine. 10,11 Infection is also possible by other methods, like handling clinically infected animal tissues, inhalation of droplets of urine, 12 and consuming contaminated food and water.…”

Section: Introductionmentioning

confidence: 99%

Epidemiological Status of Leptospirosis in India

Choudhary,

Kumar

et al. 2023

J. Pure Appl. Microbiol.

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A corkscrew-shaped spirochaete named Leptospira interrogans causes the infectious disease called leptospirosis. Leptospirosis, a growing public health concern worldwide, from subclinical infections to potentially lethal pulmonary hemorrhage. The disease caused by the pathogenic Leptospira, poses a threat to both humans and animals, and its transmission occurs through contact with infected animals, contaminated water, and soil. The prevalence of leptospirosis is influenced by various factors, including climate, urban development, and animal-rearing practices. It can manifest with severe symptoms in humans, making early diagnosis crucial. Diagnostic methods like microscopic agglutination test (MAT) and enzyme-linked immunoassay (ELISA) are widely used for the screening of leptospira infection. Molecular technique like PCR and qPCR offering higher sensitivity and rapidity. This paper meta-analyses the incidence of leptospirosis in various animals based on the literature published from 2005 to 2023 and provides prevalence of the disease in various animal including humans. Results suggest the significance prevalence of the disease in humans and various animal species, namely buffalo, rodents, and dogs. Coastal regions in India were particularly vulnerable to the disease. Efforts to control leptospirosis include surveillance programs and public health initiatives. Understanding the epidemiology and prevalence of leptospirosis, as highlighted in this paper, is essential for implementing effective preventive measures. Finally, a continued research, diagnostic advancements, public awareness campaigns and addressing research gaps in epidemiology of the disease are critical in mitigating the impact on human and animal health. This review provides important data for public health authorities, veterinarians, scientists and for the public, in general.

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Detection of Leptospira in urine of apparently healthy dogs by quantitative polymerase chain reaction in Haryana, India

Cited by 7 publications

References 29 publications

Alterações clínico-patológicas, diagnóstico sorológico e molecular em cães com suspeita de leptospirose

Alterações clínico-patológicas, diagnóstico sorológico e molecular em cães com suspeita de leptospirose

Development of a lateral flow device for rapid simultaneous multiple detections of some common bacterial causes of bovine mastitis

Epidemiological Status of Leptospirosis in India

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