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Comparison of conventional PCR, quantitative PCR, bacteriological culture and the Warthin Starry technique to detect Leptospira spp. in kidney and liver samples from naturally infected sheep from Brazil

Fornazari, Felipe ; Costa da Silva, Rodrigo ; Richini-Pereira, Virginia Bodelão ; Beserra, Hugo Enrique Orsini ; Luvizotto, Maria Cecília Rui ; Langoni, Helio

Journal of microbiological methods, 2012-09, Vol.90 (3), p.321-326 [Periódico revisado por pares]

Amsterdam: Elsevier B.V

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Título:
Comparison of conventional PCR, quantitative PCR, bacteriological culture and the Warthin Starry technique to detect Leptospira spp. in kidney and liver samples from naturally infected sheep from Brazil
Autor: Fornazari, Felipe ; Costa da Silva, Rodrigo ; Richini-Pereira, Virginia Bodelão ; Beserra, Hugo Enrique Orsini ; Luvizotto, Maria Cecília Rui ; Langoni, Helio
Assuntos: agglutination tests ; Animals ; Antibodies, Bacterial - blood ; Bacteriological methods and techniques used in bacteriology ; Bacteriology ; Biological and medical sciences ; blood ; Brazil ; Culture Techniques ; Diagnosis ; diagnostic techniques ; Female ; Fundamental and applied biological sciences. Psychology ; Genes, Bacterial ; Kidney - microbiology ; kidneys ; Leptospira ; Leptospira interrogans - genetics ; Leptospira interrogans - growth & development ; Leptospira interrogans - immunology ; Leptospirosis ; Leptospirosis - blood ; Leptospirosis - diagnosis ; Leptospirosis - microbiology ; Leptospirosis - veterinary ; liver ; Liver - microbiology ; Male ; Microbiology ; PCR ; Public health ; quantitative polymerase chain reaction ; Real-Time Polymerase Chain Reaction - standards ; Reference Standards ; Sensitivity and Specificity ; seroprevalence ; serotypes ; Sheep ; Sheep Diseases - blood ; Sheep Diseases - diagnosis ; Sheep Diseases - microbiology ; slaughterhouses
É parte de: Journal of microbiological methods, 2012-09, Vol.90 (3), p.321-326
Notas: http://dx.doi.org/10.1016/j.mimet.2012.06.005
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Descrição: Leptospirosis is an infectious disease of worldwide importance. The development of diagnostic techniques allows sick animals to be identified, reservoirs to be eliminated and the disease prevented and controlled. The present study aimed to compare different techniques for diagnosing leptospirosis in sheep. Samples of kidney, liver and blood were collected from 465 animals that originated from a slaughterhouse. The sera were analyzed by the Microscopic Agglutination Test (MAT), and kidney and liver samples of seropositive animals were analyzed using four techniques: bacteriological culture, the Warthin Starry (WS) technique, conventional PCR (cPCR), and quantitative PCR (qPCR). With the MAT, 21 animals were positive (4.5%) to serovars Hardjo (n=12), Hebdomadis (n=5), Sentot (n=2), Wolfii (n=1) and Shermani (n=1). Titers were 100 (n=10), 200 (n=2), 400 (n=6) and 1600 (n=3). No animal was positive by bacteriological culture; four animals were positive by the WS technique in kidney samples; six animals were positive by cPCR in kidney samples; and 11 animals were positive by qPCR, eight of which in kidney samples and three in liver. The bacterial quantification revealed a median of 4.3bacteria/μL in liver samples and 36.6bacteria/μL in kidney samples. qPCR presented the highest sensitivity among the techniques, followed by cPCR, the WS technique and bacteriological culture. These results indicate that sheep can carry leptospires of the Sejroe serogroup, and demonstrate the efficiency of quantitative PCR to detect Leptospira spp. in tissue samples. ► We compared different techniques to detect Leptospira spp. in kidney and liver samples from naturally infected sheep. ► The Sejroe serogroup was the most prevalent. ► Molecular tests revealed better performance than traditional ones. ► The real time PCR was the most sensitive test. ► Kidney samples presented higher positivity than liver samples.
Editor: Amsterdam: Elsevier B.V
Idioma: Inglês

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Comparison of conventional PCR, quantitative PCR, bacteriological culture and the Warthin Starry technique to detect Leptospira spp. in kidney and liver samples from naturally infected sheep from Brazil

Fornazari, Felipe ; Costa da Silva, Rodrigo ; Richini-Pereira, Virginia Bodelão ; Beserra, Hugo Enrique Orsini ; Luvizotto, Maria Cecília Rui ; Langoni, Helio

Amsterdam: Elsevier B.V

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