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(es)

Fornazari, Felipe ^[1] ; da Silva, Rodrigo Costa ^[1] ; Richini-Pereira, Virginia Bodelao ^[1] ; Orsini Beserra, Hugo Enrique ^[2] ; Rui Luvizotto, Maria Cecilia ^[2] ; Langoni, Helio ^[1]

Afiliação do(s) autor(es)

^[1] Univ Estadual Paulista, UNESP, Fac Med Vet & Zootecnia, Dept Higiene Vet & Saude, Nucleo Pesquisas Zoonose, BR-18618970 Botucatu, SP - Brazil

^[2] Univ Estadual Paulista, Fac Vet Med, UNESP, Dept Clin Ciruigia & Reprod Anim, BR-16050680 Aracatuba, SP - Brazil

• Tipo de documento: Artigo Científico
• Fonte: Journal of Microbiological Methods; v. 90, n. 3, p. 321-326, SEP 2012.
• Citações Web of Science: 26

Resumo

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 43 bacteria/mu L in liver samples and 36.6 bacteria/mu L 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. Published by Elsevier B.V. (AU)

• Processo FAPESP: 09/02393-4 - Pesquisa de leptospiras em ovinos abatidos, sorologicamente positivos para leptospirose: comparação de técnicas e implicações na saúde pública
• Beneficiário: Helio Langoni
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 09/11686-5 - Pesquisa de leptospiras em ovinos abatidos sorologicamente positivos para leptospirose: comparação de técnicas e implicações na saúde pública
• Beneficiário: Felipe Fornazari
• Modalidade de apoio: Bolsas no Brasil - Mestrado