Development of SYBR Green real-time PCR for diagnosis of fasciolosis in sheep.
Vet Parasitol
; 283: 109193, 2020 Jul.
Article
en En
| MEDLINE
| ID: mdl-32731054
Fasciolosis is commonly diagnosed by microscopic detection of egg following sedimentation. However, this technique is time-consuming when a large number of samples must be processed and requires sufficient experience. Quantitative real-time PCR based on the detection of liver fluke ribosomal DNA in feces has been introduced, which is more accurate and liable to reflect the presence of flukes in hosts. This study aimed to develop an efficient molecular detection method in laboratory diagnosis. A cross-sectional study of 250 sheep was performed to detect Fasciola hepatica infections using gold standard microscopic detection, conventional PCR and real-time PCR. Both conventional and real-time PCRs targeted the internal transcribed spacer 2 (ITS-2). A composite reference standard(CRS) was used to analyze the sensitivity and specificity of three methods. Furthermore, the minimal amount of plasmid DNA detected by the real-time PCR was 1.67â¯pg plasmid DNA (equivalent to 1.1â¯×â¯106 copies). In conclusion, a highly sensitive and specific method for fasciolosis in sheep was developed.
Palabras clave
Texto completo:
1
Banco de datos:
MEDLINE
Asunto principal:
Quinolinas
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Enfermedades de las Ovejas
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Pruebas Diagnósticas de Rutina
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Diaminas
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Benzotiazoles
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Fascioliasis
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Reacción en Cadena en Tiempo Real de la Polimerasa
Tipo de estudio:
Diagnostic_studies
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Observational_studies
Límite:
Animals
Idioma:
En
Revista:
Vet Parasitol
Año:
2020
Tipo del documento:
Article