Validation of a Real Time PCR for Classical Swine Fever Diagnosis
Natanael Lamas Dias,Antônio Augusto Fonseca Júnior,Anapolino Macedo de Oliveira,Érica Bravo Sales,Bruna R.C. Alves,Fernanda Alves Dorella,Marcelo Fernandes Camargos +6 more
TLDR
RT-qPCR can be used as a complementary diagnostic for CSF and RNA from CSF virus was not detected in samples of swine originated from the Brazilian CSF-free zone.Abstract:
The viral disease classical swine fever (CSF), caused by a Pestivirus, is one of the major causes of economic losses for pig farming. The aim of this work was to validate a RT-qPCR using Taqman for detection of CSF in swine tissues. The parameters for the validation followed the specifications of the Manual of Diagnostic Tests and Vaccines for Terrestrial Animals of the World Organization for Animal Health (OIE) and the guide ABNT NBR ISO/IEC 17025:2005. The analysis of the 5′NTR region of CSF virus was performed in 145 samples from 29 infected pigs and in 240 samples from 80 pigs originated in the Brazilian CSF-free zone. The tissues tested were spleen, kidney, blood, tonsils, and lymph nodes. Sequencing of the positive samples for 5′NTR region was performed to evaluate the specificity of the RT-qPCR. Tests performed for the RT-qPCR validation demonstrated that the PCR assay was efficient in detecting RNA from CSF virus in all materials from different tissues of infected animals. Furthermore, RNA from CSF virus was not detected in samples of swine originated from the Brazilian CSF-free zone. Hence, it is concluded that RT-qPCR can be used as a complementary diagnostic for CSF.read more
Citations
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Simultaneous detection of classical swine fever virus and porcine circovirus 3 by SYBR green I-based duplex real-time fluorescence quantitative PCR.
TL;DR: The developed method was a reliable diagnostic tool to monitor and surveyCSFV, PCV3 and CSFV/PCV3 co-infection in the field and exhibited high repeatability and reproducibility with a low coefficient of variation below 2.0%.
Journal ArticleDOI
Development and validation of rt-qpcr for vesicular stomatitis virus detection (Alagoas vesiculovirus)
Anapolino Macedo de Oliveira,Anapolino Macedo de Oliveira,Antônio Augusto Fonseca Júnior,Marcelo Fernandes Camargos,Lívia Orzil,Mateus Laguardia-Nascimento,Anna Gabriella Guimarães Oliveira,Jacqueline Gomes Rodrigues,Mariana Lázaro Sales,Tatiana Flávia Pinheiro de Oliveira,Cristiano Barros de Melo +10 more
TL;DR: The results showed that RT-qPCRs had excellent and specificity in the detection of RNA of the Alagoas vesiculovirus, indicating that the tests presented excellent repeatability and reproducibility.
Journal ArticleDOI
An outbreak of classical swine fever in pigs in Bangladesh, 2015.
TL;DR: The investigation suggests that CSF is circulating in pigs, posing a risk for communities in Bangladesh that rely on pigs for economic income and dietary protein.
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Validation of a real-time PCR assay for detection of swinepox virus.
Felipe Augusto de Souza,Erlânio Marcelo Dos Santos Júnior,Mateus Laguardia-Nascimento,Tânia R.P. Freitas,Clarissa R. Damaso,Anselmo Vasconcelos Rivetti Júnior,Marcelo Fernandes Camargos,Antônio Augusto Fonseca Júnior +7 more
TL;DR: qPCR for SWPV is a new method with tested variables that allows main sources of error in laboratory diagnosis and viral quantification to be identified and the greatest source of error was variation between analysts rather than different qPCR kits or equipment.
Journal ArticleDOI
RT-qPCR for the diagnosis of the vesiculovirus Cocal virus.
Mariana Lázaro Sales,Mariana Dall’Agnol,Anapolino Macedo de Oliveira,Marcelo Fernandes Camargos,Antônio Augusto Fonseca,Jenner Karlisson Pimenta dos Reis +5 more
TL;DR: RT-qPCR was able to distinguish COCV from other viruses that cause vesicular diseases, an important factor because seroneutralization may produce cross-reactivity between C OCV and vESicular stomatitis Alagoas virus (VSAV).
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