6 results on '"Phillip Ormeño-Vásquez"'
Search Results
2. Standardization and validation of a novel reverse transcriptase polymerase chain reaction method for detecting virulent strains of the infectious bursal disease virus
- Author
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Vladimir Longa-Bobadilla, Phillip Ormeño-Vásquez, Manuel Criollo-Orozco, Luis Tataje-Lavanda, Katherine Huamán-Gutierrez, Ángela Montalván, Mirko Zimic, Manolo Fernández-Sanchez, and Manolo Fernández-Díaz
- Subjects
alignment of sequences ,infectious bursal disease virus ,novel reverse transcriptase polymerase chain reaction ,virulent strains ,vp5 gene ,Animal culture ,SF1-1100 ,Veterinary medicine ,SF600-1100 - Abstract
Background and Aim: Gumboro disease is an economically crucial veterinary condition in chickens. It is caused by the infectious bursal disease virus (IBDV). This virus consists of two serotype groups, of which serotype I strain is pathogenic to chickens. For many years, the development of molecular techniques for either diagnostic purposes or surveillance of the appearance of new pathogenic strains has mainly focused on targeting the VP2 genomic region. However, due to the constant necessity for the discrimination between already prevalent vaccine strains and new pathogenic strains of this virus, it becomes imperative to have an immediate molecular method targeting a consensus sequence to achieve this task using field samples to reduce costs. Consequently, we focused on developing a novel reverse transcriptase polymerase chain reaction (RT-PCR) procedure solely for this purpose. Materials and Methods: Eight VP5 sequences were aligned, and the sequence with the majority of nucleotide coincidences was used to design a set of consensus primers. Then, a pathogenic strain of IBDV was propagated in embryonated chicken eggs, and the viral RNA was extracted. Finally, the conditions for this novel RT-PCR were evaluated using a commercial kit and the newly designed primers. Results: After determining the optimal RT-PCR conditions, the newly designed primers successfully amplified a 402-bp consensus sequence of the VP5 gene. In addition, these primers specifically amplified the VP5 sequence of the IBDV-positive samples, not the other samples previously confirmed to be positive for other common poultry pathogens. Conclusion: Our novel RT-PCR procedure has been demonstrated to be helpful in selectively amplifying the consensus sequence of the VP5 gene, indicating that this novel RT-PCR procedure constitutes an important and useful tool to execute initial discrimination of field-retrieved samples containing and not containing virulent strains of this virus before deciding to execute a blindly and more costly sequencing procedure of all the samples together.
- Published
- 2025
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- View/download PDF
3. Draft Genome Sequence of Heavy Metal-Resistant Aeromonas veronii CTe-01, Isolated from a Peruvian Wastewater Treatment Plant
- Author
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Lucy Espinoza-Salazar, Luis Tataje-Lavanda, Phillip Ormeño-Vásquez, Manolo Fernández-Díaz, Manolo Fernández-Sánchez, Mirko Zimic, Juan C. Tantaleán, Rosa Altamirano-Díaz, and Claudio C. Vásquez
- Subjects
Whole genome sequencing ,0303 health sciences ,biology ,Strain (chemistry) ,030306 microbiology ,Genome Sequences ,biology.organism_classification ,Microbiology ,03 medical and health sciences ,Bioremediation ,Immunology and Microbiology (miscellaneous) ,Genetics ,Sewage treatment ,Molecular Biology ,Bacteria ,030304 developmental biology ,Aeromonas veronii - Abstract
Here, we report a draft genome sequence of Aeromonas veronii strain CTe-01 (4.5 Mb), a hemolytic, heavy metal-resistant bacterium isolated from a wastewater treatment plant located at Cachiche, Ica, Peru. These characteristics could be used for bioremediation of contaminated environments.
- Published
- 2019
- Full Text
- View/download PDF
4. Desarrollo de una técnica de PCR para la detección de Fowl adenovirus 4 (FADV-4) en muestras de tejidos
- Author
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Vladimir Longa-Bobadilla, Phillip Ormeño-Vásquez, Víctor Chávez-Montenegro, Luis Alberto Tataje Lavanda, and Manolo Fernández-Díaz
- Subjects
General Veterinary - Abstract
Es importante contar con una técnica de PCR capaz de detectar adenovirus aviar 4 con una alta sensibilidad y precisión en cultivos celulares y muestras de origen aviar. En el presente trabajo se estandarizó una técnica de PCR. Primero se procedió a la preparación del Material de Referencia Interna (M.R.I.), usando el cultivo celular en células EB66, infectado FAdV-4; luego se purificó el virus, se extrajo el ADN y se cuantificó el amplicón. Finalmente se procedió a realizar la prueba de la gradiente de temperatura, prueba de concentración de cebadores, prueba de sensibilidad y prueba de especificidad utilizando el kit de PCR Q5® High-Fidelity 2X Master. De los cinco cebadores utilizados, los cebadores HHS*-3 y HHS*-4 pasaron todas las pruebas, siendo HHS*-3 el más sensible ya que detectó hasta 1 pg/µl.
- Published
- 2021
- Full Text
- View/download PDF
5. Near-Complete Genome Sequence of Infectious Bronchitis Virus Strain VFAR-047 (GI-16 Lineage), Isolated in Peru
- Author
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Luis Tataje-Lavanda, Katherine Huamán-Gutiérrez, Manolo Fernández-Díaz, Mirko Zimic-Peralta, Ray Izquierdo-Lara, and Phillip Ormeño-Vásquez
- Subjects
Genetics ,Whole genome sequencing ,0303 health sciences ,Lineage (genetic) ,040301 veterinary sciences ,Sequence analysis ,Strain (biology) ,Genome Sequences ,Infectious bronchitis virus ,social sciences ,04 agricultural and veterinary sciences ,Biology ,Genome ,0403 veterinary science ,03 medical and health sciences ,Immunology and Microbiology (miscellaneous) ,parasitic diseases ,population characteristics ,Molecular Biology ,human activities ,Recombination ,geographic locations ,030304 developmental biology - Abstract
Here, we report the near-complete genome sequence of the infectious bronchitis virus (IBV) strain VFAR-047, isolated in Peru in 2014. This strain was classified into GI lineage 16 (GI-16) based on both the genome and Spike 1 (S1) sequence analysis., Here, we report the near-complete genome sequence of the infectious bronchitis virus (IBV) strain VFAR-047, isolated in Peru in 2014. This strain was classified into GI lineage 16 (GI-16) based on both the genome and Spike 1 (S1) sequence analysis. Furthermore, four potential recombination events with other GI-16 and GI-11 strains were identified.
- Published
- 2019
6. Development of a molecular platform for the detection and quantification of Newcastle vaccine virus
- Author
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Vladimir Longa Bobadilla, Luis Tataje Lavanda, Phillip Ormeño Vásquez, Manuel Criollo Orozco, Katherine Calderón, Jorge Bendezú, Manolo Fernandez Díaz, Katherine Huamán Gutiérrez, Edison Huaccachi Gonzáles, and Angela Montalvan
- Subjects
Virus quantification ,RT-PCRc ,General Veterinary ,Hemagglutination ,biology ,RT-qPCR ,Embryonated ,embryonated SPF eggs ,huevos embrionados SPF ,medicine.disease ,biology.organism_classification ,Virology ,Newcastle disease ,Reverse transcriptase ,Virus ,law.invention ,NDV ,law ,medicine ,Polymerase chain reaction ,Ornithobacterium rhinotracheale - Abstract
El estudio tuvo por objetivo desarrollar una plataforma molecular para la cuantificación del virus de la enfermedad de Newcastle (NDV) a partir de un sistema de cultivo en huevos embrionados SPF. Primero se evaluaron cuatro pares de cebadores que amplifican diferentes regiones del genoma viral de NDV que codifican para la proteína de nucleocápside (NP), proteína matriz (M), proteína fusión (F) y la ARN polimerasa ARN dependiente (L), con la finalidad de seleccionar el más conservado a partir del cual se desarrolló una plataforma molecular basada en la transcripción reversa - reacción en cadena de polimerasa convencional (RT-PCRc) en dos pasos para la detección del NDV. Posteriormente, esta fue llevada a transcripción reversa - reacción en cadena de polimerasa en tiempo real (RT-qPCR) para la cuantificación de NDV producido a partir de un sistema de huevos embrionados. Mediante estas técnicas se determinó que los cebadores para el gen M fueron adecuados según los criterios de optimización para el desarrollo de ambos métodos. Mediante ensayos de sensibilidad se demostró que la RT-qPCR (116 copias genómicas/μl) era 10 veces más sensible que el RT-PCRc. Los cebadores fueron específicos pues no hubo amplificados en los controles negativos ni en otros patógenos aviares (virus de la laringotraqueitis infecciosa, metapneumovirus aviar, virus de la bronquitis infecciosa, Avibacterium paragallinarum, Gallibacterium anatis y Ornithobacterium rhinotracheale). Debido su sensibilidad y especificidad, se propone esta plataforma para la cuantificación de NDV vacunal cuando es producido a partir de un sistema de huevos embrionados, como una alternativa frente a métodos convencionales de titulación como hemaglutinación, ensayo en placa, TCDI50 y DIEP50., The objective of the study was to develop a molecular platform for the quantification of Newcastle disease virus (NDV) from a culture system in embryonated SPF eggs. First, four pairs of primers were evaluated that amplify different regions of the NDV viral genome that code for: nucleocapsid protein (NP), protein matrix (M), fusion protein (F) and RNA-dependent RNA polymerase (L) to select the most conserved one from which a molecular platform based on reverse transcription was developed - conventional polymerase chain reaction (RT-PCRc) in two steps for the detection of NDV. Subsequently, it was taken to reverse transcription - real-time polymerase chain reaction (RT-qPCR) for the quantification of NDV produced from a system of embryonated eggs. Through these techniques, it was determined that the primers for the M gene were adequate according to the optimization criteria for the development of both methods. Sensitivity tests showed that the RT-qPCR (116 genomic copies/μl) was 10 times more sensitive than the RT-PCRc. The primers proved to be specific since there were no amplifications in the negative controls or in other avian pathogens (infectious laryngotracheitis virus, avian metapneumovirus, infectious bronchitis virus, Avibacterium paragallinarum, Gallibacterium anatis and Ornithobacterium rhinotracheale). Due to its sensitivity and specificity, this platform is proposed for the quantification of NDV vaccine when it is produced from an embryonated egg system, as an alternative to conventional titration methods such as hemagglutination, plaque assay, TCDI50 and DIEP50.
- Published
- 2018
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