Tataje-Lavanda L.1, Huamán-Gutierrez K.1, Longa-Bobadilla V.1, López G.2, Nolasco O.2, Bendezú J.1, Fernández-Díaz M.1.
1FARVET, Research and Development Laboratories, Ica, Peru. 2Universidad Peruana Cayetano Heredia, Instituto de Medicina Tropical “Alexander von Humboldt”, Lima, Peru.
In this study, we developed, standardized and validated a methodology following good laboratory practices (GLP) and the guidelines of the Food and Drug Administration (FDA) to measure the titer of vaccines produced in allantoic fluid (AF). A real-time PCR (qPCR) methodology was designed for the detection and quantification of the glycoprotein B gene (gB) of Avian infectious laryngotracheitis virus (ILTV) using SYBR Green I. The validation criteria showed that the qPCR assay has a limit of detection (LoD) of 1.017×105 genome copies/µL of AF which guarantees high precision, and a limit of quantification (LoQ) of 3.39×105 genome copies/µL of AF. These parameters demonstrated the safety and accuracy of the correct detection and quantification of the ILTV viral load in vaccines produced in AF, for the quality assurance of vaccines for birds and for diagnostic, virus load on positive samples.
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