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.

ABSTRACT

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.

To Access the original article, check the following link: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5843735/