Comparative evaluation of conventional RT-PCR and real-time RT-PCR (RRT-PCR) for detection of avian metapneumovirus subtype A

Abstract

Avian metapneumovirus (AMPV) belongs to Metapneumovirus genus of Paramyxoviridae family. Virus isolation, serology, and detection of genomic RNA are used as diagnostic methods for AMPV. The aim of the present study was to compare the detection of six subgroup A AMPV isolates (AMPV/A) viral RNA by using different conventional and real time RT-PCR methods. Two new RT-PCR tests and two real time RT-PCR tests, both detecting fusion (F) gene and nucleocapsid (N) gene were compared with an established test for the attachment (G) gene. All the RT-PCR tested assays were able to detect the AMPV/A. The lower detection limits were observed using the N-, F-based RRT-PCR and F-based conventional to 10 1 TCID 50 mL -1 ). The present study suggests that the conventional F-based RT-PCR presented similar detection limit when compared to N-and F-based RRT-PCR and they can be successfully used for AMPV/A detection.

An affordable detection system based on RT-LAMP and DNA-nanoprobes for avian metapneumovirus

“…Currently, RT-PCR and qRT-PCR tests have become the established gold standard methods in reference laboratories for diagnosing active infections. Most of the RT-PCR epidemiological studies on aMPV are based on M, N, and G genes (Ferreira et al 2009 ; Kariithi et al 2022 ; Wang et al 2022 ). In this study, we targeted its F gene based on its high conservation degree in the reference strains of aMPV subtypes A and B.…”

Development of a Fast and Affordable Diagnostic System for RNA Viruses Using Loop-Mediated Isothermal Amplification and DNA-Nanoprobe Detection

“…Currently, RT-PCR and qRT-PCR tests have become the established gold standard methods in reference laboratories for diagnosing active infections. Indeed, primer sequences for the RT-PCR have been designed for specific detection of the F, M, N and G genes (Ferreira et al, 2009;Kariithi et al, 2022;Wang et al, 2022). For instance, since the gene G shows the highest variability between subtypes it is the region most widely used at aMPV genotyping.…”

Complete Genome Sequence of an Avian Metapneumovirus Subtype A Strain Isolated from Chicken (Gallus gallus) in Brazil

“…The tracheal tissue sample used in this study was obtained from a Brazilian commercial layer flock showing swollen head syndrome, previously detected by reverse transcription (RT)-PCR (4) and confirmed by real-time RT-PCR (5). Samples were collected according to international, national, and institutional guidelines for the care and use of animals.…”

“…Avian metapneumovirus (family Pneumoviridae , genus Metapneumovirus ) ( 1 ) is the viral agent responsible for rhinotracheitis of turkeys and is associated with swollen head syndrome in chickens ( 2 , 3 ). The tracheal tissue sample used in this study was obtained from a Brazilian commercial layer flock showing swollen head syndrome, previously detected by reverse transcription (RT)-PCR ( 4 ) and confirmed by real-time RT-PCR ( 5 ). Samples were collected according to international, national, and institutional guidelines for the care and use of animals.…”