Nucleotide sequences of the F, L and G protein genes of two non-A/non-B avian pneumoviruses (APV) reveal a novel APV subgroup

Abstract

Sequence analysis was performed of all or part of the genes encoding the fusion (F), polymerase (L) and attachment (G) proteins of two French non-A/non-B avian pneumovirus (APV) isolates (Fr/85/1 and Fr/85/2). The two isolates shared at least 99.7% nt and 99.0% aa sequence identity. Comparison with the F genes from subgroup A, subgroup B or Colorado APVs revealed nt and aa identities of 70.0-80. 5% and 77.6-97.2%, respectively, with the L gene sharing 76.1% nt and 85.3% aa identity with that of a subgroup A isolate. The Fr/85/1 and Fr/85/2 G genes comprised 1185 nt, encoding a protein of 389 aa. Common features with subgroup A and subgroup B G proteins included an amino-terminal membrane anchor, a high serine and threonine content, conservation of cysteine residues and a single extracellular region of highly conserved sequence proposed to be the functional domain involved in virus attachment to cellular receptors. However, the Fr/85/1 and Fr/85/2 G sequences shared at best 56.6% nt and 31.2% aa identity with subgroup A and B APVs, whereas these isolates share 38% aa identity. Phylogenetic analysis of the F, G and L genes of pneumoviruses suggested that isolates Fr/85/1 and Fr/85/2 belong to a previously unrecognized APV subgroup, tentatively named D. G-based oligonucleotide primers were defined for the specific molecular identification of subgroup D. These are the first G protein sequences of non-A/non-B APVs to be determined.

Detection and differentiation of avian pneumoviruses (metapneumoviruses)

“…This resulted in amplification of the F gene, which is located between the M and M2 genes (Figure 1). Bäyon-Auboyer et al (2000) used a similar strategy to amplify the G gene of subtype D virus. In that instance, they identified regions of high amino acid identity between the L proteins of subtype A APV and human RS virus, and made a negativesense and a positive-sense oligonucleotide accordingly.…”

Design, Validation, and Absolute Sensitivity of a Novel Test for the Molecular Detection of Avian Pneumovirus

“…2,3 A simple cutoff standard plastic centrifuge tube has also been recommended by others. 1 However, when cutting plastic, a 90Њ flat, smooth cut surface is very difficult to obtain, and because of this, such a method requires standing the tube over a ring of Vaseline to seal the imperfections left by the sawing of the tube. In addition, with the latter method, the cells are scattered over a very large area.…”

“…Through the years, several methods that do not require a centrifuge have been proposed. [1][2][3][4][5] One of these uses a special device, consisting of several precisely assembled, machined, plastic and metal parts. 2,3 A simple cutoff standard plastic centrifuge tube has also been recommended by others.…”

“…This study considers the detection by PCR of APV, a virus that has frequently proved a difficult agent to detect by conventional isolation Dipartimento di Sanità Pubblica Veterinaria e Patologia Animale, Facoltà di Medicina Veterinaria, Università di Bologna, Via Tolara di Sopra, 50, 40064 Ozzano Emilia (BO), Italy (Cecchinato, Catelli), and the Department of Veterinary Pathology, Jordan Building, Leahurst, Neston, Cheshire, CH64 7TE, UK (Savage, Jones, Naylor). 1 Corresponding Author: Elena Catelli, Dipartimento di Sanità Pubblica Veterinaria e Patologia Animale, Facoltà di Medicina Veterinaria, Università di Bologna, Via Tolara di Sopra, 50, 40064 Ozzano Emilia (BO), Italy.…”

“…8 The last subtype, referred to as D, includes 2 old French isolates. 1 Polymerase chain reaction (PCR) has been proved an invaluable tool in the detection of infectious agents since its inception in 1983. It has become possible to rapidly detect viral nucleic acid in a minimal time period, and amplified products can frequently be sequenced to provide more information about the etiological agent.…”