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

Author(s)

Ferreira, Helena Lage ^[1] ; Spilki, Fernando Rosado ^[2] ; Aparecida Bianchi dos Santos, Marcia Merces ^[3] ; de Almeida, Renata Servan ^[4] ; Arns, Clarice Weis ^[3]

Affiliation

^[1] Vet & Agrochem Res Inst, B-1180 Brussels - Belgium

^[2] Ctr Univ Feevale, Inst Ciencias Saude, Novo Hamburgo, RS - Brazil

^[3] Univ Estadual Campinas, Dept Microbiol & Imunol, Inst Biol, Lab Virol Anim, BR-06109 Campinas, SP - Brazil

^[4] CIRAD, Dept Syst Biol, Unite Controle Malad Anim Exot & Emergentes, F-34398 Montpellier - France

• Document type: Journal article
• Source: Ciência Rural; v. 39, n. 5, p. 1445-1451, AUG 2009.
• Web of Science Citations: 5

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 RT-PCR (10(0.3) 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. (AU)

• FAPESP's process: 03/14012-9 - The effect of RNAi application in avian pneumovirus infection
• Grantee: Helena Lage Ferreira
• Support type: Scholarships in Brazil - Doctorate (Direct)