In vitro isolation and molecular identification of reptarenavirus in Malaysia

Boid inclusion body disease (BIBD) is a viral disease of boids caused by reptarenavirus. In this study, tissue from naturally infected boid snakes were homogenized and propagated in African Monkey kidney (Vero) and rat embryonic fibroblast (REF) cells. Virus replication was determined by the presenc...

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Main Authors: Abba, Yusuf, Abu Hassim, Hasliza, Hamzah, Hazilawati, Ibrahim, Omar Emad, Ilyasu, Yusuf, Bande, Faruku, Mohd Lila, Mohd Azmi, Mohamed Mustapha, Noordin
Format: Article
Language:English
Published: Springer 2016
Subjects:
Online Access:http://psasir.upm.edu.my/id/eprint/54538/
http://psasir.upm.edu.my/id/eprint/54538/1/In%20vitro%20isolation%20and%20molecular%20identification%20of%20reptarenavirus%20in%20Malaysia.pdf
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author Abba, Yusuf
Abu Hassim, Hasliza
Hamzah, Hazilawati
Ibrahim, Omar Emad
Ilyasu, Yusuf
Bande, Faruku
Mohd Lila, Mohd Azmi
Mohamed Mustapha, Noordin
author_facet Abba, Yusuf
Abu Hassim, Hasliza
Hamzah, Hazilawati
Ibrahim, Omar Emad
Ilyasu, Yusuf
Bande, Faruku
Mohd Lila, Mohd Azmi
Mohamed Mustapha, Noordin
author_sort Abba, Yusuf
building UPM Institutional Repository
collection Online Access
description Boid inclusion body disease (BIBD) is a viral disease of boids caused by reptarenavirus. In this study, tissue from naturally infected boid snakes were homogenized and propagated in African Monkey kidney (Vero) and rat embryonic fibroblast (REF) cells. Virus replication was determined by the presence of cytopathic effect, while viral morphology was observed using transmission electron microscopy. Viral RNA was amplified using RT-PCR with primers specific for the L-segment of reptarenavirus; similarly, quantification of viral replication was done using qPCR at 24–144 h postinfection. Viral cytopathology was characterized by cell rounding and detachment in both Vero and REF cells. The viral morphology showed round-to-pleomorphic particles ranging from 105 to 150 nm which had sand-like granules. Sanger sequencing identified four closely associated reptarenavirus species from 15 (37.5 %) of the total samples tested, and these were named as follows: reptarenavirus UPM-MY 01, 02, 03, and 04. These isolates were phylogenetically closely related to the University Helsinki virus (UHV), Boa Arenavirus NL (ROUTV; BAV), and unidentified reptarenavirus L20 (URAV-L20). Comparison of deduced amino acid sequences further confirmed identities to L-protein of UHV, L-polymerase of BAV and RNA-dependent RNA polymerase of URAV-L20. Viral replication in Vero cells increased steadily from 24 to 72 h and peaked at 144 h. This is the first study in South East Asia to isolate and characterize reptarenavirus in boid snakes with BIBD.
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spelling upm-545382018-03-27T06:15:15Z http://psasir.upm.edu.my/id/eprint/54538/ In vitro isolation and molecular identification of reptarenavirus in Malaysia Abba, Yusuf Abu Hassim, Hasliza Hamzah, Hazilawati Ibrahim, Omar Emad Ilyasu, Yusuf Bande, Faruku Mohd Lila, Mohd Azmi Mohamed Mustapha, Noordin Boid inclusion body disease (BIBD) is a viral disease of boids caused by reptarenavirus. In this study, tissue from naturally infected boid snakes were homogenized and propagated in African Monkey kidney (Vero) and rat embryonic fibroblast (REF) cells. Virus replication was determined by the presence of cytopathic effect, while viral morphology was observed using transmission electron microscopy. Viral RNA was amplified using RT-PCR with primers specific for the L-segment of reptarenavirus; similarly, quantification of viral replication was done using qPCR at 24–144 h postinfection. Viral cytopathology was characterized by cell rounding and detachment in both Vero and REF cells. The viral morphology showed round-to-pleomorphic particles ranging from 105 to 150 nm which had sand-like granules. Sanger sequencing identified four closely associated reptarenavirus species from 15 (37.5 %) of the total samples tested, and these were named as follows: reptarenavirus UPM-MY 01, 02, 03, and 04. These isolates were phylogenetically closely related to the University Helsinki virus (UHV), Boa Arenavirus NL (ROUTV; BAV), and unidentified reptarenavirus L20 (URAV-L20). Comparison of deduced amino acid sequences further confirmed identities to L-protein of UHV, L-polymerase of BAV and RNA-dependent RNA polymerase of URAV-L20. Viral replication in Vero cells increased steadily from 24 to 72 h and peaked at 144 h. This is the first study in South East Asia to isolate and characterize reptarenavirus in boid snakes with BIBD. Springer 2016-10 Article PeerReviewed text en http://psasir.upm.edu.my/id/eprint/54538/1/In%20vitro%20isolation%20and%20molecular%20identification%20of%20reptarenavirus%20in%20Malaysia.pdf Abba, Yusuf and Abu Hassim, Hasliza and Hamzah, Hazilawati and Ibrahim, Omar Emad and Ilyasu, Yusuf and Bande, Faruku and Mohd Lila, Mohd Azmi and Mohamed Mustapha, Noordin (2016) In vitro isolation and molecular identification of reptarenavirus in Malaysia. Virus Genes, 52 (5). pp. 640-650. ISSN 0920-8569; ESSN: 1572-994X https://link.springer.com/article/10.1007%2Fs11262-016-1345-7 Boid inclusion body disease; Reptarenavirus; Cell culture; Viral isolation; Sanger sequencing; Electron microscopy 10.1007/s11262-016-1345-7
spellingShingle Boid inclusion body disease; Reptarenavirus; Cell culture; Viral isolation; Sanger sequencing; Electron microscopy
Abba, Yusuf
Abu Hassim, Hasliza
Hamzah, Hazilawati
Ibrahim, Omar Emad
Ilyasu, Yusuf
Bande, Faruku
Mohd Lila, Mohd Azmi
Mohamed Mustapha, Noordin
In vitro isolation and molecular identification of reptarenavirus in Malaysia
title In vitro isolation and molecular identification of reptarenavirus in Malaysia
title_full In vitro isolation and molecular identification of reptarenavirus in Malaysia
title_fullStr In vitro isolation and molecular identification of reptarenavirus in Malaysia
title_full_unstemmed In vitro isolation and molecular identification of reptarenavirus in Malaysia
title_short In vitro isolation and molecular identification of reptarenavirus in Malaysia
title_sort in vitro isolation and molecular identification of reptarenavirus in malaysia
topic Boid inclusion body disease; Reptarenavirus; Cell culture; Viral isolation; Sanger sequencing; Electron microscopy
url http://psasir.upm.edu.my/id/eprint/54538/
http://psasir.upm.edu.my/id/eprint/54538/
http://psasir.upm.edu.my/id/eprint/54538/
http://psasir.upm.edu.my/id/eprint/54538/1/In%20vitro%20isolation%20and%20molecular%20identification%20of%20reptarenavirus%20in%20Malaysia.pdf