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Virology Journal

, 3:47

First Online: 19 June 2006Received: 16 January 2006Accepted: 19 June 2006

Abstract

BackgroundNipah virus is a zoonotic virus isolated from an outbreak in Malaysia in 1998. The virus causes infections in humans, pigs, and several other domestic animals. It has also been isolated from fruit bats. The pathogenesis of Nipah virus infection is still not well described. In the present study, Nipah virus replication kinetics were estimated from infection of African green monkey kidney cells Vero using the one-step SYBR Green I-based quantitative real-time reverse transcriptase-polymerase chain reaction qRT-PCR assay.

ResultsThe qRT-PCR had a dynamic range of at least seven orders of magnitude and can detect Nipah virus from as low as one PFU-μL. Following initiation of infection, it was estimated that Nipah virus RNA doubles at every ~40 minutes and attained peak intracellular virus RNA level of ~8.4 log PFU-μL at about 32 hours post-infection PI. Significant extracellular Nipah virus RNA release occurred only after 8 hours PI and the level peaked at ~7.9 log PFU-μL at 64 hours PI. The estimated rate of Nipah virus RNA released into the cell culture medium was ~0.07 log PFU-μL per hour and less than 10% of the released Nipah virus RNA was infectious.

ConclusionThe SYBR Green I-based qRT-PCR assay enabled quantitative assessment of Nipah virus RNA synthesis in Vero cells. A low rate of Nipah virus extracellular RNA release and low infectious virus yield together with extensive syncytial formation during the infection support a cell-to-cell spread mechanism for Nipah virus infection.

Electronic supplementary materialThe online version of this article doi:10.1186-1743-422X-3-47 contains supplementary material, which is available to authorized users.

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Autor: Li-Yen Chang - AR Mohd Ali - Sharifah Syed Hassan - Sazaly AbuBakar

Fuente: https://link.springer.com/







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