Abstract
The 2009 pandemic A (H1N1) influenza virus was first identified in Mexico in April 2009 and spread world wide over a short period of time. Well validated diagnostic methods that are rapid and sensitive for detection and tracking of this virus are urgently needed. In this study, time course kinetic characterizations of the abundances of all ten genes of the 2009 pandemic A (H1N1) influenza virus in standard virions and infected Madin-Darby Canine Kidney (MDCK) cells were monitored. Results showed that the amounts of each gene in infected cells were significantly higher than those in virions, so that cell lysates were more recommended to be the nucleotide materials detection object than virions. Meanwhile, all genes were present in virions in approximately equimolar amounts, whereas the copy numbers of each gene in cell lysates were distinguishing. The abundances of M1 and NP genes were highest and may be the optimized choice for nucleotide detection in infected cells. Furthermore, the most sxensitive time point for viral nucleotide detection in cells was 48 to 56 h post infection. In infected MDCK cells, the total RNAs amounts of NP and NS1 genes began to rise at 3 h post infection, whereas other eight genes escalated from 8 h post infection just as the situation of all genes in standard virions. All these data may be useful for more sensitive diagnosis and surveillance of the novel A (H1N1) virus, and might further limit the transmission of this pandemic disease in the future. Key words: The 2009 pandemic (H1N1) influenza virus, real-time PCR, virions, Madin-Darby Canine Kidney (MDCK) cells, abundance, sensitivity.
Highlights
On June 11, 2009, the World Health Organization raised the global pandemic alert level to phase 6, the pandemic phase, in response to the emergence and global spread of a novel influenza A (H1N1) virus, which emerged in Mexico in early 2009 (Garten et al 2009)
The WHO defines a probable clinical case as one that is confirmed by (1) specific real-time Polymerase chain reaction (PCR) based detection methods, (2) isolation of the pandemic A (H1N1) influenza virus, or (3) detection of 4-fold rise of neutralization antibodies to this virus (WHO, 2009)
BSL-2 laboratories with biosafety level 3 (BSL-3) practices are recommended for virus isolation and serology, whereas PCR detection requires only a BSL-2 environment
Summary
On June 11, 2009, the World Health Organization raised the global pandemic alert level to phase 6, the pandemic phase, in response to the emergence and global spread of a novel influenza A (H1N1) virus, which emerged in Mexico in early 2009 (Garten et al 2009). The transmissibility of this virus was estimated to be higher than that of seasonal influenza viruses (Fraser et al 2009). Most research and commercial detection kits for PCR diagnosis of the pandemic A (H1N1) influenza virus are targeted on Syntax Error: Unknown character collection 'Adobe-WinCharSetFFFF'
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
