tetano
Editor, Senior Moderator
J Virol Methods. 2017 Nov 9. pii: S0166-0934(17)30494-9. doi: 10.1016/j.jviromet.2017.11.004. [Epub ahead of print]
[h=1]RNAlater? is a viable storage option for avian influenza sampling in logistically challenging conditions.[/h] Wille M[SUP]1[/SUP], Yin H[SUP]2[/SUP], Lundkvist ?[SUP]3[/SUP], Xu J[SUP]4[/SUP], Muradrasoli S[SUP]5[/SUP], J?rhult JD[SUP]6[/SUP].
[h=3]Author information[/h]
[h=3]Abstract[/h] Surveillance of wild birds is critical in monitoring for highly pathogenic avian influenza A viruses (AIV). However, a successful surveillance regime requires proper treatment of samples in the field - rapid placement of samples in -80?C and subsequent maintenance of cold-chain. Given the logistical difficulties of this, many avian taxa and/or geographic locations are not sampled, or, when sampled may result in false negatives due to poor sample treatment in the field. Here, we assessed the utility of RNAlater[SUP]?[/SUP] as a stabilization agent for AIV sampling. We found no difference in real time PCR performance between virus transport media at optimal conditions and RNAlater[SUP]?[/SUP] at -80?C, -20?C, 4?C or room temperature up to two weeks, at either low or high virus load. Not only is RNAlater[SUP]?[/SUP] useful in comparison of spiked samples or those from duck experiments, it was employed successfully in a field study of backyard birds in China. We detected AIV in cloacal and oropharyngeal samples from chickens and a sample with a low Cq were successfully subtyped as H9, although sample storage conditions were suboptimal. Thus, despite limitations in downstream characterization such virus isolation and typing, RNAlater[SUP]?[/SUP] is a viable option for AIV screening under logistically challenging circumstances.
Copyright ? 2017. Published by Elsevier B.V.
[h=4]KEYWORDS:[/h] China; Influenza A virus; RNA preservative; RNAlater(?); Storage conditions; VTM
PMID: 29129490 DOI: 10.1016/j.jviromet.2017.11.004
[h=1]RNAlater? is a viable storage option for avian influenza sampling in logistically challenging conditions.[/h] Wille M[SUP]1[/SUP], Yin H[SUP]2[/SUP], Lundkvist ?[SUP]3[/SUP], Xu J[SUP]4[/SUP], Muradrasoli S[SUP]5[/SUP], J?rhult JD[SUP]6[/SUP].
[h=3]Author information[/h]
[h=3]Abstract[/h] Surveillance of wild birds is critical in monitoring for highly pathogenic avian influenza A viruses (AIV). However, a successful surveillance regime requires proper treatment of samples in the field - rapid placement of samples in -80?C and subsequent maintenance of cold-chain. Given the logistical difficulties of this, many avian taxa and/or geographic locations are not sampled, or, when sampled may result in false negatives due to poor sample treatment in the field. Here, we assessed the utility of RNAlater[SUP]?[/SUP] as a stabilization agent for AIV sampling. We found no difference in real time PCR performance between virus transport media at optimal conditions and RNAlater[SUP]?[/SUP] at -80?C, -20?C, 4?C or room temperature up to two weeks, at either low or high virus load. Not only is RNAlater[SUP]?[/SUP] useful in comparison of spiked samples or those from duck experiments, it was employed successfully in a field study of backyard birds in China. We detected AIV in cloacal and oropharyngeal samples from chickens and a sample with a low Cq were successfully subtyped as H9, although sample storage conditions were suboptimal. Thus, despite limitations in downstream characterization such virus isolation and typing, RNAlater[SUP]?[/SUP] is a viable option for AIV screening under logistically challenging circumstances.
Copyright ? 2017. Published by Elsevier B.V.
[h=4]KEYWORDS:[/h] China; Influenza A virus; RNA preservative; RNAlater(?); Storage conditions; VTM
PMID: 29129490 DOI: 10.1016/j.jviromet.2017.11.004