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Open Access
Copyright: The authors. This article is an open access
article licensed under the terms of the Creative Commons Attribution License
(http://creativecommons.org/licenses/by/2.0) which permits unrestricted use,
distribution and reproduction in any medium, provided the work is properly
cited.
Original Research
3.
Use of reverse transcriptase loop-mediated
isothermal amplification assay for field detection
of newcastle disease virus using less invasive
samples -
H Kirunda, O M M Thekisoe, P D Kasaija, S D Kerfua,
G W Nasinyama, J Opuda-Asibo and N Inoue
Vet World. 2012; 5(4): 206-212
doi:
10.5455/vetworld.201 2.206-212
Abstract
A novel
nucleic acid amplification method, loop-mediated
isothermal amplification, was developed and
recently demonstrated detection of Newcastle
disease virus (NDV) in tissue samples. But
slaughter of poultry for test samples is often
faced with resentment by low-income farmers. This
study was undertaken to determine the test
properties of reverse transcriptase loop-mediated
isothermal amplification (RT-LAMP) in detection of
NDV in clinical cases using cloacal and
oropharyngeal swabs. Samples included 46 tracheal
tissues, 94 cloacal and 107 oro-pharyngeal swabs
from on-station and 30 spleens, 74 cloacal and 74
oro-pharyngeal swabs from the field. Analysis was
done using specific RT-LAMP targeting the fusion
(F) protein. While the method detected NDV from
swab samples, no RNA of other poultry disease
viruses was amplified, indicating analytical
specificity of 100%. RT-LAMP took ≤36 minutes in
83% (n=329) of positive reactions with all samples
amplified in <60 minutes. Results were easily
observed with a naked eye. Cloacal and oro-pharyngeal
swabs could be a convenient and cheaper
alternative in diagnosis of NDV infection by RT-LAMP
in resource poor countries.
Keywords:
Newcastle disease virus; RT-LAMP; sensitivity;
specificity; swab sample
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