Comparative detection of foot-and-mouth disease virus by reverse transcription loop-mediated isothermal amplification assay and real time polymerase chain reaction in Uganda
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Date
2016
Journal Title
Journal ISSN
Volume Title
Publisher
Int J Biotechnol Food Sci
Abstract
Foot-and-mouth disease (FMD) is a viral disease. FMD diagnosis in the field is based on clinical signs that
are shared by other vesicular diseases, hence to confirm FMD a laboratory is needed. Laboratory diagnostic techniques
including serology may fail to distinguish between vaccinated and new infection, virus isolation may take up to 4 days to
yield results, while molecular techniques including PCR, which are accurate, sensitive, specific and rapid, are costly and
require special training of the laboratory staff. These challenges limit laboratory diagnosis yet in Uganda FMD outbreaks
are common since the disease is endemic. This work reports the comparative detection of Foot-and-mouth disease virus
(FMDV) by reverse transcription-loop mediated isothermal amplification (RT-LAMP) and Real-Time polymerase chain
reaction (rRT-PCR) in Uganda based on the 3D polymerase (3Dpol) gene. The rRT-PCR assay is considered as the gold
standard. A total of 89 cattle samples that included epithelial tissues (16.9%) and oral swabs (83.1%) were collected
from outbreak cases in Eastern Districts of Mbale and Budaka. These were applied to molecular assays of rRT-PCR and
RT- LAMP using primers and probes targeting the 3Dpol gene. The diagnostic sensitivity and specificity of RT-LAMP as a
screening test and rRT-PCR as the reference test was 94.44% (95% CI = 94.11 to 94.78%) and 98.59% (95% CI =
98.50 to 98.68%), respectively. The kappa value for diagnostic agreement between rRT-PCR and RT-LAMP was 93.0%
(95% CI = 83.50 to 100%), showing a perfect agreement. In conclusion, the RT-LAMP assay had a very high sensitivity
and specificity when compared to the reference test of rRT-PCR. It was also very rapid since it gave results in 45 to 60
min. Due to its simplicity, sensitivity and specificity, LAMP assay has the potential for use in routine surveillance of FMD
in Uganda.
Description
Keywords
Foot-and-mouth disease virus, real-time polymerase chain reaction, reverse transcription LAMP
Citation
Mukasa, H. K., Mwiine, F. N., Atuhaire, D. K., Ochwo, S., & Nanteza, A. (2016). Comparative detection of foot-and-mouth disease virus by reverse transcription loop-mediated isothermal amplification assay and real time polymerase chain reaction in Uganda. Int J Biotechnol Food Sci, 4, 22-33.