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- Title
Comparative evaluation of nucleic acid-based assays for detection of Japanese encephalitis virus in swine blood samples.
- Authors
Dhanze, H.; Bhilegaonkar, K.; Ravi Kumar, G.; Thomas, P.; Chethan Kumar, H.; Suman Kumar, M.; Rawat, S.; Kerketta, P.; Rawool, D.; Kumar, A.
- Abstract
Japanese encephalitis is an emerging mosquito-borne flaviviral zoonotic disease. The present study was undertaken with the objective of developing rapid and sensitive nucleic-acid-based assays for detection of Japanese encephalitis virus (JEV) in swine blood samples. Three nucleic-acid-based assays, viz., reverse transcription polymerase chain reaction (RT-PCR), reverse transcription loop-mediated isothermal amplification (RT-LAMP), and real-time RT-PCR, were developed and compared in terms of their diagnostic efficacy. All three assays were found to be 100 per cent specific. The minimum detection limit of RT-LAMP and real-time RT-PCR was 12 copies/µl, while RT-PCR could detect 1.2 × 10 copies/µl. On comparison, RT-LAMP and real-time RT-PCR were 4-log more sensitive than RT-PCR. The applicability of the assays was evaluated by screening 135 field swine blood samples, of which 24 (17.77 %) were positive by RT-LAMP and real-time RT-PCR and only six (4.44 %) were positive by RT-PCR. The viral load in swine blood samples ranged between 2 × 10 and 4.8 × 10 copies per ml of blood by real-time RT-PCR. The comparative diagnostic sensitivity and specificity of RT-LAMP vis-à-vis real-time RT-PCR was found to be 100 %, while the sensitivity and specificity of RT-PCR vis-à-vis real-time RT-PCR was found to be 25 % and 100 %, respectively. Thus, the use of RT-PCR may cause the incidence of JEV in the swine population to be underestimated, while the real-time RT-PCR reported here is the test of choice for reference laboratories, and the newly developed one-step RT-LAMP assay will be suitable for field-level testing.
- Subjects
COMPARATIVE studies; JAPANESE encephalitis viruses; ZOONOSES; NUCLEIC acids; BLOOD sampling; MOSQUITO vectors; LABORATORY swine
- Publication
Archives of Virology, 2015, Vol 160, Issue 5, p1259
- ISSN
0304-8608
- Publication type
Article
- DOI
10.1007/s00705-015-2385-3