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Original Research
10.
Nested RT-PCR for ante mortem diagnosis of rabies
from body secretion/excretion of animals suspected
for rabies -
M Dandale, C K Singh, V Ramneek, D Deka, B S Sandhu,
K Bansal, N K Sood
Vet World. 2012; 5(11): 690-693
doi:
10.5455/vetworld.2012.
690-693
Abstract
Aim:
The present study deals with molecular technique
Nested RT-PCR for detection of rabies viral RNA from
biological fluid samples (Saliva, Milk and Urine)
collected from animal suspected for rabies and to
compare the sensitivity of Nested RT-PCR applied for
ante mortem diagnosis of rabies with conventional
technique (immunofluorescence) applied on neural
tissue.
Materials and Methods: Nested RT-PCR
was applied on 62 biological fluid specimens
collected from rabies suspected animals. First round
amplification with nested set of primers (RabN1 and
RabN5) yielded 1477 bp product while amplification
with second round primers (RabNfor and RabNrev)
yielded 762 bp product. Sensitivity of the technique
was compared in accordance with WHO recommended gold
standard test viz. Immunofluorescence (FAT) applied
on brain samples.
Results: By Nested RT-PCR,
viral RNA could be detected in 9/24 (37.50%) saliva
samples, 2/17 (11.76%) milk samples and 6/21
(28.57%) urine samples. Confirmatory diagnosis by
Immunofluorescence performed on brain sample
revealed 18 true positive cases. Overall,
Sensitivity of Nested RT-PCR technique employed on
fluid samples was 69.23% when compared with
immunofluorescence performed on brain samples.
Conclusions: Early reliable ante mortem
diagnosis of rabies can be obtained from biological
fluid samples of animals suspected to be rabid when
tested with Nested RT-PCR technique.
Keywords:
milk,nested RT-PCR, rabies, saliva, urine
