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One-step multiplex RT-PCR assay for the detection of peste des petits ruminants virus in clinical samples

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Title One-step multiplex RT-PCR assay for the detection of peste des petits ruminants virus in clinical samples
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Creator V Balamurugan, A Sen, P Saravanan, R P Singh, R K Singh, T J Rasool, S K Bandyopadhyay
 
Subject one-step multiplex RT-PCR, peste des petits ruminants virus, differentiation RPV, clinical samples
 
Description Not Available
A single-tube one-step multiplex RT-PCR was standardized to amplify both 337 bp and 191 bp fragments of
N and M genes of peste des petits ruminants virus (PPRV), respectively, and only a 337 bp fragment of N gene
of Rinderpest virus (RPV). The RT-PCR using purified viral RNA was easily adopted for direct detection of
PPRV in clinical field samples and its differentiation from RPV. The amplified N and M gene products were
confirmed to be PPRV- and RPV-specific by their size in 1.5% agarose gel and restriction analysis. In the
assay, the Qiagen one-step RT-PCR kit containing the Ominiscript and Sensiscript reverse transcriptases and
Hot star Taq DNA polymerase was utilized. The sensitivity of the assay was found to be 100 fg of PPRV RNA.
Compared with a two-step assay, the one-step assay is easier and time-saving as it requires just a single buffer
for both reactions, reverse transcription (RT) and PCR. In experimentally infected goats, PPRV was detectable
by the one-step RT-PCR in nasal and ocular swabs 7–17 days post infection (p.i.). and in oral swabs 7–15 days
p.i. Out of 32 clinical field samples tested, 18 were positive by sandwich ELISA (S-ELISA), while 22 were
positive by the one-step RT-PCR.
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Date 2023-05-30T08:47:47Z
2023-05-30T08:47:47Z
2006-07
 
Type Research Paper
 
Identifier Balamurugan V, Sen A, Saravanan P, Singh RP, Singh RK, Rasool TJ, Bandyopadhyay SK. One-step multiplex RT-PCR assay for the detection of peste des petits ruminants virus in clinical samples. Vet Res Commun. 2006 Aug;30(6):655-66. doi: 10.1007/s11259-006-3331-3. PMID: 16838207.
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http://krishi.icar.gov.in/jspui/handle/123456789/77898
 
Language English
 
Relation Not Available;
 
Publisher SpringerLink