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A rapid and sensitive one step-SYBR green based semi quantitative real time RT-PCR for the detection of peste des petits ruminants virus in the clinical samples

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Virologica Sinica : Volume 26

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Virologica Sinica : Volume 22

A rapid and sensitive one step-SYBR green based semi quantitative real time RT-PCR for the detection of peste des petits ruminants virus in the clinical samples

Content Provider	Springer Nature Link
Author	Balamurugan, Vinayagamurthy Sen, Arnab Venkatesan, Gnanavel Yadav, Vinita Bhat, Vandna Bhanuprakash, Veerakyathappa Singh, Raj Kumar
Copyright Year	2012
Abstract	A sensitive and rapid single step real time (rt) RT-PCR was standardized using one-step Brilliant SYBR Green kit® for detection and semi-quantitation of peste des petitis ruminants virus (PPRV) using the virus RNA and matrix (M) protein gene-specific primers and compared with established conventional RT-PCR and TaqMan RT-PCR. The assay amplifies a 124 bp fragment of the PPRV M gene with T$_{m}$ of 78.28 to 78.50. The assay was linear within a range of 50 ng to 0.5 fg total virus RNA with a detection limit (sensitivity) of 0.5 fg. Based on the serial dilution of the live-attenuated PPR vaccine virus, the detection limit was ∼0.0001 cell culture infectious dose 50% units (TCID$_{50}$). Additionally, swab materials spiked with known titre of vaccine virus were equally well detected in the assay. The standardized rt RT-PCR was easily employed for the detection of PPRV nucleic acid directly in the field and experimental clinical samples. The assay detected the PPRV nucleic acid as early as 3 day post infection (dpi) and up to 20 dpi in swab materials from the experimental samples. The assay was rapid and more sensitive than TaqMan and conventional RT-PCR in the detection of PPRV nucleic acid from the PPR suspected clinical samples of sheep and goats. Therefore, the established, simplified SYBR green rt RT-PCR is an alternative test to the already existing various diagnostic assays and could be useful for rapid clinical diagnosis with advantage in reducing risk of contamination.
Starting Page	1
Ending Page	9
Page Count	9
File Format	PDF
ISSN	16740769
Journal	Virologica Sinica
Volume Number	27
Issue Number	1
e-ISSN	1995820X
Language	English
Publisher	SP Wuhan Institute of Virology, CAS
Publisher Date	2012-01-22
Publisher Place	Heidelberg
Access Restriction	Subscribed
Subject Keyword	PPR M gene SYBR green RT-PCR Early diagnosis Clinical samples Oncology Medical Microbiology Microbial Genetics and Genomics Microbiology Biochemistry Virology
Content Type	Text
Resource Type	Article
Subject	Infectious Diseases Virology Immunology Molecular Medicine

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A rapid and sensitive one step-SYBR green based semi quantitative real time RT-PCR for the detection of peste des petits ruminants virus in the clinical samples

Cytokines expression profile and kinetics of Peste des petits ruminants virus antigen and antibody in infected and vaccinated goats

One-step Multiplex RT-PCR Assay for the Detection of Peste des petits ruminants Virus in Clinical Samples

Development and Evaluation of a Nested PCR for Improved Diagnosis and Genetic Analysis of Peste des Petits Ruminants Virus (PPRV) for Future Use in Nascent PPR Eradication Programme

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PREVALENCE OF PESTE DES PETITS RUMINANTS (PPR) VIRUS IN SMALL RUMINANTS OF GUJARAT AND ITS CHARACTERIZATION BY RT - PCR / RFLP AND SSCP PROFILES

Confirmed diagnosis by RT-PCR and phylogenetic analysis of peste des petits ruminants viruses in Tibet, China

One-step Multiplex RT-PCR Assay for the Detection of Peste des petits ruminants Virus in Clinical Samples

ASSESSMENT OF NOVEL N-GENE TARGET FOR DETECTION OF PESTE DES PETITS RUMINANTS (PPR) VIRUS BY RT-PCR AND CHARACTERIZATION BY RFLP PROFILE

A rapid and sensitive one step-SYBR green based semi quantitative real time RT-PCR for the detection of peste des petits ruminants virus in the clinical samples

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