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One-step SYBR green-based real-time RT-PCR assay for detection of foot-and-mouth disease virus circulating in India

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Title One-step SYBR green-based real-time RT-PCR assay for detection of foot-and-mouth disease virus circulating in India
Not Available
 
Creator Jitendra Kumar Biswal
Biswa Ranjan Jena
Syed Zeeshan Ali
Rajeev Ranjan
Jajati K. Mohapatra
Rabindra Prasad Singh
 
Subject FMDV · SYBR green · Real-time PCR · Diagnosis
 
Description Not Available
Rapid, sensitive, and reliable laboratory detection of foot-and-mouth disease virus (FMDV) infection is essential for containing and controlling virus infection in any geographical area. In this report a SYBR green-based 3Dpol-specific one-step real-time RT-PCR (rRT-PCR) assay was developed for the pan-serotype detection of FMDV in India. The detection limit of the SYBR green-based rRT-PCR was 10−2 TCID50/50 µl, which is 10 times more sensitive than the traditional agarose gel electrophoresis-based RT-multiplex PCR (RT-mPCR). The standard curve exhibited a linear range across 8-log10 units of viral RNA dilution. The reproducibility and specificity of this assay were reasonably high suggesting that the 3Dpol-specific SYBR green rRT-PCR could detect FMDV genome specifically and with little run-to-run variation. The new 3Dpol-specific SYBR green rRT-PCR assay was evaluated alongside the established RT-mPCR using the archived FMDV isolates and clinical field samples from suspected FMD outbreaks. A perfect concordance was observed between the new rRT-PCR and the traditional RT-mPCR on viral RNA in the archived FMDV cell culture isolates tested. Furthermore, 73% of FMDV-suspected clinical samples were detected positive through the 3Dpol-specific SYBR green rRT-PCR, while the detection rate through the traditional RT-mPCR was 57%. Therefore, the SYBR green-based 3Dpol-specific one-step rRT-PCR could be considered as a valuable assay with higher diagnostic sensitivity to complement the routine assays that are being used for FMD virus diagnosis in India.
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Date 2022-01-14T08:30:41Z
2022-01-14T08:30:41Z
2022-01-06
 
Type Research Paper
 
Identifier Biswal, J.K., Jena, B.R., Ali, S.Z. et al. One-step SYBR green-based real-time RT-PCR assay for detection of foot-and-mouth disease virus circulating in India. Virus Genes (2022). https://doi.org/10.1007/s11262-021-01884-3
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http://krishi.icar.gov.in/jspui/handle/123456789/68670
 
Language English
 
Relation Not Available;
 
Publisher Springer Nature