Development of Calorimetric PCR Assay as Cost Effective, Direct Post PCR Readout Platform


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Authors

  • Natesan Pazhanivel Department of Animal Biotechnology, Madras Veterinary College, Chennai-600007, Tamil Nadu, India
  • Divya Magdalina Department of Veterinary Pathology, Madras Veterinary College, Chennai-600007, Tamil Nadu, India
  • Manoharan Parthiban Department of Animal Biotechnology, Madras Veterinary College, Chennai-600007, Tamil Nadu, India
  • Gopal Dhinakar Raj Centre for Animal Health Studies, Tamil Nadu Veterinary and Animal Sciences University, Madhavaram Milk Colony, Chennai-600051, Tamil Nadu, India
  • John Kirubaharan Department of Veterinary Microbiology, Madras Veterinary College, Chennai-600007, Tamil Nadu, India
  • Gopal Sathish Department of Animal Biotechnology, Madras Veterinary College, Chennai-600007, Tamil Nadu, India
  • Karnan Divya Manjari Department of Animal Biotechnology, Madras Veterinary College, Chennai-600007, Tamil Nadu, India

Keywords:

COLORIMETRIC PCR-post PCR development – naked eye

Abstract

The sophisticated expensive molecular laboratories are commonly present for detection of molecular template in given sample. But cost effective point of care diagnostic methodology without much costlier instruments at field level is the need of hour for molecular diagnostics. Here in this study an attempt was made through calorimetric pcr as simple post pCR development strategy for reading out of results by naked eye. The calorimetric PCR assay was demonstrated by using Herpesvirus of Turkey (HVT) referral strain (one of the vaccine strain against Mareks disease virus) in this study.  HVT specific primers were designed by using primer 3.0 software targeting US3 gene-specific to  HVT strain. The forward primer of the US3 gene which is specific to HVT was integrated with HRPzyme sequence (566bp) and amplified during the PCR. The PCR product including HRPzyme produced the distinguished blue colour with chromogenic substrates which was visualized by the naked eye. From these observations, simple, rapid colorimetric reaction based post PCR analysis was performed without using agarose gel electrophoresis or real-time methods employing fluorescent dyes.

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References

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Submitted

2023-12-15

Published

2023-12-15

How to Cite

Natesan Pazhanivel, Divya Magdalina, Manoharan Parthiban, Gopal Dhinakar Raj, John Kirubaharan, Gopal Sathish, & Karnan Divya Manjari. (2023). Development of Calorimetric PCR Assay as Cost Effective, Direct Post PCR Readout Platform. The Indian Veterinary Journal, 100(12), 21-24. https://epubs.icar.org.in/index.php/IVJ/article/view/146478