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Development of probe-based real-time loop-mediated isothermal amplification for detection of Brucella

I A Bhat¹, M Mashooq¹, D Kumar²

¹Division of Bacteriology & Mycology, Indian Veterinary Research Institute, Izatnagar, UP, India.

Journal of Applied Microbiology

|June 1, 2018

Abstract

Aim:

To develop a rapid, sensitive and specific diagnostic assay for the detection of Brucella.

Methods And Results:

The probe-based RT-LAMP was carried out by using a set of four or six primers and different LAMP chemicals to compare its results with real-time PCR. Detection of gene amplification is done within 40 min and can be seen by amplification curve, turbidity and addition of DNA-binding dye at the end of the reaction results in colour difference under normal day light and in UV. The sensitivity of probe-based real-time LAMP assay was found 10-fold higher than Taqman-based qPCR. The specificity of the developed assay was validated by the absence of any cross-reaction with other pathogenic bacteria.

Conclusion:

The developed probe-based RT-LAMP assay is extremely rapid, cost effective, highly specific and sensitive, and has potential usefulness for rapid Brucella surveillance.

Significance And Impact Of The Study:

The developed probe-based RT-LAMP is a powerful gene amplification technique which is a specific, fast diagnostic tool for early detection and identification of Brucella.

Keywords:

Brucella RT-LAMP Taqman amplification probe

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