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Application of LAMP method for detection and quantification of Newcastle disease virus

Newcastle disease is one of highly contagious viral infectious diseases of domestic poultry and wild birds with a high morbidity and mortality and expensive control, suppression and eradication measures. To apply appropriate measures on time, due to its devastating effects on poultry industry, it needs rapid, simple and reliable diagnostic method. The aim of this work is to explore the possibility of using loop-mediated isothermal amplification (LAMP) in detection and absolute quantification of Newcastle disease virus in clinical samples. Reverse transcription of viral RNA, amplification and detection of target sequences was performed in one step at a constant temperature during the reaction in a RealTime PCR device. Real time monitoring of LAMP has shown positive reaction in 6, 5 to 14 minutes and enabled the approximate absolute quantification of initial viral nucleic acid concentration of a sample based on the standard curve. RT-LAMP has proved itself to be a simple, rapid, specific and sensitive method of nucleic acid amplification as well as quantification.

Newcastle disease virus; qRT-LAMP; qRT-PCR; absolute quantification

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