Development of loop mediated isothermal amplification (LAMP): A new tool for rapid diagnosis of cotton leaf curl viral disease.

Autor: Gawande SP; ICAR-Central Institute for Cotton Research (ICAR-CICR), Nagpur 440010, India. Electronic address: spgawande1@gmail.com., Raghavendra KP; ICAR-Central Institute for Cotton Research (ICAR-CICR), Nagpur 440010, India., Monga D; ICAR, Central Institute for Cotton Research Regional Station (ICAR-CICR RS), Sirsa 125055, India., Nagrale DT; ICAR-Central Institute for Cotton Research (ICAR-CICR), Nagpur 440010, India., Prabhulinga T; ICAR-Central Institute for Cotton Research (ICAR-CICR), Nagpur 440010, India., Hiremani N; ICAR-Central Institute for Cotton Research (ICAR-CICR), Nagpur 440010, India., Meshram M; ICAR-Central Institute for Cotton Research (ICAR-CICR), Nagpur 440010, India., Kranthi S; International Cotton Advisory Committee (ICAC), Washington, DC, USA., Gokte-Narkhedkar N; ICAR-Central Institute for Cotton Research (ICAR-CICR), Nagpur 440010, India., Waghmare VN; ICAR-Central Institute for Cotton Research (ICAR-CICR), Nagpur 440010, India.
Jazyk: angličtina
Zdroj: Journal of virological methods [J Virol Methods] 2022 Aug; Vol. 306, pp. 114541. Date of Electronic Publication: 2022 May 11.
DOI: 10.1016/j.jviromet.2022.114541
Abstrakt: Cotton leaf curl disease (CLCuD) ranks top among all endemic diseases transmitted by whitefly (Bemisia tabaci) affecting cotton (Gossypium hirsutum) causing severe economic losses to the cotton growers in the Indian subcontinent. For its effective management, robust tools for detection are a prerequisite and it is important to diagnose the virus titre in early stage of infection in plants as well as in the disease transmitting vector. Considering the limitations in current PCR-based techniques we have standardised rapid and sensitive Loop Mediated Isothermal Amplification (LAMP) protocol for the diagnosis of cotton leaf curl virus (CLCuV) in cotton leaves and in its transmitting vector whitefly. Perhaps, this is the first report of use of LAMP tool for rapid diagnosis of CLCuV in cotton and its transmitting vector the whitefly. Further, the colorimetric detection for diagnostic simplicity of amplified LAMP product by using different dyes lead to enhanced applicability of this technique in the field of disease diagnostics. The merit of present study is that the diagnostic failure of PCR and LAMP due to low virus titre in the infected leaf has been circumvented through the combination of rolling circle amplification (RCA) with LAMP. Thus RCA-LAMP can be an option for ultra-sensitive detection of samples with low virus titre. The potential applications of this advanced diagnostic tool in laboratory research on diagnosis of CLCuV, an important viral pathogen of cotton have been discussed.
(Copyright © 2022 Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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