A Field-Friendly Loop-Mediated Isothermal Amplification (FF-LAMP) method for rapid detection of Nosema bombycis in silkworm, Bombyx mori

Pebrine is a destructive disease that exhibits horizontal and vertical transmission and therefore it is the only mandatory quarantine item in sericulture. Here, a field-friendly loop-mediated isothermal amplification (FF-LAMP) method has been developed and validated for the rapid detection of Nosem...

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Autores principales:	V Sivaprasad, L Satish, G Mallikarjuna, N Chandrakanth, A V Mary Josepha, S M Moorthy
Formato:	article
Lenguaje:	EN
Publicado:	University of Modena and Reggio Emilia 2021
Materias:	Bombyx mori Nosema bombycis pebrine Loop-Mediated Isothermal Amplification NCBI Biology (General) QH301-705.5
Acceso en línea:	https://doaj.org/article/43c5ada4cf0e4d02a817f3d56e27fa86
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spelling	oai:doaj.org-article:43c5ada4cf0e4d02a817f3d56e27fa862021-12-02T18:20:53ZA Field-Friendly Loop-Mediated Isothermal Amplification (FF-LAMP) method for rapid detection of Nosema bombycis in silkworm, Bombyx mori1824-307Xhttps://doaj.org/article/43c5ada4cf0e4d02a817f3d56e27fa862021-06-01T00:00:00Zhttps://www.isj.unimore.it/index.php/ISJ/article/view/681https://doaj.org/toc/1824-307X Pebrine is a destructive disease that exhibits horizontal and vertical transmission and therefore it is the only mandatory quarantine item in sericulture. Here, a field-friendly loop-mediated isothermal amplification (FF-LAMP) method has been developed and validated for the rapid detection of Nosema bombycis, a causative agent of pebrine disease in silkworm, Bombyx mori. FF-LAMP primers were selected and designed for small ribosomal subunit gene and the assay was performed to detect the N. bombycis infection in silkworm. The FF-LAMP reaction was effective at 6 mM MgSO4, 1.4 mM dNTPs at 63 °C. The detection range of LAMP assay was found to be 101 dilutions of N. bombycis spores. Specificity of the primers was tested using DNA isolated from pebrine infected silkworm, pebrine free silkworm and pure N. bombycis by conventional PCR and FF-LAMP assay. Results revealed that the primers were specific to N. bombycis DNA. The FF-LAMP assay was validated in different basic silkworm seed farms with simultaneous microscopic examination of N. bombycis infection. This newly developed method is highly effective, specific, sensitive and rapid in detecting N. bombycis infection, eliminating the DNA purification steps and usage of sophisticated equipment. This method can be used in testing large number of samples making it field friendly method in sericulture industry. V SivaprasadL SatishG MallikarjunaN Chandrakanth A V Mary JosephaS M MoorthyUniversity of Modena and Reggio EmiliaarticleBombyx moriNosema bombycispebrineLoop-Mediated Isothermal AmplificationNCBIBiology (General)QH301-705.5ENInvertebrate Survival Journal, Vol 18, Iss 1 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Bombyx mori Nosema bombycis pebrine Loop-Mediated Isothermal Amplification NCBI Biology (General) QH301-705.5
spellingShingle	Bombyx mori Nosema bombycis pebrine Loop-Mediated Isothermal Amplification NCBI Biology (General) QH301-705.5 V Sivaprasad L Satish G Mallikarjuna N Chandrakanth A V Mary Josepha S M Moorthy A Field-Friendly Loop-Mediated Isothermal Amplification (FF-LAMP) method for rapid detection of Nosema bombycis in silkworm, Bombyx mori
description	Pebrine is a destructive disease that exhibits horizontal and vertical transmission and therefore it is the only mandatory quarantine item in sericulture. Here, a field-friendly loop-mediated isothermal amplification (FF-LAMP) method has been developed and validated for the rapid detection of Nosema bombycis, a causative agent of pebrine disease in silkworm, Bombyx mori. FF-LAMP primers were selected and designed for small ribosomal subunit gene and the assay was performed to detect the N. bombycis infection in silkworm. The FF-LAMP reaction was effective at 6 mM MgSO4, 1.4 mM dNTPs at 63 °C. The detection range of LAMP assay was found to be 101 dilutions of N. bombycis spores. Specificity of the primers was tested using DNA isolated from pebrine infected silkworm, pebrine free silkworm and pure N. bombycis by conventional PCR and FF-LAMP assay. Results revealed that the primers were specific to N. bombycis DNA. The FF-LAMP assay was validated in different basic silkworm seed farms with simultaneous microscopic examination of N. bombycis infection. This newly developed method is highly effective, specific, sensitive and rapid in detecting N. bombycis infection, eliminating the DNA purification steps and usage of sophisticated equipment. This method can be used in testing large number of samples making it field friendly method in sericulture industry.
format	article
author	V Sivaprasad L Satish G Mallikarjuna N Chandrakanth A V Mary Josepha S M Moorthy
author_facet	V Sivaprasad L Satish G Mallikarjuna N Chandrakanth A V Mary Josepha S M Moorthy
author_sort	V Sivaprasad
title	A Field-Friendly Loop-Mediated Isothermal Amplification (FF-LAMP) method for rapid detection of Nosema bombycis in silkworm, Bombyx mori
title_short	A Field-Friendly Loop-Mediated Isothermal Amplification (FF-LAMP) method for rapid detection of Nosema bombycis in silkworm, Bombyx mori
title_full	A Field-Friendly Loop-Mediated Isothermal Amplification (FF-LAMP) method for rapid detection of Nosema bombycis in silkworm, Bombyx mori
title_fullStr	A Field-Friendly Loop-Mediated Isothermal Amplification (FF-LAMP) method for rapid detection of Nosema bombycis in silkworm, Bombyx mori
title_full_unstemmed	A Field-Friendly Loop-Mediated Isothermal Amplification (FF-LAMP) method for rapid detection of Nosema bombycis in silkworm, Bombyx mori
title_sort	field-friendly loop-mediated isothermal amplification (ff-lamp) method for rapid detection of nosema bombycis in silkworm, bombyx mori
publisher	University of Modena and Reggio Emilia
publishDate	2021
url	https://doaj.org/article/43c5ada4cf0e4d02a817f3d56e27fa86
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A Field-Friendly Loop-Mediated Isothermal Amplification (FF-LAMP) method for rapid detection of Nosema bombycis in silkworm, Bombyx mori

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