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Development of loop-mediated isothermal amplification assay for specific and rapid detection of camelpox virus in clinical samples.

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Title Development of loop-mediated isothermal amplification assay for specific and rapid detection of camelpox virus in clinical samples.
 
Creator Venkatesan G, Bhanuprakash V, Balamurugan V, Singh RK, Pandey AB.
 
Subject Camelpox Virus, LAMP, Detection
 
Description In this study, development of loop-mediated isothermal amplification (LAMP) assay based on ankyrin repeat protein gene (C18L) for specific and rapid detection of camelpox virus (CMLV) was carried out. The assay was optimized using viral genomic DNA (gDNA) extracted from density gradient purified CMLV and standard control recombinant DNA plasmid containing the target, which resulted in reliable amplification at 62°C for 60 min. The amplified LAMP product was identified by agarose gel electrophoresis and subsequent direct visualization under UV light or observation by naked-eye for the presence of turbidity and color change following the addition of SYBR Green I dye and hydroxy naphthol blue (HNB). The analytical specificity of LAMP and conventional PCR assays was evaluated using other related poxviruses namely buffalopox, goatpox, sheeppox, and orf viruses, which revealed only a specific amplification of CMLV. The LAMP assay was 10-fold more sensitive than the conventional PCR. Further, the assay was evaluated with DNA extracted from the cell culture isolates of CMLV (n=11) and clinical samples (n=23). These results proved that the developed LAMP is a simple, specific, sensitive, rapid and economical diagnostic tool for detection of CMLV from clinical materials.
 
Date 2023-05-12T10:21:01Z
2023-05-12T10:21:01Z
2012-07-01
 
Type Research Paper
 
Identifier Not Available
http://krishi.icar.gov.in/jspui/handle/123456789/77235
 
Language English
 
Publisher Elsevier