African Journal of
Microbiology Research

  • Abbreviation: Afr. J. Microbiol. Res.
  • Language: English
  • ISSN: 1996-0808
  • DOI: 10.5897/AJMR
  • Start Year: 2007
  • Published Articles: 5233

Full Length Research Paper

One-hour loop-mediated isothermal amplification assay for the detection of quarantinable toxigenic Fusarium garmanirum

Hassan Almoammar1,2*, Ali H. Bahkali2,3 and Kamel A. Abd-Elsalam4
1King Abdulaziz City for Science and Technology (KACST), P. O. Box 6086, Riyadh 11442, Kingdom of Saudi Arabia. 2Botany and Microbiology Department, Faculty of Science, King Saud University, Riyadh, Saudi Arabia. 3Center of Excellence in Biotechnology Research, King Saud University, P.O. Box 2455, Riyadh 11451, Saudi Arabia. 4Unit of Excellence in N ano-Molecular Plant Pathology Research (ARC), Egypt.
Email: [email protected]

  •  Accepted: 25 March 2013
  •  Published: 02 April 2013

Abstract

Fusarium head blight (FHB) caused by several Fusarium species is one of the most serious diseases affecting wheat throughout the world. The in vitro production of the toxins deoxynivalenol, zearalenone fumonisin, T-2, and HT-2 was quantitvely evaluated in 8 different isolates of Fusarium species collected from feed samples. It was possible to detect zearalenone and the other mycotoxins in 100% and 50% of the isolates, respectively. In the present study,  loop-mediated  isothermal  amplification  method  (LAMP) was designed for diagnosing  Fusarium garmanirum  infections  and  testing  against  feed samples, infested samples and pure cultures. The  LAMP  amplicon  was  directly  visualized  in  the reaction  tubes  by  the  naked  eye  following  the  addition  of calcein fluorescence. The LAMP products appeared as DNA marker pattern, with many bands of different sizes from 145 base pairs up to the loading well. Loop-LAMP procedure was used to detect genomic DNA of F. graminearum in fungal pure culture and in contaminated feed samples. In the future, this assay will support plant quarantine programs in Saudi Arabia and Gulf Cooperation Council states, to prevent the introduction of foreign FHB species.

 

Key words: LAMP-PCR, Fusarium head blight, feed samples.