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  • 标题:New closed tube loop mediated isothermal amplification assay for prevention of product cross-contamination
  • 本地全文:下载
  • 作者:K. Karthik ; Rajesh Rathore ; Prasad Thomas
  • 期刊名称:MethodsX
  • 印刷版ISSN:2215-0161
  • 电子版ISSN:2215-0161
  • 出版年度:2014
  • 卷号:1
  • 页码:137-143
  • DOI:10.1016/j.mex.2014.08.009
  • 语种:English
  • 出版社:Elsevier
  • 摘要:Graphical abstractAbstractLoop mediated isothermal amplification (LAMP) assay, a promising diagnostic test, has been developed for detection of different pathogens of human as well as animals. Various positive points support its use as a field level test but the major problem is product cross contamination leading to false positive results. Different methods were adopted by various researchers to control this false positive amplification due to cross contamination but all have their own advantages and disadvantages. A new closed tube LAMP assay based on agar dye capsule was developed in the present study and this technique has some advantages over the other closed tube technique.•Agar at the concentration of 1.5% was used to sandwich SYBR green dye I with the aid of intradermal syringe. This agar dye capsule was placed over the LAMP reaction mixture before it was amplified.•To eliminate the hazardous nature of Ultra Violet (UV) light during result visualization of LAMP products, the present study demonstrates the use of Light Emitting Diode (LED) lights for result visualization.•LAMP was carried out forBrucellaspecies detection using this modified techniques yielding good results without any cross contamination and LED showed similar fluorescence compared to UV.
  • 关键词:LAMP;Brucella;Contamination;Agar dye capsule;LED;SYBR green
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