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. 2015 Sep;26(3):203-6.
doi: 10.1007/s13337-015-0265-9. Epub 2015 Jul 28.

Detection of Aleutian disease virus by loop-mediated isothermal amplification

Zhuo Zhang  1 , Bin Wang  2 , Shouping Hu  2 , Jiaoer Zhang  2 , Xijun He  2 , Shimin Zheng  3
Affiliations

Detection of Aleutian disease virus by loop-mediated isothermal amplification

Zhuo Zhang et al. Virusdisease. 2015 Sep.

Abstract

In this study, a loop-mediated isothermal amplification (LAMP) assay was developed and optimized for the detection of Aleutian disease virus (ADV) in minks. The amplification could be completed within 45 min under isothermal condition by employing a set of six ADV genome-specific primers. The amplification results could be visualized directly with the naked eye by using fluorescent dye. Comparative experiments showed that the LAMP assay is superior to conventional polymerase chain reaction for the detection of both experimental and field samples. Results of current study indicated that the LAMP assay is a rapid and reliable technique for routine diagnosis of ADV infection in minks.

Keywords: Aleutian disease virus; Diagnosis; Loop-mediated isothermal amplification.

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Figures

Fig. 1
Fig. 1
Oligonucleotide sequences of LAMP primers
Fig. 2
Fig. 2
LAMP products generated at different time intervals. Samples were removed one by one at 15 min interval to test the amplification of template DNA. NC negative control
Fig. 3
Fig. 3
Comparative sensitivity of LAMP and PCR for the detection of ADV genome. Template DNA was 10 times serially diluted (from 3.3 × 106 to 3.3 × 100 copies). LAMP and PCR results were determined through agarose gel analysis (a and c). The positive LAMP reactions can also be inspected directly by naked eye (b). NC negative control
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