Rapid detection of wheat yellow mosaic virus by reverse transcription loop-mediated isothermal amplification

Virology Journal

Cheng-Gui Han¹  Jia-Lin Yu¹  Da-Wei Li¹  Xiao-Jun Liu¹  Zong-Ying Zhang¹ 

[1] State Key Laboratory for Agrobiotechnology and Department of Plant Pathology, China Agricultural University, Beijing 100193, China

关键词: Virus detection;    RT-LAMP;    Wheat yellow mosaic virus;   

Others  :  1155422
DOI  :  10.1186/1743-422X-8-550

 received in 2011-09-22, accepted in 2011-12-20,  发布年份 2011

【 摘 要 】

For the detection of wheat yellow mosaic virus (WYMV), we established a reverse transcription loop-mediated isothermal amplification (RT-LAMP) method. Using Primer Explorer software, four sets of primers were designed and RT-LAMP assay reaction conditions were optimized. The RT-LAMP was performed at different times by four primer sets. Agarose gel analysis showed that WYMV could be detected after 30 min with the primer set III and after 45 min with the other three primer sets, both under the 80-min reaction time. RT-LAMP had the same results with the four primer sets, thus primer set III and 65°C for 80 min reaction were selected for virus detection. There was no significant different when avian myeloblastosis virus (AMV) and moloney murine leukemia virus (M-MLV) RT-LAMP with the four primer sets and M-MLV was chosen due to its relatively cheap price. The result on specificity showed that the assay could amplify WYMV specifically, and the sensitivity comparison showed that the RT-LAMP was 100 times more sensitive than conventional reverse-transcriptase-polymerase chain reaction (RT-PCR). Overall, RT-LAMP was found to be a simple, specific, sensitive, convenient and time-saving method for WYMV detection.

【 授权许可】

   
2011 Zhang et al; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

Figure 4.

Figure 5.

Figure 6.

Figure 7.

【 参考文献 】

• [1]Sawada E: Control of wheat yellow mosaic virus. J Plant Prot 1927, 14:444-449.
• [2]Kusume T, Tamada T, Hattori H, Tsuchiya T, Kubo K, Abe H, Namba S, Tsuchizaki T, Kishi K, Kashiwazaki S: Identification of a new wheat yellow mosaic virus strain with specific pathogenicity towards major wheat cultivars grown in Hokkaido. Ann Phytopathol Soc Jpn 1997, 63(2):107-109.
• [3]Wang M, Liu G, Lu X: A primary confirmation on wheat spindle streak mosaic virus occurring in China. Agric Sci Technol Sichuan 1980, 1:34-35.
• [4]Han C, Li D, Xing Y, Zhu K, Tian Z, Cai Z, Yu J, Liu Y: Wheat yellow mosaic virus widely occurring in wheat (Triticum aestivum) in China. Plant Dis 2000, 84(6):627-630.
• [5]Inouye T: Filamentous particles as the causal agent of yellow mosaic disease of wheat. Nogaku kenkyu 1969, 53:61-68.
• [6]Namba S, Kashiwazaki S, Lu X, Tamura M, Tsuchizaki T: Complete nucleotide sequence of wheat yellow mosaic bymovirus genomic RNAs. Arch Virol 1998, 143(4):631-643.
• [7]Li D, Yan L, Su N, Han C, Hou Z, Yu J, Liu Y: The nucleotide sequence of a Chinese isolate of wheat yellow mosaic virus and its comparison with a Japanese isolate. Arch Virol 1999, 144(11):2201-2206.
• [8]Hariri D, Delaunay T, Gomes L, Filleur S, Plovie C, Lapierre H: Comparison and differentiation of wheat yellow mosaic virus (WYMV), wheat spindle streak mosaic virus (WSSMV) and barley yellow mosaic virus (BaYMV) isolates using WYMV monoclonal antibodies. Eur J Plant Pathol 1996, 102(3):283-292.
• [9]Hariri D, Lapierre H, Filleur S, Plovie C, Delaunay T: Production and characterization of monoclonal antibodies to barley yellow mosaic virus and their use in detection of four bymoviruses. J Phytopathol 1996, 144(6):331-336.
• [10]Geng B, Han C, Zhai Y, Wang H, Yu J, Liu Y: Detection of wheat yellow mosaic virus by heterogeneous animal double-antibody sandwich ELISA. Virologica Sinica 2003, 18(1):76-78.
• [11]Clover G, Henry C: Detection and discrimination of wheat spindle streak mosaic virus and wheat yellow mosaic virus using multiplex RT-PCR. Eur J Plant Pathol 1999, 105(9):891-896.
• [12]Yue H, Wu Y, Li Y, Wei T, Hou W, Wu K: Simultaneous detection of three wheat virus BSMV, BYDV-PAV, WYMV and WBD phytoplasma by multiplex PCR. Sci Agri Sin 2008, 41(9):2663-2669.
• [13]Xing Y, Su N, Li D, Yu J, Liu Y: Over-expression of 72 kDa protein of wheat yellow mosaic virus in E. coli and preparation of its antiserum. Chin Sci Bull 2000, 45(6):525-528.
• [14]Dong J, He Z, Han C, Chen X, Zhang L, Liu W, Han Y, Wang J, Zhai Y, Yu J: Generation of transgenic wheat resistant to wheat yellow mosaic virus and identification of gene silence induced by virus infection. Chin Sci Bull 2002, 47(17):1446-1450.
• [15]Han C, Li D, Yu J, liu L, Shang Q, Liu Y: Preparation and application of specific antiserum against wheat yellow mosaic virus coat protein expressed in E. coli cells. J Agric Biotechnol 2002, 10(4):373-376.
• [16]Zhang Z, Xu J, Han C, Li D, Yu J: Detective and complete sequence analysis of wheat yellow mosaic virus from Zhumadian in Henan Province. Acta Agriculturae Boreali-Sinica 2010, 25(2):5-11.
• [17]Usugi T, Saito Y: Relationship between wheat yellow mosaic virus and wheat spindle streak mosaic virus. Ann Phytopathol Soc Jpn 1979, 45:397-400.
• [18]Notomi T, Okayama H, Masubuchi H, Yonekawa T, Watanabe K, Amino N, Hase T: Loop-mediated isothermal amplification of DNA. Nucleic Acids Res 2000, 28(12):e63.
• [19]Mori Y, Nagamine K, Tomita N, Notomi T: Detection of loop-mediated isothermal amplification reaction by turbidity derived from magnesium pyrophosphate formation. Biochem Biophys Res Commun 2001, 289(1):150-154.
• [20]Thai H, Le M, Vuong C, Parida M, Minekawa H, Notomi T, Hasebe F, Morita K: Development and evaluation of a novel loop-mediated isothermal amplification method for rapid detection of severe acute respiratory syndrome coronavirus. J Clin Microbiol 2004, 42(5):1956.
• [21]Arita M, Ling H, Yan D, Nishimura Y, Yoshida H, Wakita T, Shimizu H: Development of a reverse transcription-loop-mediated isothermal amplification (RT-LAMP) system for a highly sensitive detection of enterovirus in the stool samples of acute flaccid paralysis cases. BMC Infect Dis 2009, 9(1):208. BioMed Central Full Text
• [22]Jiang T, Liu J, Deng Y, Xu L, Li X, Han J, Cao R, Qin E: Development and evaluation of a reverse transcription-loop-mediated isothermal amplification assay for rapid detection of Enterovirus 71. J Clin Microbiol 2011, 49(3):870.
• [23]Nagdev K, Kashyap R, Parida M, Kapgate R, Purohit H, Taori G, Daginawala H: Loop-mediated isothermal amplification for rapid and reliable diagnosis of tuberculous menngitis. J Clin Microbiol 2011, 49(5):1861-1865.
• [24]Li S, Fang M, Zhou B, Ni H, Shen Q, Zhang H, Han Y, Yin J, Chang W, Xu G: Simultaneous detection and differentiation of dengue virus serotypes 1-4, Japanese encephalitis virus, and West Nile virus by a combined reverse-transcription loop-mediated isothermal amplification assay. Virol J 2011, 8(1):360. BioMed Central Full Text
• [25]Parida M, Shukla J, Sharma S, Santhosh S, Ravi V, Mani R, Thomas M, Khare S, Rai A, Ratho R: Development and evaluation of reverse transcription loop-mediated isothermal amplification assay for rapid and real-time detection of the swine-origin influenza A H1N1 virus. J Mol Diagnostics 2011, 13(1):100-107.
• [26]Fukuta S, Iida T, Mizukami Y, Ishida A, Ueda J, Kanbe M, Ishimoto Y: Detection of Japanese yam mosaic virus by RT-LAMP. Arch Virol 2003, 148(9):1713-1720.
• [27]Fukuta S, Kato S, Yoshida K, Mizukami Y, Ishida A, Ueda J, Kanbe M, Ishimoto Y: Detection of tomato yellow leaf curl virus by loop-mediated isothermal amplification reaction. J Virol Methods 2003, 112(1-2):35-40.
• [28]Fukuta S, Nimi Y, Oishi K, Yoshimura Y, Anai N, Hotta M, Fukaya M, Kato T, Oya T, Kambe M: Development of reverse transcription loop-mediated isothermal amplification (RT-LAMP) method for detection of two viruses and chrysanthemum stunt viroid. Ann Rep Kansai Plant Protect Soc 2005, 47:31-36.
• [29]Fukuta S, Ohishi K, Yoshida K, Mizukami Y, Ishida A, Kanbe M: Development of immunocapture reverse transcription loop-mediated isothermal amplification for the detection of tomato spotted wilt virus from chrysanthemum. J Virol Methods 2004, 121(1):49-55.
• [30]Le DT, Netsu O, Uehara-Ichiki T, Shimizu T, Choi IR, Omura T, Sasaya T: Molecular detection of nine rice viruses by a reverse-transcription loop-mediated isothermal amplification assay. J Virol Methods 2010, 170(1-2):90-93.
• [31]Nie X: Reverse transcription loop-mediated isothermal amplification of DNA for detection of Potato virus Y. Plant Dis 2005, 89(6):605-610.
• [32]Varga A, James D: Use of reverse transcription loop-mediated isothermal amplification for the detection of plum pox virus. J Virol Methods 2006, 138(1-2):184-190.
• [33]Boubourakas I, Fukuta S, Kyriakopoulou P: Sensitive and rapid detection of peach latent mosaic viroid by the reverse transcription loop-mediated isothermal amplification. J Virol Methods 2009, 160(1-2):63-68.
• [34]Tomita N, Mori Y, Kanda H, Notomi T: Loop-mediated isothermal amplification (LAMP) of gene sequences and simple visual detection of products. Nat Protoc 2008, 3(5):877-882.