Rapid and sensitive detection of shrimp yellow head virus using loop-mediated isothermal amplification and a colorogenic nanogold hybridization probe

被引:45
作者
Jaroenram, Wansadaj [1 ,2 ]
Arunrut, Narong [1 ,3 ]
Kiatpathomchai, Wansika [1 ,3 ]
机构
[1] Mahidol Univ, Fac Sci, CENTEX Shrimp, Bangkok 10400, Thailand
[2] James Cook Univ, Sch Vet & Biomed Sci, Townsville, Qld 4811, Australia
[3] Natl Sci & Technol Dev Agcy, Natl Ctr Genet Engn & Biotechnol BIOTEC, Pathum Thani 12120, Thailand
关键词
YHV; RT-LAMP; Gold nanoparticle; Colorogenic nanogold hybridization; LATERAL FLOW DIPSTICK; UNMODIFIED GOLD NANOPARTICLES; NUCLEIC-ACID TARGETS; TAURA SYNDROME VIRUS; SPOT SYNDROME VIRUS; OPTICAL-PROPERTIES; PENAEUS-MONODON; DNA; TRANSITION; MONOLAYERS;
D O I
10.1016/j.jviromet.2012.08.013
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Salt-induced self-aggregation of gold nanoparticles (AuNP) carrying unisense ssDNA probes can be prevented specifically by complementary DNA. Loop-mediated isothermal amplification (LAMP) can amplify DNA rapidly. Here, the two techniques were combined to detect yellow head virus (YHV). The LAMP-AuNP method required 60 min for LAMP and 5 min for hybridization of LAMP products to an AuNP-labeled ssDNA probe followed by salt induced probe-particle aggregation to visualize color development. The detection sensitivity of the method was comparable to that of the commercial IQ2000 (TM) nested RT-PCR but only required similar to 65 min to produce a result, and did not cross-detect other shrimp viruses. As the LAMP-AuNP protocol only requires a heating block, it offers opportunities for rapid detection of YHV. (C) 2012 Elsevier B.V. All rights reserved.
引用
收藏
页码:36 / 42
页数:7
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