Loop-mediated isothermal amplification establishment for detection of pseudorabies virus

被引：77

作者：

En, Fang-Xue ^{[1
]}

Wei, Xiong ^{[2
]}

Jian, Li ^{[2
]}

Qin, Chen ^{[1
]}

机构：

[1] Shanghai Univ, Sch Life Sci, Shanghai 200444, Peoples R China

[2] Shanghai Entry Exit Inspect & Quarantine Bur, Shanghai 200135, Peoples R China

来源：

JOURNAL OF VIROLOGICAL METHODS | 2008年 / 151卷 / 01期

关键词：

LAMP; PRV; detection;

D O I：

10.1016/j.jviromet.2008.03.028

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

A rapid, convenient and reliable pseudorabies virus (PRV) detection system was developed by using the loop-mediated isothermal amplification (LAMP) method. Six special primers were designed successfully based on the PRV DNA-binding protein (DBP) gene. The assay was optimized to amplify PRV DNA by incubation at 63 degrees C for 1 h. The LAMP products had a ladder-like pattern of bands from 188 bp when electrophoresed on an agarose gel and its specificity was confirmed by digestion with Hinc II enzyme. Two naked-eye detection methods were developed for use in the field. The detection limit of the LAMP assay was found to be 10 fg DNA sample which was 100-1000-fold higher than that of PCR. By using DNA (or cDNA) samples extracted from three different PRV strains and six other viruses known to be related genetically to PRV or to cause similar clinical signals in pig, the system was identified to amplify only the PRV DNA. A comparison between the LAMP and PCR assay using five clinical samples showed good correlation. (C) 2008 Elsevier B.V. All rights reserved.

引用

页码：35 / 39

页数：5

共 16 条

[1] Aujeszky's disease (pseudorabies) virus detection in cerebrospinal fluid in experimentally infected pigs [J].

Balasch, M ;

Pujols, J ;

Segalés, J ;

Pumarola, M .

VETERINARY MICROBIOLOGY, 1998, 60 (2-4) :99-106

[2] A one-step reverse transcriptase loop-mediated isothermal amplification assay for simple and rapid detection of swine vesicular disease virus [J].

Blomstrom, Anne-Lie ;

Hakhverdyan, Mikhayil ;

Reid, Scott M. ;

Dukes, Juliet P. ;

King, Donald P. ;

Belak, Sandor ;

Berg, Mikael .

JOURNAL OF VIROLOGICAL METHODS, 2008, 147 (01) :188-193

[3]

CHEN HCH, 2002, DIAGNOSTIC TECHNIQUE

[4] IMMUNOPEROXIDASE STUDY OF AUJESZKYS DISEASE IN PIGS [J].

DUCATELLE, R ;

COUSSEMENT, W ;

HOORENS, J .

RESEARCH IN VETERINARY SCIENCE, 1982, 32 (03) :294-302

[5] Novel reverse transcription loop-mediated isothermal amplification for rapid detection of foot-and-mouth disease virus [J].

Dukes, J. P. ;

King, D. P. ;

Alexandersen, S. .

ARCHIVES OF VIROLOGY, 2006, 151 (06) :1093-1106

[6] Rapid diagnosis of H5N1 avian influenza virus infection by newly developed influenza H5 hemagglutinin gene-specific loop-mediated isothermal amplification method [J].

Imai, Masaki ;

Ninomiya, Ai ;

Minekawa, Harumi ;

Notomi, Tsugunori ;

Ishizaki, Toru ;

Tu, Phan Van ;

Tien, Nguyen Thi Kim ;

Tashiro, Masato ;

Odagiri, Takato .

JOURNAL OF VIROLOGICAL METHODS, 2007, 141 (02) :173-180

[7] Rapid and sensitive detection of Taura syndrome virus by reverse transcription loop-mediated isothermal amplification [J].

Kiatpathomchai, Wansika ;

Jareonram, Wansadaj ;

Jitrapakdee, Sarawut ;

Flegel, T. W. .

JOURNAL OF VIROLOGICAL METHODS, 2007, 146 (1-2) :125-128

[8] Multiplex PCR for the simultaneous detection of pseudorabies virus, porcine cytomegalovirus, and porcine circovirus in pigs [J].

Lee, Chul-Seung ;

Moon, Hyoung-Joon ;

Yang, Jeong-Sun ;

Park, Seong-Jun ;

Song, Dae-Sub ;

Kang, Bo-Kyu ;

Park, Bong-Kyun .

JOURNAL OF VIROLOGICAL METHODS, 2007, 139 (01) :39-43

[9]

LI QY, 2005, LETT BIOTECH, V16, P647

[10] Detection of loop-mediated isothermal amplification reaction by turbidity derived from magnesium pyrophosphate formation [J].

Mori, Y ;

Nagamine, K ;

Tomita, N ;

Notomi, T .

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 2001, 289 (01) :150-154

← 1 2 →