title: Detection of swine transmissible gastroenteritis coronavirus using loop-mediated isothermal amplification.
abstract: A conserved nucleic acid fragment of the nucleocapsid gene of Swine Transmissible Gastroenteritis Coronavirus (TGEV) was chosen as the target, six special primers were designed successfully. Loop-mediated isothermal amplification (LAMP) was developed to detect the TGEV by incubation at 60°C for 1 h and the product specificity was confirmed by HphI digestion. Standard curves with high accuracy for TGEV quantization was constructed by adding 1 × SYBR greenI in the LAMP reaction. The assay established in this study was found to detect only the TGEV and no cross-reaction with other viruses, demonstrating its high specificity. By using serial sample dilutions as templates, the detection limit of LAMP was about 10 pg RNA, 10 times more sensitive than that of PCR and could be comparable to the nest-PCR.
authors.affiliation: School of Life Science, Shanghai University, Shanghai 200444, China. chenqincc@yahoo.com.cn
authors.forename: Qin
authors.initials: Q
authors.lastname: Chen
authors.forename: Jian
authors.initials: J
authors.lastname: Li
authors.forename: Xue-En
authors.initials: XE
authors.lastname: Fang
authors.forename: Wei
authors.initials: W
authors.lastname: Xiong
mesh_terms: D000818:Animals; D052160:Benzothiazoles; D017124:Conserved Sequence; D017931:DNA Primers / Q000235:genetics; D003959:Diamines; D005761:Gastroenteritis, Transmissible, of Swine / Q000175:diagnosis* / Q000821:virology*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D019251:Nucleocapsid / Q000235:genetics; D009930:Organic Chemicals / Q000378:metabolism; D011804:Quinolines; D012680:Sensitivity and Specificity; D013194:Staining and Labeling / Q000379:methods; D013552:Swine; D013696:Temperature; D005760:Transmissible gastroenteritis virus / Q000302:isolation & purification*; D014773:Virology / Q000379:methods*
chemical_list: D052160:Benzothiazoles; D017931:DNA Primers; D003959:Diamines; D009930:Organic Chemicals; D011804:Quinolines; C098022:SYBR Green I
references.citation: J Vet Med B Infect Dis Vet Public Health. 2005 Apr;52(3):105-11
references.pmid: 15876221
references.citation: J Virol Methods. 2007 May;141(2):173-80
references.pmid: 17218021
references.citation: J Virol Methods. 2007 Dec;146(1-2):125-8
references.pmid: 17643501
references.citation: J Virol Methods. 2008 Jan;147(1):188-93
references.pmid: 17920701
references.citation: J Virol Methods. 2008 Jul;151(1):35-9
references.pmid: 18468699
references.citation: Nucleic Acids Res. 2000 Jun 15;28(12):E63
references.pmid: 10871386
references.citation: Arch Virol. 2006 Jun;151(6):1093-106
references.pmid: 16453084
references.citation: J Virol Methods. 2006 Jan;131(1):41-6
references.pmid: 16214229
references.citation: J Virol Methods. 1997 May;65(2):169-81
references.pmid: 9186940
references.citation: Arch Virol. 1979;60(2):161-6
references.pmid: 226036
references.citation: J Virol Methods. 1997 Jul;66(2):303-9
references.pmid: 9255741
references.citation: J Clin Microbiol. 2004 May;42(5):1956-61
references.pmid: 15131154