NDLI: Development of a loop-mediated isothermal amplification assay for porcine circovirus type 2

Content Provider	Springer Nature Link
Author	Liu, Ye bing Zhang, Lei Xue, Qin hong Ning, Yi bao Zhang, Zhi gang
Copyright Year	2011
Abstract	In this study, the loop-mediated isothermal amplification (LAMP) method was used to develop a rapid and simple detection system for porcine circovirus type 2 (PCV2). According to the PCV2 sequences published in GenBank, multiple LAMP primers were designed targeting conserved sequences of PCV2. Using the DNA extracted from PCV2 isolates HUN-09 and SD-09 as the template, LAMP reactions in a PCV2 LAMP system was performed, the amplification products were detected by adding SYBR Green I and could be observed directly by the naked eye. The results showed highly-efficient and specific amplification in 30 min at 63°C with a LAMP real-time turbidimeter. Furthermore, PCV2 DNA templates, with a detection limit of 5.5×10$^{−5}$ ng of nucleic acid, indicated that this assay was highly sensitive. The results obtained with the naked eye after SYBR Green I staining were consistent with those detected by the real-time turbidimeter, showing the potential simplicity of interpretation of the assay results. The LAMP assay appeared to have greater accuracy than PCR and virus isolation for the analysis of 18 clinical samples. In addition it offers higher specificity and sensitivity, shorter reaction times and simpler procedures than the currently available methods of PCV2 detection. It is therefore a promising tool for the effective and efficient detection of PCV2.
Starting Page	214
Ending Page	220
Page Count	7
File Format	PDF
ISSN	16740769
Journal	Virologica Sinica
Volume Number	26
Issue Number	3
e-ISSN	1995820X
Language	English
Publisher	Springer-Verlag
Publisher Date	2011-06-12
Publisher Place	Berlin, Heidelberg
Access Restriction	Subscribed
Subject Keyword	Porcine circovirus type 2 (PCV2) Loop-mediated isothermal amplification (LAMP) Virus detection Virology Medical Microbiology Oncology Biochemistry Microbiology Microbial Genetics and Genomics
Content Type	Text
Resource Type	Article
Subject	Infectious Diseases Virology Immunology Molecular Medicine

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