期刊
JOURNAL OF VIROLOGICAL METHODS
卷 191, 期 2, 页码 155-161出版社
ELSEVIER
DOI: 10.1016/j.jviromet.2012.05.002
关键词
Bovine parvovirus; LAMP; Detection
资金
- National Scientific and Technical Supporting Programs of the Ministry of Science and Technology of China [2009BADB4B01]
- Foundation of Doctor Research of Northeast Agricultural University [2009RC03]
A loop-mediated isothermal amplification (LAMP) assay was developed for detection of bovine parvovirus (BPV) DNA. Four primers were designed to recognize six distinct regions on the target DNA based on a highly conserved sequence in the VP2 region of the BPV genome. The optimized LAMP reaction conditions were 8 mM Mg2+, 1.2 mM betaine, and an incubation at 63 degrees C for 45 min. After amplification the products were detected either by observing a ladder pattern following gel electrophoresis, observation of turbidity, or a color change with the addition of SYBR Green I to the reaction tube. The detection limit of the LAMP assay was 9 copies of BPV-DNA and was 100 times more sensitive than conventional PCR. A ladder pattern of bands after gel electrophoresis was observed for only BPV isolates and showed that the BPV LAMP assay was highly specific without any cross-reactivity with other related viruses. The LAMP assay was evaluated further using 59 field samples and the results were comparable to conventional PCR. The LAMP assay is a simple, rapid and economic detection method; it can provide a useful technique suitable for detection of BPV infection in both field conditions and laboratory settings. (C) 2012 Elsevier B.V. All rights reserved.
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