کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
3406662 | 1223582 | 2013 | 7 صفحه PDF | دانلود رایگان |
![عکس صفحه اول مقاله: Development of a loop-mediated isothermal amplification assay for rapid detection of bovine parvovirus Development of a loop-mediated isothermal amplification assay for rapid detection of bovine parvovirus](/preview/png/3406662.png)
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 °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.
► A LAMP assay was developed for rapid detection of bovine parvovirus.
► The sensitivity of LAMP assay was higher than PCR, detection limit of 9 copies/reaction.
► The LAMP was specific without any cross-reactivity with other related viruses.
► The addition of SYBR Green before the reaction could prevent better false positive reactions.
► This test was rapid and accurate.
Journal: Journal of Virological Methods - Volume 191, Issue 2, August 2013, Pages 155–161