Kajiwara Hideyuki, Murakami Ritsuko
National Agriculture and Food Research Organization, Kannondai 3-1-1, Tsukuba, Ibaraki 305-8517, Japan.
National Agriculture and Food Research Organization, Kannondai 3-1-1, Tsukuba, Ibaraki 305-8517, Japan.
Anal Biochem. 2017 Dec 15;539:45-47. doi: 10.1016/j.ab.2017.10.003. Epub 2017 Oct 6.
There is a need for rapid and less expensive methods to identify RNA viruses, including luteoviruses, for practical use in agriculture and quarantine. The mass spectrometric cleaved amplified polymorphic sequence (MS-CAPS) method, which detects enzymatically cleaved amplicons by matrix-assisted laser desorption/ionization mass spectrometry, was herein used together with a short RT-PCR to detect luteovirus in only 90 min. In addition, the matrixes 2',4',6'-trihydroxyacetophene and 3-hydroxypicolinic acid were compared for their effectiveness in the analysis of short single-stranded biotinylated DNA obtained by a MS-CAPS reaction.
需要快速且成本较低的方法来鉴定包括黄矮病毒属病毒在内的RNA病毒，以便在农业和检疫中实际应用。质谱裂解扩增多态性序列（MS-CAPS）方法通过基质辅助激光解吸/电离质谱检测酶切扩增子，本文将其与短RT-PCR一起使用，仅需90分钟即可检测黄矮病毒属病毒。此外，还比较了2',4',6'-三羟基苯乙酮和3-羟基吡啶甲酸这两种基质在分析通过MS-CAPS反应获得的短单链生物素化DNA中的有效性。
