Nie X, Singh R P
Agriculture and Agri-Food Canada, Potato Research Centre, PO Box 20280, Fredericton, New Brunswick, Canada.
J Virol Methods. 2000 May;86(2):179-85. doi: 10.1016/s0166-0934(00)00140-3.
A novel usage of multiplex reverse transcription polymerase chain reaction (m-RT-PCR) for simultaneous detection of multiple viruses is reported. By use of an oligo(dT), as a common primer, nearly full-length cDNAs can be synthesized. Furthermore, combining an oligo(dT) primer with a specific antisense primer can be used to simultaneously prime reverse transcription of both polyadenylated and non-polyadenylated RNAs. Four viral genera including five potato viruses [(carlavirus (PVS), polerovirus (PLRV), potexvirus (PVX), potyvirus (PVA and PVY))] and a viroid genus including a viroid genome (pospiviroid (PSTVd)) were used to develop various formats of m-RT-PCR. In artificially created viral RNA mixtures, all six RNA pathogens were detected successfully by uniplex- and m-RT-PCR. In naturally infected field grown tubers, m-RT-PCR detected infection of two to three viruses, which were present in the tubers.
报道了多重逆转录聚合酶链反应（m-RT-PCR）用于同时检测多种病毒的新用途。通过使用寡聚（dT）作为通用引物，可以合成几乎全长的cDNA。此外，将寡聚（dT）引物与特异性反义引物结合，可用于同时引发聚腺苷酸化和非聚腺苷酸化RNA的逆转录。使用包括五种马铃薯病毒的四个病毒属[（香石竹潜隐病毒（PVS）、马铃薯卷叶病毒（PLRV）、马铃薯X病毒（PVX）、马铃薯Y病毒（PVA和PVY））]和包括一种类病毒基因组的一个类病毒属[马铃薯纺锤块茎类病毒（PSTVd）]来开发各种形式的m-RT-PCR。在人工构建的病毒RNA混合物中，通过单重和m-RT-PCR成功检测到所有六种RNA病原体。在自然感染的田间种植块茎中，m-RT-PCR检测到块茎中存在的两到三种病毒感染。
