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一种用于检测柑橘碎叶病毒的实时逆转录定量聚合酶链反应检测方法。

A real-time RT-qPCR assay for the detection of Citrus tatter leaf virus.

作者信息

Cowell S J, Harper S J, Dawson W O

机构信息

Citrus Research and Education Center, University of Florida, Lake Alfred, FL, USA.

Citrus Research and Education Center, University of Florida, Lake Alfred, FL, USA.

出版信息

J Virol Methods. 2017 Jun;244:29-31. doi: 10.1016/j.jviromet.2017.03.004. Epub 2017 Mar 6.

Abstract

Citrus tatter leaf virus (CLTV) is globally distributed wherever citrus is grown, and, given the extensive use of CTLV sensitive rootstock, has the potential to be a significant threat to the citrus industry. In order to facilitate fast and reliable detection of this virus, we have developed a CTLV-specific real-time RT-qPCR assay. The optimized assay was found to be more reliable and sensitive compared to ELISA and end-point RT-PCR, detecting CTLV in up to 70% more plants. The real-time RT-qPCR is also specific, as it did not cross-react with the closely related Apple stem grooving virus or with the host itself; robust, being able to detect CTLV in young and mature host tissue types; and rapid.

摘要

柑橘碎叶病毒(CLTV)在全球所有种植柑橘的地方均有分布,鉴于对CLTV敏感的砧木被广泛使用,它有可能对柑橘产业构成重大威胁。为了便于快速、可靠地检测这种病毒,我们开发了一种针对CLTV的实时逆转录定量聚合酶链反应(RT-qPCR)检测方法。结果发现,与酶联免疫吸附测定(ELISA)和终点逆转录聚合酶链反应(end-point RT-PCR)相比,优化后的检测方法更可靠、更灵敏,能够检测出多70%的感染CLTV的植株。实时RT-qPCR也具有特异性,因为它不会与密切相关的苹果茎沟病毒或宿主本身发生交叉反应;具有稳健性,能够在年轻和成熟的宿主组织类型中检测到CLTV;而且检测速度快。

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