单头灰飞虱体内水稻条纹叶枯病毒的快速检测

Rapid Methods for the Detection of Rice Stripe Virus in Single Planthopper (Laodelphax striatellus)

利用RT-PCR检测感病植株和单头带毒灰飞虱,均扩增出水稻条纹叶枯病毒特有的一个长540 bp的片段,且人工饲毒虫的病毒含量高于自然感病稻田虫。以病毒S蛋白的多克隆抗血清,采用Western印迹技术从单虫体内检测到病毒抗原的2个专一性条带,大小分别为20.7和19.7 kDa。DIBA(dot immunobinding assay)检测法快速、简便,但专一性和灵敏度不及RT-PCR与Western印迹检测。对不同检测方法所得昆虫带毒率差异的原因进行了探讨。

RT-PCR was used to detect rice stripe virus （RSV） in single viruliferous planthoppers （Laodelphaxstriatellus） and in infected rice plants. A 540 bp product was specifically amplified, and its content was higher in artificially feeding insects than in naturally infected insects in the rice field. Western blot analysis revealed two protein bands, of 20.7 and 19.7 kDa, detectable from individual planthoppers by use of polyclonal antiserum against S protein （stripe disease-specific protein）. The DIBA （dot immunobinding assay） method is fast and simple to use in the detection of RSV antigen in single insects but has lower sensitivity and specificity than both RT-PCR and Western blot analysis. Possible explanations were discussed for the difference in proportion of RSV in L. striatellus found with use of these methods.