Monoclonal Antibody-Based Serological Detection of Rice Stripe Mosaic Virus Infection in Rice Plants or Leafhoppers
Monoclonal Antibody-Based Serological Detection of Rice Stripe Mosaic Virus Infection in Rice Plants or Leafhoppers
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基于单克隆抗体的水稻或叶蝉水稻条纹花叶病毒感染血清学检测
DOI：
10.1007/s12250-019-00186-1
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发表时间：
2020-04-01
影响因子：
5.5
通讯作者：
Wu, Jianxiang
中科院分区：
文献类型：
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作者：
Guo, Liqian;Wu, Jiayu;Wu, Jianxiang
水稻条纹花叶病毒是近年来在中国南部发现的一种带状病毒，可造成水稻严重减产。为了建立RSMV流行病学研究的血清学方法，并建立RSMV的防治策略，我们首先从感染RSMV的水稻植株中提纯RSMV病毒粒子，然后将其作为免疫原制备4株RSMV特异性单抗(1D4、4A8、8E4和11F11)。利用这些单抗，我们建立了一种高度特异和灵敏的抗原包被平板酶联免疫吸附试验(ACP-ELISA)、一种斑点-ELISA和一种组织印迹-ELISA法，用于快速检测水稻或叶蝉的RSMV感染。结果表明，ACP-ELISA法和Dot-ELISA法分别在1：20,971,520(w/v，g/mL)稀释液和1：327,680(w/v，g/mL)稀释液中均能检测到RSMV。ACP-ELISA法和Dot-ELISA法均可用于检测以1：307,200和1：163,840(单株/亩L)稀释的桔小实蝇(Recilia Dorsalis)叶片匀浆中RSMV的侵染。对田间采集的水稻样品或带毒叶蝉的RSMV感染进行检测，结果表明，三种血清学方法均可获得与RT-PCR相同的结果(33个水稻样品中有19个样品和16个叶蝉样品中有5个样品呈RSMV阳性)。我们认为本研究获得的4株单抗具有高度的特异性和敏感性，这3种新的血清学方法对于检测水稻植株或叶蝉中的RSMV感染和制定病害防治策略是非常有用的。
Rice stripe mosaic virus (RSMV) is a rhabdovirus recently found in southern part of China and can cause severe reduction in rice production. To establish serological methods for RSMV epidemiological studies and to establish a control strategy for this virus, we first purified RSMV virions from infected rice plants and then used them as an immunogen to produce four RSMV-specific monoclonal antibodies (MAbs) (i.e.,1D4, 4A8, 8E4 and 11F11). With these MAbs, we have developed a highly specific and sensitive antigen-coated plate enzyme-linked immunosorbent assay (ACP-ELISA), a Dot-ELISA and a Tissue print-ELISA for rapid detections of RSMV infection in rice plants or in leafhoppers. Our results showed that RSMV can be readily detected in RSMV-infected rice plant tissue crude extracts diluted at 1:20,971,520 (w/v, g/mL) through ACP-ELISA or diluted at 1:327,680 (w/v, g/mL) through Dot-ELISA. Both ACP-ELISA and Dot-ELISA can also be used to detect RSMV infection in individual RSMV viruliferous leafhopper (Recilia dorsalis) homogenate diluted at 1:307,200 and 1:163,840 (individual leafhopper/mu L), respectively. Detection of RSMV infection in field-collected rice samples or in RSMV viruliferous leafhoppers indicated that the three serological methods can produce same results with that produced by RT-PCR (19 of the 33 rice samples and 5 of the 16 leafhoppers were RSMV-positive). We consider that the four MAbs produced in this study are very specific and sensitive, and the three new serological methods are very useful for detections of RSMV infection in rice plants or in leafhoppers and the establishment of the disease control strategies.