IVT反应副产物，dsRNA，检测方案

双链(ds)RNA的形成是病毒感染的标志，对于诱导先天免疫至关重要。dsRNA还参与基因沉默，并在体外转录基因合成过程中作为副产物产生。抗dsRNA单克隆抗体是在细胞培养和组织中检测dsRNA的有效工具，主要应用于如下领域：  

1.用于表征和检测具有dsRNA基因组或中间体的病毒（包括SARS，丙型肝炎，登革热或西尼罗河病毒）。

2.作为诊断工具，用于确定未知病原体是病毒还是细菌来源

3.用于体外转录(m)RNA制备的质量控制 

dsRNA抗体精选与推荐：

Fig.1：A）使用抗双链RNA单克隆抗体J2在SARS-CoV感染的Vero细胞中检测dsRNA的免疫荧光。 1：未感染的细胞。 标尺：20微米（根据[4]进行修改）。 B）使用抗双链RNA单克隆抗体J2在SARS-Cov感染的Vero E6细胞的双膜囊泡（DMVs）中进行免疫电子显微镜检测（根据[5]进行修改）。 C）使用抗双链RNA单克隆抗体J2在柯萨奇病毒感染的新生小鼠FFPE组织中检测dsRNA。 1：小鼠心脏，2：褐色脂肪，3：胰腺，4：唾液腺，5：中枢神经系统神经节，6：未感染组织。  

《Cell文章推荐》：

SND1 binds SARS-CoV-2 negative-sense RNA and promotes viral RNA synthesis through NSP9，Cell  |  2023 Oct 26

Article Snippet

".. washed once with ice-cold PBS, followed by two washes with PBS at room temperature.. For combining immunostaining with HCR-based detection, we used anti-J2 antibody (Jena Bioscience, RNT-SCI-10010200, 1:500) or anti-SND1 antibody (Proteintech, 60265-1- Ig, 1:500).. Incubation with the primary antibody was performed for 1 h at room temperature.Incubation with .."

Figure Legend

"... (B) Representative images of IF staining for dsRNA (J2 antibody) using HCR detection in SARS-CoV-2 infected"   

更多历史参考文献： 

 [1] Sch?nborn et al. (1991) Monoclonal antibodies to double-stranded RNA as probes of RNA structure in crude nucleic acid extracts. Nucleic Acids Res.19: 2993. 

 [2] Lukacs (1994) Detection of virus infection in plants and differentiation between coexisting viruses by monoclonal antibodies to double-stranded RNA. J. Virol. Methods47: 255. 

 [3] Lukacs (1997) Detection of sense:antisense duplexes by structure-specific anti-RNA antibodies. In: Antisense Technology. A Practical Approach, C. Lichtenstein and W. Nellen (eds), pp. 281-295. IRL Press, Oxford 

 [4] Weber et al. (2006) Double-Stranded RNA is produced by positive-strand RNA viruses and DNA viruses but not in detectable amounts by negative-strand RNA viruses. Journal of Virology 80(10): 5059. 

 [5] Knoops et al. (2008) SARS-Coronavirus replication is supported by a reticulovesicular network of modified endoplasmic reticulum. PLOS Biology 6(9): e226.

 [6] Richardson et al. (2010) Use of antisera directed against dsRNA to detect viral infections in formalin-fixed paraffin-embedded tissue. Journal of Clinical Virology 49: 180. 

 [7] Karikó et al. (2011) Generating the optimal mRNA for therapy: HPLC purification eliminates immune activation and improves translation of nucleoside-modified, protein-encoding mRNA. Nucleic Acids Research 39(21): e142.