Rapid, Sensitive and Simultaneous Detection of Two Wheat RNA Viruses Using Reverse Transcription Recombinase Polymerase Amplification (RT-RPA)

被引:7
|
作者
Chen, Zhiqing [1 ]
Zhang, Tianye [1 ,2 ]
Lei, Jiajia [1 ]
Wang, Ziqiong [1 ,2 ]
Liu, Peng [1 ]
Zhong, Kaili [1 ]
Chen, Jianping [1 ]
Liu, Jiaqian [1 ]
机构
[1] Ningbo Univ, Inst Plant Virol, State Key Lab Qual & Safety Agroprod, Ningbo 315211, Peoples R China
[2] Nanjing Agr Univ, Coll Plant Protect, Nanjing 210095, Peoples R China
来源
LIFE-BASEL | 2022年 / 12卷 / 12期
基金
中国国家自然科学基金;
关键词
wheat yellow mosaic virus; Chinese wheat mosaic virus; viral detection; RT-RPA; YELLOW-MOSAIC-VIRUS; DWARF VIRUS; RESISTANCE; SEQUENCE;
D O I
10.3390/life12121952
中图分类号
Q [生物科学];
学科分类号
07 ; 0710 ; 09 ;
摘要
In China, wheat yellow mosaic disease is mostly caused by wheat yellow mosaic virus (WYMV) and Chinese wheat mosaic virus (CWMV). If wheat is co-infected with these two viruses, it can cause severe yellow mosaic symptoms and yield losses. Early detection of viruses is crucial for preventing disease in the field. In this study, we optimized a sensitive, specific reverse transcription recombinase polymerase amplification (RT-RPA) detection method for two viruses, WYMV and CWMV. Two sets of primers were designed based on the capsid protein (CP)-encoding genes of the two viruses, and the reaction conditions were determined. The RT-RPA method, which amplified the target amplicon by a handheld reaction mixture for 20 min, was more sensitive than PCR-CP in the detection of WYMV. Finally, the RT-RPA method was performed on 110 randomly selected field samples, demonstrating its applicability to samples from different regions and specificity for co-infected samples. This study not only describes an improved method for detecting WYMV and CWMV using RT-RPA but also demonstrates the potential of this method, which could be applied under field conditions.
引用
收藏
页数:14
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