Spodoptera litura Nucleopolyhedrovirus (SpltNPV) in Spodoptera litura Adults: Nested PCR Detection

doi:10.11924/j.issn.1000-6850.casb2022-0351

Abstract

Abstract:

The aim of this study is to establish a rapid and reliable method for molecular detecting of SpltNPV in Spodoptera litura adults, in order to overcome the defect of the conventional PCR technology. In this paper, the ecdysteroid UDP-glucosyltransferase (egt) gene of Spodoptera litura was used as the detection object. Nested PCR primers were designed based on the egt gene sequences, including a pair of outer primers egt-F/R and a pair of inner primers egt1-F/R. And the reliability of nested PCR detection was studied on Spodoptera litura adults. The results showed that the nested PCR method had strong specificity for SpltNPV DNA amplification, and the minimum limit for amplification detection of primer egt-F/R was 10^-7 μg/μL. The results also indicated that two rounds of conventional PCR amplification could not detect the SpltNPV by the primers of egt-F/R, and the nested PCR method could effectively amplify the target fragment of gene egt. In addition, the sensitivity of the nested PCR method was 2 orders of magnitude higher than that of the two rounds of the conventional PCR detection method. Therefore, the nested PCR method for detecting SpltNPV was established for Spodoptera litura adults in this study. Moreover, the method can provide technical support for the epidemiological study on the infection and transmission of the virus in Spodoptera litura adults.

Key words: Spodoptera litura Nucleopolyhedrovirus (SpltNPV), adult, egt, nested PCR, detection

PU Xiaoming, DENG Haibin, LU Zheng, ZHANG Jingxin, SUN Dayuan, SHEN Huifang, LIU Pingping, LIN Birun, YANG Qiyun. Spodoptera litura Nucleopolyhedrovirus (SpltNPV) in Spodoptera litura Adults: Nested PCR Detection[J]. Chinese Agricultural Science Bulletin, 2023, 39(9): 128-133.

Figures/Tables 5

References 27

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昆虫病毒	egt基因号(NCBI)	egt基因大小/bp	基因序列同源率/%
斜纹夜蛾核型多角体病毒	EU374639.1	1569	/
苜蓿银纹夜蛾核型多角体病毒	M22619.1	2792	22.83
甘蓝夜蛾核型多角体病毒	U41999.1	1781	37.30
棉铃虫核型多角体病毒	AF000009.1	2111	30.93
舞毒蛾核型多角体病毒	U04321.1	1688	38.15
油桐尺蠖核型多角体病毒	U61154.1	1980	32.80
黎豆夜蛾核型多角体病毒	AF313417.1	1635	37.11

昆虫病毒	egt基因号(NCBI)	egt基因大小/bp	基因序列同源率/%
斜纹夜蛾核型多角体病毒	EU374639.1	1569	/
苜蓿银纹夜蛾核型多角体病毒	M22619.1	2792	22.83
甘蓝夜蛾核型多角体病毒	U41999.1	1781	37.30
棉铃虫核型多角体病毒	AF000009.1	2111	30.93
舞毒蛾核型多角体病毒	U04321.1	1688	38.15
油桐尺蠖核型多角体病毒	U61154.1	1980	32.80
黎豆夜蛾核型多角体病毒	AF313417.1	1635	37.11