用于传染性支气管炎病毒核酸检测的N蛋白基因装甲RNA质控品的制备

doi:10.11843/j.issn.0366-6964.2025.11.034

Abstract

Abstract:

The objective of this study is to develop a quality-controled product that can be used in the RNA detection of infectious bronchitis virus (IBV). The relatively conserved N protein gene of IBV was selected as the target sequence, and it was cloned into the vector pET-28a(+) with the MS2 phage capsid protein gene and packaging site sequence, and the RNA containing the N protein gene sequence of IBV was obtained by induction expression and purification, and its physical characteristics were observed by electron microscopy, and the copy number was accurately quantified by digital PCR. The real-time RT-PCR method introduced in the national standard GB/T23197—2022 was used to verify the detection of purified armor RNA, and the RNase enzyme resistance and storage stability were verified as well. The armored RNA containing IBV N gene was successfully constructed, which was resistant to the degradation of RNase enzyme and could be stably stored at 37℃ for above 15 days. The developed armor RNA has the potential to become a positive control product for IBV, which can realize the quality control of the whole process of RNA virus detection.

Key words: avian infectious bronchitis, protein N, quality control sample, nucleic acid detection, armored RNA

CLC Number:

S852.659.2

XIONG Yaoyu, LIU Dan, GAO Jianshuai, LI Huitong, ZHANG Boyuan, DING Jiabo, JIANG Hui, FAN Xuezheng, SHEN Qingchun. Preparation of N-protein Gene Armor RNA Control for Infectious Bronchitis Virus Nucleic Acid Detection[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(11): 5770-5777.

Figures/Tables 7

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References 27

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差异源 Source of difference	平方和 SS	自由度 df	均方 MS	F值 F-value	P值 P-value	F临界值 F crit
组间Between groups	0.009 12	2	0.004 56	0.670 26	0.529 726	3.885 294
组内Within groups	0.081 64	12	0.006 803
总计Total	0.090 76	14	/	/	/	/

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Preparation of N-protein Gene Armor RNA Control for Infectious Bronchitis Virus Nucleic Acid Detection

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