猫疱疹病毒1型RPA-Cas12a-LFD检测方法的建立及初步应用

doi:10.11843/j.issn.0366-6964.2022.05.032

Abstract

Abstract: Feline herpesvirus-1 (FHV-1) is double-stranded DNA enveloped virus of Varicellovirus, is one of major causative pathogen for feline upper respiratory tract infection. Based on recombinase polymerase amplification (RPA), Cas12a trans-cleavage reaction and lateral flow dipstick (LFD), a rapid visulization method for FHV-1 detection (RPA-Cas12a-LFD) was developed and validated. RPA amplified primers and crRNA synthesized primers were designed by targeting the FHV-1 TK gene, and then validation of RPA-Cas12a-LFD reaction system, sensitivity and specificity tests, and consistensy analysis of TB Green qPCR and RPA-Cas12a-LFD for FHV-1 detection were performed respectively. The results showed that the RPA-Cas12a-LFD method could specifically detect FHV-1 without cross-reaction with other associated pathogens, with advantages of high sensitivity (detection limit was 2.35×10^-1copies·μL^-1), short detection time and visual readout. The detection rate of FHV-1 in 20 clinical samples (35%, 7/20) was higher than TB Green qPCR (25%, 5/20) and the coincidence rate of 5 positive samples was 100%. In conclusion, RPA-Cas12a-LFD method was of good specificity and ultrasensitivity for FHV-1 detection without special equipment, and could be a promising and reliable tool for rapid on-site diagnosis.

Key words: feline herpesvirus-1, RPA-Cas12a-LFD, nucleic acid detection, application

CLC Number:

S852.659.6

HUANG Jian, LIU Yunjia, YANG Xiaonong, LI Yan. RPA-Cas12a-LFD Based Nucleic Acid Detection for Feline Herpesvirus-1 and Preliminary Application[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(5): 1638-1643.

References 17

[1]	LEWIN A C, KOLB A W, MCLELLAN G J, et al. Genomic, recombinational and phylogenetic characterization of global feline herpesvirus 1 isolates[J]. Virology, 2018, 518:385-397.
[2]	DIGANGI B A, GRAY L K, LEVY J K, et al. Detection of protective antibody titers against feline panleukopenia virus, feline herpesvirus-1, and feline calicivirus in shelter cats using a point-of-care ELISA[J]. J Feline Med Surg, 2011, 13(12):912-918.
[3]	DALL'ARA P, LABRIOLA C, SALA E, et al. Prevalence of serum antibody titres against feline panleukopenia, herpesvirus and calicivirus infections in stray cats of Milan, Italy[J]. Prev Vet Med, 2019, 167:32-38.
[4]	BERGMANN M, SPECK S, RIEGER A, et al. Antibody response to feline herpesvirus-1 vaccination in healthy adult cats[J]. J Feline Med Surg, 2020, 22(4):329-338.
[5]	HELPS C R, LAIT P, DAMHUIS A, et al. Factors associated with upper respiratory tract disease caused by feline herpesvirus, feline calicivirus, Chlamydophila felis and Bordetella bronchiseptica in cats:experience from 218 European catteries[J]. Vet Rec, 2005, 156(21):669-673.
[6]	FERNANDEZ M, MANZANILLA E G, LLORET A, et al. Prevalence of feline herpesvirus-1, feline calicivirus, Chlamydophila felis and Mycoplasma felis DNA and associated risk factors in cats in Spain with upper respiratory tract disease, conjunctivitis and/or gingivostomatitis[J]. J Feline Med Surg, 2017, 19(4):461-469.
[7]	LIU C H, LIU Y X, QIAN P, et al. Molecular and serological investigation of cat viral infectious diseases in China from 2016 to 2019[J]. Transbound Emerg Dis, 2020, 67(6):2329-2335.
[8]	SCHULZ C, HARTMANN K, MUELLER R S, et al. Sampling sites for detection of feline herpesvirus-1, feline calicivirus and Chlamydia felis in cats with feline upper respiratory tract disease[J]. J Feline Med Surg, 2015, 17(12):1012-1019.
[9]	HUSSEIN I T M, FIELD H J. Development of a quantitative real-time TaqMan PCR assay for testing the susceptibility of feline herpesvirus-1 to antiviral compounds[J]. J Virol Methods, 2008, 152(1-2):85-90.
[10]	MAZZEI M, VASCELLARI M, ZANARDELLO C, et al. Quantitative real time polymerase chain reaction (qRT-PCR) and RNAscope in situ hybridization (RNA-ISH) as effective tools to diagnose feline herpesvirus-1-associated dermatitis[J]. Vet Dermatol, 2019, 30(6):491-e147.
[11]	CHEN J, MA E B, HARRINGTON L, et al. CRISPR-Cas12a target binding unleashes indiscriminate single-stranded DNase activity[J]. Science, 2018, 360(6387):436-439.
[12]	BROUGHTON J P, DENG X D, YU G X, et al. CRISPR-Cas12-based detection of SARS-CoV-2[J]. Nat Biotechnol, 2020, 38(7):870-874.
[13]	HENZEL A, BRUM M C S, LAUTERT C, et al. Isolation and identification of feline calicivirus and feline herpesvirus in southern Brazil[J]. Braz J Microbiol, 2012, 43(2):560-568.
[14]	TOWNSEND W M, JACOBI S, TAI S H, et al. Ocular and neural distribution of feline herpesvirus-1 during active and latent experimental infection in cats[J]. BMC Vet Res, 2013, 9:185.
[15]	LIU M Z, HAN X H, YAO L Q, et al. Development and application of a simple recombinase polymerase amplification assay for rapid point-of-care detection of feline herpesvirus type 1[J]. Arch Virol, 2019, 164(1):195-200.
[16]	WANG J C, LIU L B, WANG J F, et al. Recombinase polymerase amplification assay-a simple, fast and cost-effective alternative to real time PCR for specific detection of Feline herpesvirus-1[J]. PLoS One, 2017, 12(1):e0166903.
[17]	TAN Y X, DONG G Y, XU H F, et al. Development of a cross-priming isothermal amplification assay based on the glycoprotein B gene for instant and rapid detection of feline herpesvirus type 1[J]. Arch Virol, 2020, 165(3):743-747.

RPA-Cas12a-LFD Based Nucleic Acid Detection for Feline Herpesvirus-1 and Preliminary Application

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References 17

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	WANG Jingyu, PAN Yangyang, XU Gengquan, ZHANG Rui, ZHANG Wenlan, WANG Xiaoshan, WU Rentaodi, ZHAO Rigetu, CUI Yan, YU Sijiu. Preparation and Preliminary Application of Yak (Bos Grunniens) Fas-associated Factor 1 Polyclonal Antibody [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(8): 3369-3382.
[2]	TIAN Yanhong, YU Jiangxu, JIAO Yuzhou, GAO Dongyang, CAI Xuwang. Research Progress on Structural Modification and Its Effects of Salmonella Lipopolysaccharides [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(4): 1392-1402.
[3]	XU Xinyan, ZHENG Yating, LIU Di, MA Bo, LIU Jiasen, QU Liandong. Isolation and Identification of a Feline Herpesvirus-1 Strain [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(4): 1713-1720.
[4]	SUN Dongxiao, ZHANG Shengli, ZHANG Qin, LI Jiao, ZHANG Guixiang, LIU Chousheng, ZHENG Weijie. Application Progress on Genomic Selection Technology for Dairy Cattle in China [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(10): 4028-4039.
[5]	SHAO Changxuan, DANG Ankai, ZHAN Zhaohan, LAI Zhenheng, ZHU Yongjie, SHAN Anshan. Development and Application Strategy of Beta Sheet Antimicrobial Peptides [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(8): 2490-2501.
[6]	JIANG Lingli, GAO Youling, HU Xingjuan, ZHANG Xian, YANG Yunkai, WEI Hua, FANG Weihuan. Development and Primary Application of Colloidal Gold-based Test Strip for Rapid Detection of Listeria monocytogenes LLO [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(6): 1841-1848.
[7]	DING Jun, FU Zilin, HE Junhao, DUAN Xiaowei, MA Lu, BU Dengpan. Research Progress of Milk-derived Exosomes [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(4): 1019-1029.
[8]	OUYANG Qingyuan, HU Shenqiang, WANG Jiwen. Current Genomics Research and Application of Important Traits in Poultry [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(3): 663-679.
[9]	ZOU Huiying, LI Junliang, ZHU Huabin. Progress on Research and Application of Prime Editing System [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(11): 3721-3730.
[10]	LI Yaqi, WANG Xianzhong, ZHANG Jiaojiao. Research Progress of Cell Proliferation and Apoptosis Regulated by Non-thermal Plasma and Its Application in Animal Husbandry [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(1): 1-10.
[11]	LI Zhanhong, ZHU Pei, SONG Ziang, LI Zhuoran, YANG Zhenxing, LI Huachun, YANG Heng, LIAO Defang. Establishment and Application of a Group Specific Reverse Transcriptase Loop-mediated Isothermal Amplification Method of Bluetongue Virus [J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(8): 2244-2253.
[12]	SU Kai, FAN Tingli, SONG Qinye. Ferritin Nanocarrier and Its Application Potential in the Field of Biomedicine [J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(4): 909-919.
[13]	XU Lu, ZHANG Qianyi, XIA Yingju, WANG Zhen, LI Cui, ZOU Xingqi, WANG Qin, ZHAO Qizu. Result Analysis of National Proficiency Testing Program for the Detection of Classical Swine Fever Virus Nucleic Acid [J]. Acta Veterinaria et Zootechnica Sinica, 2020, 51(8): 1949-1955.
[14]	LI Shuo, ZOU Tiande, WANG Zirui, YOU Jinming. The Metabolic Mechanism and Nutritional Physiological Function of α-ketoglutarate and Its Application in Livestock Production [J]. Acta Veterinaria et Zootechnica Sinica, 2020, 51(5): 923-932.
[15]	LIU Siyuan, LU Dan, TANG Zhonglin. Research Progress on CRISPR/Cas9 and Its Application in Pigs Genome Editing [J]. Acta Veterinaria et Zootechnica Sinica, 2020, 51(3): 409-416.