犬C-反应蛋白荧光定量免疫层析检测方法的建立与应用

doi:10.16431/j.cnki.1671-7236.2019.02.034

Abstract

Abstract:

The study focused on the establishment of fluorescent microsphere immunochromatography quantitative method of canine C-reactive protein (C-CRP) to a develop test strips of simple operation and high sensitivity for rapid quantitative detection of C-CRP.A fluorescent sandwich immunoassay via double antibody was set up,anti-C-CRP monoclonal antibody and goat anti-chicken IgY labeled with carboxy fluorescent microspheres were used as labeled antibody,the other anti-C-CRP monoclonal antibody and chicken IgY were used as capture antibody for detection lines and control line.The results showed that the detection range of the lateral flow assay was 0.5 to 250 mg/L,the detection limit was 0.5 mg/L,and the CV in intra-assay and inter-assay were 0.68% to 6.94% and 0.89% to 8.79%,respectively.The average recovery was 98.15% to 101.19%,and there was no cross-react with 9 interfering substances.The stressed condition stability test showed that the test strip could be stored for 24 months at room temperature.For clinical sample test,the test strips had a good correlation with the control reagent (R²=0.995).In conclusion,the prepared fluorescent immunochromatographic strips could be applied to clinical test of C-CRP concisely and conveniently.

Key words: canine C-reactive protein(C-CRP); fluorescence immunochromatography; quantitative detection

CLC Number:

S858.292

KANG Ye, WANG Yu, HU Shukun, WU Peidian, LI Wenmei, HE Xiaowei. Establishment and Application of Fluorescence Quantitive Immunochromatographic Assay for Canine C-reactive Protein[J]. , 2019, 46(2): 621-627.

References

[1] 王心竹.浅谈我国宠物行业的发展现状及趋势[J].现代畜牧兽医,2018,2:57-59. WANG X Z.Current situation and development trend of pet industry in China[J].Modern Journal of Animal Husbandry and Veterinary Medicine,2018,2:57-59.(in Chinese)
[2] 王怡君.犬C-反应蛋白免疫透射比浊定量检测方法的建立[D].长春:吉林农业大学,2013. WANG Y J.Establish of immune turbidmetric quantitative detection method for canine C-reactive protein[D].Changchun:Jilin Agricultural University,2013.(in Chinese)
[3] 张海霞,夏兆飞.犬C-反应蛋白的研究进展[J].中国兽医杂志,2010,46(12):70-71. ZHANG H X,XIA Z F.Research progress on canine C-reactive protein[J].Chinese Journal of Veterinary Medicine,2010,46(12):70-71.(in Chinese)
[4] SCHMIDT E M D S,ECKERSALL P D.Acute phase proteins as markers of infectious diseases in small animals[J].Acta Veterinaria,2015,65(2):149-161.
[5] 刘洋.犬C-反应蛋白在疾病诊疗中的应用[D].呼和浩特:内蒙古农业大学,2015. LIU Y.Application of canine C-reactive protein in diagnosis[D].Hohhot:Inner Mongolia Agricultural University,2015.(in Chinese)
[6] ONISHI T,INOKUMA H,OHNO K,et al.C-reactive protein concentrations in normal and diseased dogs-measured by laser nephelometric immunoassay[J].Journal of the Japan Veterinary Medical Association,2000,53(9):595-601.
[7] 陈艳云,夏兆飞.犬C反应蛋白研究进展[J].中国兽医杂志,2016,52(2):61-63. CHEN Y Y,XIA Z F.Research progress on canine C-reactive protein[J].Chinese Journal of Veterinary Medicine,2016,52(2):61-63.(in Chinese)
[8] ECKERSALL P D.Acute phase protein in dogs and cats:Current knowledge and future perspectives[J].Veterinary Clinical Pathology,2005,34(2):85-99.
[9] ECKERSALL P D,BELL R.Acute phase proteins:Biomarkers of infection and inflammation in veterinary medicine[J].Veterinary Journal,2010,185(1):23-27.
[10] KJELGAARD-HANSEN M,JENSEN A L,KRISTENSEN A T.Internal quality control of a turbid metric immunoassay for canine serum C-reactive protein based on pooled patient samples[J].Veterinary Clinical Pathology,2010,33(3):139-144.
[11] 吴伟兵,王明亮,景宜,等.单分散荧光微球的制备及其光学性能研究[J].南京林业大学学报(自然科学版),2010,34(3):15-19. WU W B,WANG M L,JING Y,et al.Preparation of monodisperse fluorescent microspheres and their optical properties[J].Journal of Nanjing Forestry University (Natural Science Edition),2010,34(3):15-19.(in Chinese)
[12] 刘道峰,邓省亮,赖卫华,等.莱克多巴胺荧光微球免疫层析检测方法的建立[J].食品与机械,2012,28(1):73-77. LIU D F,DENG S L,LAI W H,et al.Development of lateral flow assay of ractopamine based on fluorescent microspheres[J].Food & Machinery,2012,28(1):73-77.(in Chinese)
[13] 张赛,洪裕好,李凯,等.尿微量白蛋白荧光免疫层析定量检测试剂的研制及性能评价[J].生物技术通报,2017,33(3):199-204. ZHANG S,HONG Y H,LI K,et al.Establishment and performance evaluation of a quantitative detection method for microalbuminuria based on fluorescence immunochromatography[J].Biotechnology Bulletin,2017,33(3):199-204.(in Chinese)
[14] 王凡,白羽,于咏兰.C-反应蛋白检测在犬多种疾病中的应用[J].动物医学进展,2018,39(6):108-112. WANG F,BAI Y,YU Y L.Detection of C-reaction protein in canine with some diseases[J].Progress in Veterinary Medicine,2018,39(6):108-112.(in Chinese)
[15] CHRISTENSEN M B,ERIKSEN T,KJELGAARDHANSEN M.C-reactive protein:Quantitative marker of surgical trauma and post-surgical complications in dogs:A systematic review[J].Acta Veterinaria Scandinavica,2015,57(1):71.
[16] KOCATURK M,TVARIJONAVICIUTE A,MARTINEZSUBIELA S,et al.Inflammatory and oxidative biomarkers of disease severity in dogs with parvoviral enteritis[J].Journal of Small Animal Practice,2015,56(2):119-124.
[17] SATO T,OHNO K,TAMAMOTO T,et al.Assesment of severity and changes in C-reactive protein concentration and various biomarkers in dogs with pancreatitis[J].Journal of Veterinary Medical Science,2017,79(1):35-40.
[18] PARRA M D,TUOMOLA M,CABEZASHERRERA J,et al.Analytical and clinical validation of a time-resolved immunofluorometric assay (TR-IFMA) for canine C-reactive protein in serum[J].Veterinary Research Communications,2006,30(2):113-126.
[19] IBRAIMI F,EKBERG B,KRIZ D,et al.Preparation of a portable point-of-care in vitro diagnostic system,for quantification of canine C-reactive protein,based on a magnetic two-site immunoassay[J].Analytical & Bioanalytical Chemistry,2013,405(18):6001-6007.
[20] 万宇平,龙光宗,吴小胜,等.一种犬C反应蛋白荧光检测试纸条[P].2017. WAN Y P,LONG G Z,WU X S,et al.One kind of fluorescent dog C-reactive protein test strip[P].2017.(in Chinese)
[21] PINERO M,PATO R,SOLER L,et al.A new automated turbidimetric immunoassay for the measurement of canine C-reactive protein[J].Veterinary Clinical Pathology,2018,47(1):130-137.

Establishment and Application of Fluorescence Quantitive Immunochromatographic Assay for Canine C-reactive Protein

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 4

Recommended Articles

Metrics

Comments

[1]	WU Meng, WEI Zhijing, FAN Yiming, CUI Chenguang, LI Shuang, DONG Weiya. Development of a Time-resolved Fluorescence Immunochromatography Assay Kit for Detection of Bovine Serum Amyloid A [J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(4): 1191-1198.
[2]	XIA Wenlong, WU Zhi, GUO Changming, ZHU Shanyuan, YU Shupei, ZHANG Xinyu, XIA Xiaoli, SUN Huaichang. Establishment and Application of Sandwich ELISA for Quantitative Detection of Porcine Reproductive and Respiratory Syndrome Virus Soluble Receptors [J]. , 2018, 45(4): 881-887.
[3]	YANG Song;LIANG Cheng-zhu;GAO Hong-wei;CHEN Pu-yan. Detection of Equine Arteritis Virus by Real-time TaqMan Reverse Transcription-PCR Assay [J]. , 2011, 38(9): 115-118.
[4]	LI Meng;XU Hui-hui;GAO Jin-yu;LEI Lian-cheng. Development of an Optimized ELISA for the Quantitative Detection of Protein in the Milk [J]. , 2011, 38(4): 84-87.