背景荧光猝灭免疫层析测定牛奶中林可霉素

目的：采用背景荧光猝灭-免疫层析法现场快速检测牛奶中林可霉素。方法：采用安装有Simple 1软件系统的背景荧光猝灭免疫分析仪，包括检测卡体系和背景荧光猝灭免疫分析仪，检测卡是其核心部件。基于背景荧光猝灭免疫层析法，建立牛奶中林可霉素的检测方法，制备了可用于快速检测的检测卡，同时进行方法学验证和样品测定。结果：确定检测卡最佳加样量为100 μL，最佳反应时间为10 min；检测卡的灵敏度为0.088 ng·mL^-1，线性范围曲线方程Y=-5.000×10^-6X³+0.000 4X²+0.001 6X+0.489 2，林可霉素在0.0~60 ng·mL^-1之间呈良好的相关性（r=0.992 6）。检测卡批间差RSD均小于2%；长期重复性试验RSD均小于1%。采用本方法实测牛奶样本，与国标方法检测结果比较，t检验结果P>0.05，说明采用背景荧光猝灭-免疫层析法和国标法2种方法比较没有显著性差异。结论：本法研制的检测卡专属性强，灵敏度高，在现场10 min内即可快速定量测定牛奶中林可霉素的含量，可应用于生鲜牛奶的质量控制及现场检测。

Objective: To establish a method of background fluorescence quenching immune chromatography (bFQIC) to determine lincomycin in milk. Methods: The instrument system of background fluorescence quenching immunochromatographic assay (bFQICA) used in this study included the test card system and the background fluorescence quenching immunoassay analyzer (Simple 1),The detection card was the core component.The detection method of lincomycin in milk was established based on background fluorescence quenching and immunochromatography,and a test card was developed for rapid detection,methodology validation and sample determination were performed simultaneously. Results: The best amount of sample quantity and react timing were 100 μL and 10 min. The sensitivity of the card was 0.088 ng·mL^-1,the linear equation was Y=-5.000× 10^-6X³+0.000 4X²+0.001 6X+0.489 2 (r=0.992 6),and the linear concentration range was 0.0-60 ng·mL^-1,the RSD was lower than 2% and the RSDs of long-term repetitive experiment were lower than 1%. t test of the results obtained from the proposed method and the international test method gave a significance of P>0.05,which illustrated that there was no difference between the methods. Conclusion: The test card developed in this study is specific and sensitive,which can be used for the rapid quantitative determination of lincomycin content in fresh milk within 10 minutes in the field,and can be used for the quality control and field detection of fresh milk.

[1] 张小辉,职爱民,侯玉泽,等.林可霉素人工抗原的合成与鉴定[J].西北农业学报,2012,21(8):1 ZHANG XH,ZHI AM,HOU YZ,et al.Synthesis and identification of lincomycin artifical antigen[J].Acta Agric Boreali-occident Sin,2012,21(8):1
[2] 叶兆伟,吴海港,刘锦妮,等.林可霉素制剂中林可霉素含量测定方法研究进展[J].信阳农林学院学报,2016,26(2):102 YE ZW,WU HG,LIU JN,et al.Research on the progress of lincomycin concet determination method[J].J Xinyang Agric Forestry Univ,2016,6(2):102
[3] 刘友饶,姜园园.林可霉素对开放性骨折患者预防感染的作用及其对血清学各指标的影响[J].抗感染药学,2017,14(1):194 LIU YR,JIANG YY.The effect of lincomycin on preventing infection in patients with open fracture and its effect on serological indexes[J].J Anti-Infec Pharm,2017,14(1):194
[4] 聂雯莹,罗晓琴,杜美红,等.林可霉素磁免疫化学发光检测方法的建立[J].山东畜牧兽医,2015,11(36):6 NIE WY,LUO XQ,DU MH,et al.Establishment of magnetoimmunochemiluminescence assay for lincomycin[J].Shandong J Anim Sci Veter Medi,2015,11(36):6
[5] HE J,WU N,LUO P,et al.Development of a heterologous enzyme-linked immunosorbent assay for the detection ofclindamycin and lincomycin residues in edible animal tissues[J].Meat Sci,2017,125:137
[6] CAO S,SONG S,LIU L,et a1.Comparison of anenzyme-linkedimmunosorbent assay with an immunoehromatographic assay for detection of lincomycinin milk and honey[J].Immunol Invest,2015,44(5):438
[7] LI SH,LIU CH,YIN GH,et al.Aptamer-molecularly imprinted sensor base on electrogenerated chemiluminescence energy transfer for detection of lincomycin[J].Biosens Bioelectron,2017, 91:687
[8] YANG C,ZHANG Z,SHI Z.A novel chemiluminescence reaction system for the determination of lincomycin with diperiodatonicke-late(IV)[J].Microchim Acta,2010,168(3):293
[9] 李维超,佟若菲,张囡,等.HPLC法测定复方盐酸林可霉素滴鼻剂中盐酸林可霉素和利巴韦林[J].现代药物与临床,2015,(1):33 LI WC,TONG RF,ZANG N,etal.Determination of lincomycin hydrochloride and ribavirin in compound lincomycin hydrochloride nasal drops by HPLC[J].Drugs Clin,2015,30(1):33
[10] SOLLIEC M,MASSE D,SAUVE S,et al.Analysis of trimethoprim,lincomycin,sulfadoxin and tylosin in swine manure using laser diode thermal desorption-atmospheric pressure chemical ionization-tandem mass spectrometry[J].Talanta,2014,128:23
[11] TAO Y,CHEN D,YU G,et al.Simultaneous determination of lincomycin and spectinomycin residues in animal tissues by gas chromatography-nitrogen phosphorus detection and gasehmmatography-mass spectrometrywith accelerated solvent extraction[J].Food Add Contamin:Part A,2001,28(2):145
[12] 赵福民,熊雅婷,钱博,等.乳品中多种抗生素残留胶体金检测方法研究[J].食品工业,2018,39(9):135 ZHAO FM,XIONG YT,QIAN B,et al.Simultaneous detection of multiple antibiotic residues in dairy products based on colloidal gold immunochromatography[J].Food Ind,2018,39(9):135
[13] 俞漪,胡雪莲,巢强国,等.液体乳中抗生素残留量快速检测方法探究[J].食品研究与开发,2010,31(8):133 YU Y,HU XL,CHAO QG,et al.Emulsified rapid detection of antibiotic residues method of inquiry[J].Food Res Devel,2010,31(8):133
[14] WU XX,TIAN XF,XU LH,et al.Determination of aflatoxin M1 and chloramphenicol in milk based on background fluorescence quenching immunochromatographic assay[J].Biomed Res Int,2017(3):45