| Cardiovascular disease(CVD)is the name for the group of disorders of heart and blood vessels,which includes coronary heart disease,cerebrovascular disease,peripheral vascular disease,cardiomyopathies,CVDs are the number one cause of death globally,more people die annually from CVDs than from any other cause.An estimated 17.5 million people died from CVDs in 2012,representing 31%of all global deaths,of these deaths,an estimated 7.4million were due to coronary heart disease and 6.7 million were due to stroke,and the trend of the mortality rate is increasing year by year.The early diagnosis of cardiovascular disease is of great significance for prevention and monitoring of disease,and the rapid diagnosis ofcardiovascular predicting factors is arousing more and moreattention of people.In recent years,a large number of studies have confirmed that homocysteine(Hcy)and high-sensitivity C-reactive protein(hs-CRP)are closely related to various cardiovascular diseases.Particularly,the concentration of Hcy and hs-CRP are considered as two independent risk factors of cardiovascular diseases.In this study we try to establish a lateral flow immunoassay test strip that can quantitive detect these two factors,the main research content aredescribed as below.(1)A rapid and easy to operate point-of-care testing strip for the detection of Hcy was developed,which was based on the competitive immunochromatography,Biotin-antibody was fixed on the conjugate pad to specifically capture S-adenosyl homocysteine(SAH)in the pretreated sample,where SAH was transformed from S-Adenosyl methionine(SAM)by homocysteine S-methyltransferase(HMT)catalysis using Hcy as substrate,the concentration of SAH was proportional to that of Hcy.The detection limit of 0.26?M Hcy was achieved with a good linear relationshipfrom 1.0 to 50.0?mol/L.Besides,the assay possess good reproducibility(CV<10%).Furthermore,other structural analogs SAM and cysteine(Cys)showed negative results,validating the excellent specificity of our method.(2)Alateral flow immunofluorescence test stripfor the detection of hs-CRP was developed based on double antibodies sandwich immunochromatographic assay.The labeled antibody Ab1 was ligated to the fluorescent microspheres to form a probe,which was fixed at the front end of the nitrocellulose membrane in an amount of 1?L/cm,the concentration of the antibody Ab2 on the detecting line was 1 mg/mL.The detection linear range of the test strip was 0.3-10 mg/L,and the detection limit was 0.107 mg/L.The test strip possessed good detection reproducibility(CV<10%).The detection accuracy of human serum samples was good compared with high performance liquid chromatography(HPLC,R~2>0.99)and had good stability after 10 days'storage at 37°C.(3)A dual-parameter test strip capable of early vascular inflammation rapid diagnostic was established.The test strip had good detection linearity and could detect Hcy well,the detected concentration of hs-CRP was lower than actual concentration because of the pretreatment process of Hcy.The reproducibility of this dual-index test strip of Hcy and hs-CRP was explored at low,medium and high concentration samples,the result showed that the test strip possessed good reproducibility(CV<10%)and detection stability. |
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