中国癌症防治杂志
中國癌癥防治雜誌
중국암증방치잡지
CHINESE JOURNAL OF ONCOLOGY PREVENTION AND TREATMENT
2013年
2期
117-121
,共5页
黄惠妮%吴华%沈菁%杨小丽%何敏
黃惠妮%吳華%瀋菁%楊小麗%何敏
황혜니%오화%침정%양소려%하민
液相芯片%肝癌标志物%AFP%方法
液相芯片%肝癌標誌物%AFP%方法
액상심편%간암표지물%AFP%방법
Liquid chip%Tumor marker%AFP%Method
目的建立液相芯片技术检测血清中AFP的反应体系，并对该法进行评价。方法采用双抗体夹心液相芯片技术，分别以不同偶联量的抗-AFP单抗与荧光微球偶联，以最适偶联量的荧光微球与不同浓度的生物素-羊抗鼠IgG反应，确定检测方法的线性范围、最低检测限、精密度等指标；应用液相芯片技术检测134例肝癌患者和47例健康对照者血清中AFP的浓度，并将该法与电化学发光免疫分析法（ECLIA）进行相关性分析。结果抗-AFP单抗的偶联最适量为8滋g，生物素-羊抗鼠IgG的最适浓度为4滋g/ml，检测AFP的线性范围为1.31~168 ng/ml，最低检测限为0.424 ng/ml，批内精密度为4.33%~7.66%，批间精密度为9.82%~13.85%。用液相芯片技术定量检测血清中AFP的浓度，各组差异有统计学意义（P<0.05）。液相芯片技术检测血清中AFP的灵敏度为85.82%，特异度为95.74%，该法与ECLIA法检测的相关系数r=0.845（P<0.05）。结论液相芯片技术检测血清中AFP与ECLIA法检测AFP有较好的相关性，并具有高通量、线性范围宽、灵敏度高、重复性好、节省样品和时间等优点，该法的建立将为进一步开发AFP联合检测多种肝癌标志物奠定基础。
目的建立液相芯片技術檢測血清中AFP的反應體繫，併對該法進行評價。方法採用雙抗體夾心液相芯片技術，分彆以不同偶聯量的抗-AFP單抗與熒光微毬偶聯，以最適偶聯量的熒光微毬與不同濃度的生物素-羊抗鼠IgG反應，確定檢測方法的線性範圍、最低檢測限、精密度等指標；應用液相芯片技術檢測134例肝癌患者和47例健康對照者血清中AFP的濃度，併將該法與電化學髮光免疫分析法（ECLIA）進行相關性分析。結果抗-AFP單抗的偶聯最適量為8滋g，生物素-羊抗鼠IgG的最適濃度為4滋g/ml，檢測AFP的線性範圍為1.31~168 ng/ml，最低檢測限為0.424 ng/ml，批內精密度為4.33%~7.66%，批間精密度為9.82%~13.85%。用液相芯片技術定量檢測血清中AFP的濃度，各組差異有統計學意義（P<0.05）。液相芯片技術檢測血清中AFP的靈敏度為85.82%，特異度為95.74%，該法與ECLIA法檢測的相關繫數r=0.845（P<0.05）。結論液相芯片技術檢測血清中AFP與ECLIA法檢測AFP有較好的相關性，併具有高通量、線性範圍寬、靈敏度高、重複性好、節省樣品和時間等優點，該法的建立將為進一步開髮AFP聯閤檢測多種肝癌標誌物奠定基礎。
목적건립액상심편기술검측혈청중AFP적반응체계，병대해법진행평개。방법채용쌍항체협심액상심편기술，분별이불동우련량적항-AFP단항여형광미구우련，이최괄우련량적형광미구여불동농도적생물소-양항서IgG반응，학정검측방법적선성범위、최저검측한、정밀도등지표；응용액상심편기술검측134례간암환자화47례건강대조자혈청중AFP적농도，병장해법여전화학발광면역분석법（ECLIA）진행상관성분석。결과항-AFP단항적우련최괄량위8자g，생물소-양항서IgG적최괄농도위4자g/ml，검측AFP적선성범위위1.31~168 ng/ml，최저검측한위0.424 ng/ml，비내정밀도위4.33%~7.66%，비간정밀도위9.82%~13.85%。용액상심편기술정량검측혈청중AFP적농도，각조차이유통계학의의（P<0.05）。액상심편기술검측혈청중AFP적령민도위85.82%，특이도위95.74%，해법여ECLIA법검측적상관계수r=0.845（P<0.05）。결론액상심편기술검측혈청중AFP여ECLIA법검측AFP유교호적상관성，병구유고통량、선성범위관、령민도고、중복성호、절성양품화시간등우점，해법적건립장위진일보개발AFP연합검측다충간암표지물전정기출。
Objective To develop liquid chip-based method for assaying alpha fetoprotein (AFP)in human serum and to evaluate method performance. Methods Different amounts of anti-AFP monoclonal antibody were coupled to fluorescent beads using liquid chip technology in a sandwich immunoassay format. Fluorescent microspheres prepared with the optimal amount of anti-AFP antibody were reacted with different concentrations of biotin-goat anti-mouse IgG,and several performance indicators were measured,including linear range,detection limit,and accuracy.The method was used to measure serum AFP concentration in 134 patients with hepatocellular carcino-ma(HCC)and 47 healthy controls,and the results were compared with those obtained by electrochemoluminescence immunoassay(ECLIA). Results Optimal coupling was obtained with 8μg of anti-AFP monoclonal antibody and 4μg/ml of biotin-goat anti-mouse IgG.The linear response range for the method to detect serum AFP was 1.31~168 ng/ml and the detection limit was 0.424 ng/ml. Intra- and inter-assay coefficients of variation ranged,respectively,from 4.33% to 7.66% and from 9.82% to 13.85%.The method detected significantly diffe-rent serum AFP levels between patients with HCC and healthy controls(P<0.05).Method sensitivity was 85.82% and specificity was 95.74%.Measurements obtained by the liquid chip method showed a correlation coefficient(r)of 0.845(P<0.05)with measurements obtained by ECLIA. Conclusion The liquid chip and ECLIA methods showed good correlation in detecting AFP.Since the liquid chip method offers a wider detection range and greater sensitivity than ECLIA,in addition to showing good repeatability,it may allow faster detection with fewer samples.This approach may also prove useful for detecting AFP together with other multiplexing markers in HCC.
