中华检验医学杂志
中華檢驗醫學雜誌
중화검험의학잡지
CHINESE JOURNAL OF LABORATORY MEDICINE
2014年
7期
544-547
,共4页
棘球蚴病%酶联免疫吸附测定%试剂盒,诊断%敏感性与特异性
棘毬蚴病%酶聯免疫吸附測定%試劑盒,診斷%敏感性與特異性
극구유병%매련면역흡부측정%시제합,진단%민감성여특이성
Echinococcosis%Enzyme-linked immunosorbent assay%Reagent kits,diagnostic%Sensitivity and specificity
目的 采用正交实验筛选包虫病ELISA试剂盒最佳条件及试剂盒制备,检测其检验效能.方法 以检测样品吸光度为评价指标,对试剂盒抗原包被浓度(A)、样品稀释倍数(B)、酶标抗原稀释倍数(C)3个因素的4个水平进行L16(43)正交实验,筛选最佳条件,并由新疆医科大学第一附属医院标本库提供2011年3月至2013年6月收集的36份经金标准确认的泡状棘球蚴患者血清,56份经金标准确认的细粒棘球蚴患者血清,72份对照组血清(包括健康人、肝硬化、肝炎等).检测其敏感度、特异度,采用F检验进行统计学分析.结果 正交设计结果显示包虫病ELISA试剂盒的3个影响因素大小依次是:酶标抗原稀释倍数＞样品稀释倍数＞抗原包被浓度,最佳制备条件A1B2C4,即抗原包被浓度1 mg/L,样品稀释倍数1∶100,酶标抗原稀释倍数1∶160000,为最佳条件.通过对试剂盒3个因素4水平进行正交设计,比较各组合之间的吸光度,可见因素抗原包被浓度(A),F=1.181,P=0.393,因素样品稀释倍数(B),F=2.544,P =0.152,因素酶标抗原稀释倍数(C),F=2.544,P=0.039.本试剂盒对细粒棘球蚴病和泡状棘球蚴病的敏感度分别是83％(30/36)、91％ (51/56);特异度为91％(29/32)、83％ (33/40);漏诊率17％(6/36)、9％(5/56);误诊率9％(3/32)、18％ (7/40);阳性似然比8.86(83/9)、5.20(91/18);阴性似然比0.18(17/91)、0.11(9/83).结论 正交设计有良好的发展前景,能够有效简便的筛选出制备包虫病ELISA试剂盒的最优条件,提高其检验效能.
目的 採用正交實驗篩選包蟲病ELISA試劑盒最佳條件及試劑盒製備,檢測其檢驗效能.方法 以檢測樣品吸光度為評價指標,對試劑盒抗原包被濃度(A)、樣品稀釋倍數(B)、酶標抗原稀釋倍數(C)3箇因素的4箇水平進行L16(43)正交實驗,篩選最佳條件,併由新疆醫科大學第一附屬醫院標本庫提供2011年3月至2013年6月收集的36份經金標準確認的泡狀棘毬蚴患者血清,56份經金標準確認的細粒棘毬蚴患者血清,72份對照組血清(包括健康人、肝硬化、肝炎等).檢測其敏感度、特異度,採用F檢驗進行統計學分析.結果 正交設計結果顯示包蟲病ELISA試劑盒的3箇影響因素大小依次是:酶標抗原稀釋倍數＞樣品稀釋倍數＞抗原包被濃度,最佳製備條件A1B2C4,即抗原包被濃度1 mg/L,樣品稀釋倍數1∶100,酶標抗原稀釋倍數1∶160000,為最佳條件.通過對試劑盒3箇因素4水平進行正交設計,比較各組閤之間的吸光度,可見因素抗原包被濃度(A),F=1.181,P=0.393,因素樣品稀釋倍數(B),F=2.544,P =0.152,因素酶標抗原稀釋倍數(C),F=2.544,P=0.039.本試劑盒對細粒棘毬蚴病和泡狀棘毬蚴病的敏感度分彆是83％(30/36)、91％ (51/56);特異度為91％(29/32)、83％ (33/40);漏診率17％(6/36)、9％(5/56);誤診率9％(3/32)、18％ (7/40);暘性似然比8.86(83/9)、5.20(91/18);陰性似然比0.18(17/91)、0.11(9/83).結論 正交設計有良好的髮展前景,能夠有效簡便的篩選齣製備包蟲病ELISA試劑盒的最優條件,提高其檢驗效能.
목적 채용정교실험사선포충병ELISA시제합최가조건급시제합제비,검측기검험효능.방법 이검측양품흡광도위평개지표,대시제합항원포피농도(A)、양품희석배수(B)、매표항원희석배수(C)3개인소적4개수평진행L16(43)정교실험,사선최가조건,병유신강의과대학제일부속의원표본고제공2011년3월지2013년6월수집적36빈경금표준학인적포상극구유환자혈청,56빈경금표준학인적세립극구유환자혈청,72빈대조조혈청(포괄건강인、간경화、간염등).검측기민감도、특이도,채용F검험진행통계학분석.결과 정교설계결과현시포충병ELISA시제합적3개영향인소대소의차시:매표항원희석배수＞양품희석배수＞항원포피농도,최가제비조건A1B2C4,즉항원포피농도1 mg/L,양품희석배수1∶100,매표항원희석배수1∶160000,위최가조건.통과대시제합3개인소4수평진행정교설계,비교각조합지간적흡광도,가견인소항원포피농도(A),F=1.181,P=0.393,인소양품희석배수(B),F=2.544,P =0.152,인소매표항원희석배수(C),F=2.544,P=0.039.본시제합대세립극구유병화포상극구유병적민감도분별시83％(30/36)、91％ (51/56);특이도위91％(29/32)、83％ (33/40);루진솔17％(6/36)、9％(5/56);오진솔9％(3/32)、18％ (7/40);양성사연비8.86(83/9)、5.20(91/18);음성사연비0.18(17/91)、0.11(9/83).결론 정교설계유량호적발전전경,능구유효간편적사선출제비포충병ELISA시제합적최우조건,제고기검험효능.
Objective To select the optimum conditions of Echinococcosis ELISA kit and kit preparation by using orthogonal experiments,then to test their detection performance.Methods Taking the absorbance of the samples as the indicator,orthogonal experiments of three factors four levels were taken to optimize the conditions of hydatid disease detection ELISA kit.Through L16 (43)orthogonal experiments,the concentration of the antigen(A),the dilution multiple of the sample (B) and the dilution multiple of the enzyme labeled antigen (C) were studied.The specimen were provided by The First Affiliated Hospital of Xinjiang Medical University specimen library collected between March 2011 and June 2013,including 36 sera confirmed alveolar Echinococcus by the gold standard,56 sera confirmed Echinococcus granulosus disease by the gold standard and 72 sera as the control group(including healthy people,cirrhosis,hepatitis,etc).Detecting sensitivity and specificity were compared using F test for statistical analysis.Results Orthogonal design showed the size proportion of three factors of echinococcosis ELISA kit:antibody dilution ＞ sample dilution ＞ antigen coating concentration.The optimal preparation conditions were A1 B2 C4,that was,the concentration of antigen was 1 mg/L,sample dilution of 1∶ 100,dilution of secondary antibody of 1∶160 000.Through the 3-factor and 4-level orthogonal design using F-test analysis,the fact of the concentration of the kit antigen(A),F =1.181,P =0.393; the fact of the dilution multiple of the sample (B),F =2.544,P =0.152,the fact of enzyme labeled antigen dilution (C) F =2.544,P =0.039.The sensitivity of the kit to Echinococcus granulosus disease and alveolar Echinococcus were 83％ (30/36),and 91％ (51/56),respectively; the specificity was 91％ (29/32) and 83％ (33/40); the misdiagnosis rate of 17％ (6/36) and 9％ (5/56) ; the misdiagnosis rate 9％ (3/32) and 18％ (7/40) ; positive likelihood ratio 8.86％ (83/9) and 5.20％ (91/18) ; negative likelihood ratio of 0.18％ (17/91) and 0.11％ (9/83).Conclusion Orthogonal design is a good method that can find out optimal conditions of preparation for Echinococcosis ELISA kit and improve the test performance.
