中华检验医学杂志
中華檢驗醫學雜誌
중화검험의학잡지
CHINESE JOURNAL OF LABORATORY MEDICINE
2009年
11期
1262-1265
,共4页
谢靖婧%杨桂林%刘映霞%刘威龙%陈心春%朱秀云%徐六妹%周亚红%王火生%周伯平
謝靖婧%楊桂林%劉映霞%劉威龍%陳心春%硃秀雲%徐六妹%週亞紅%王火生%週伯平
사정청%양계림%류영하%류위룡%진심춘%주수운%서륙매%주아홍%왕화생%주백평
手足口病%肠道病毒A型%人%酶联免疫吸附测定%试剂盒%诊断
手足口病%腸道病毒A型%人%酶聯免疫吸附測定%試劑盒%診斷
수족구병%장도병독A형%인%매련면역흡부측정%시제합%진단
Hand%foot and mouth disease%Enterovims A%human%Enzyme-linkedimmunosorbent assay: Reagent kits%Diagnostic
目的 研制早期快速检测抗肠道病毒71型(EV71)抗体的ELISA血清学诊断性试剂盒,评价其临床应用价值.方法 将表达纯化的EV71重组蛋白VP1作为包被抗原,建立EV71型手足口病间接ELISA的血清学检测方法 .通过与逆转录(RT)PCR方法 、EV71病毒分离试验和微量血清中和试验比较,评价抗.EV71 IgM和抗-EV71 IgG血清学诊断方法 在EV71型手足口病的诊断价值.结果 与RT-PCR比较,抗-EV71 IgM敏感度、特异度、阳性预测值和阴性预测值分别为83%、85%、81%和87%;抗.EV71 IgG敏感度、特异度、阳性预测值和阴性预测值分别为72%、74%、68%和77%.与EV71病毒分离方法 比较,抗-EV71 IgM敏感度、特异度分别为85%和97%;抗-EV71 IgG敏感度、特异度分别为75%和77%.通过直线相关分析发现,抗-EV71 IgG抗体滴度和中和抗体滴度呈显著正相关(r=0.72,P＜0.05).EV71型手足口病患儿恢复期血清抗IgG滴度较急性期升高(P＜0.01),但抗IgM滴度差异无统计学意义(P＞0.05).结论 利用VP1重组蛋白作为包被抗原,成功开发ELISA检测人血清抗-EV71 IgM和抗-EV71 IgG抗体的诊断试剂盒.
目的 研製早期快速檢測抗腸道病毒71型(EV71)抗體的ELISA血清學診斷性試劑盒,評價其臨床應用價值.方法 將錶達純化的EV71重組蛋白VP1作為包被抗原,建立EV71型手足口病間接ELISA的血清學檢測方法 .通過與逆轉錄(RT)PCR方法 、EV71病毒分離試驗和微量血清中和試驗比較,評價抗.EV71 IgM和抗-EV71 IgG血清學診斷方法 在EV71型手足口病的診斷價值.結果 與RT-PCR比較,抗-EV71 IgM敏感度、特異度、暘性預測值和陰性預測值分彆為83%、85%、81%和87%;抗.EV71 IgG敏感度、特異度、暘性預測值和陰性預測值分彆為72%、74%、68%和77%.與EV71病毒分離方法 比較,抗-EV71 IgM敏感度、特異度分彆為85%和97%;抗-EV71 IgG敏感度、特異度分彆為75%和77%.通過直線相關分析髮現,抗-EV71 IgG抗體滴度和中和抗體滴度呈顯著正相關(r=0.72,P＜0.05).EV71型手足口病患兒恢複期血清抗IgG滴度較急性期升高(P＜0.01),但抗IgM滴度差異無統計學意義(P＞0.05).結論 利用VP1重組蛋白作為包被抗原,成功開髮ELISA檢測人血清抗-EV71 IgM和抗-EV71 IgG抗體的診斷試劑盒.
목적 연제조기쾌속검측항장도병독71형(EV71)항체적ELISA혈청학진단성시제합,평개기림상응용개치.방법 장표체순화적EV71중조단백VP1작위포피항원,건립EV71형수족구병간접ELISA적혈청학검측방법 .통과여역전록(RT)PCR방법 、EV71병독분리시험화미량혈청중화시험비교,평개항.EV71 IgM화항-EV71 IgG혈청학진단방법 재EV71형수족구병적진단개치.결과 여RT-PCR비교,항-EV71 IgM민감도、특이도、양성예측치화음성예측치분별위83%、85%、81%화87%;항.EV71 IgG민감도、특이도、양성예측치화음성예측치분별위72%、74%、68%화77%.여EV71병독분리방법 비교,항-EV71 IgM민감도、특이도분별위85%화97%;항-EV71 IgG민감도、특이도분별위75%화77%.통과직선상관분석발현,항-EV71 IgG항체적도화중화항체적도정현저정상관(r=0.72,P＜0.05).EV71형수족구병환인회복기혈청항IgG적도교급성기승고(P＜0.01),단항IgM적도차이무통계학의의(P＞0.05).결론 이용VP1중조단백작위포피항원,성공개발ELISA검측인혈청항-EV71 IgM화항-EV71 IgG항체적진단시제합.
Objective To develop an ELISA(Enzyme-Linked Immunosorbent Assay)diagnostic kit for early rapid detection of sarum anti-EV71 antibody and evaluate its clinical application value.Methods Recombinant protein VP1 of EV71 were prepared and purified as an immobilized antigen for establishment of an indirect ELISA for detection of serum anti-EV71 IgM and anti-EV71 IgG.Compared with RT-PCR.isolation of EV71 and micro-neutralizing assay.the clinical application value of anti-EV71 IgM and anti-EV71 ISG in the diagnosis of EV71 disease was evaluated.Results In comparison with RT-PCR.the sensitivity,specificity,positive predictive value and negative predictive value of anti-EV71 IgM antibody were 83%,85%,81%and 87%,respectively.The sensitivity,specificity,positive predictive value and negative predictive value of anti-EV71 IgG antibody were 72%,74%,68%and 77%.respectively.Compared with viral isolation assay.the sensitivity and specificity of anti-EV71 IgM antibody were 85%and 97%,respectively.The sensitivity and specificity of anti-EV71 IsG antibody were 75%and 77%,respectively.In addition.the titers of anti-EV71 IgG antibody were significantly correlated with the titers of neutralizing antibody to EV71 by linear regression analysis(r=0.72,P＜0.05).Finally,the serum titers of anti-IgG from patients with EV71 associated hand food and mouth disease at convalescent stage exhibited significantly higher than that of the same patients at acute stage(P＜0.01),but the titers of anti-IgM had no significant difference(P＞0.05).Conclusions With VP1 recombinant protein used as an immobilized antigen,an indirect ELISA diagnostic kit was successfully develooed for detection of serum anti-human EV71 IgM and anti-human EV71 IgG antibodies.
