中华结核和呼吸杂志
中華結覈和呼吸雜誌
중화결핵화호흡잡지
Chinese Journal of Tuberculosis and Respiratory Diseases
2011年
5期
356-358
,共3页
范超明%李伟%樊德利%郑小银%丁建祖
範超明%李偉%樊德利%鄭小銀%丁建祖
범초명%리위%번덕리%정소은%정건조
分枝杆菌,结核%胶体金%双抗原夹心法%免疫层析法
分枝桿菌,結覈%膠體金%雙抗原夾心法%免疫層析法
분지간균,결핵%효체금%쌍항원협심법%면역층석법
Mycobacterium tuberculosis%Gold colloid%Double antigen eontmns filling the law%Immunochromatographic analysis
目的 建立一种简易、快速和准确的双抗原夹心胶体金免疫层析法,用于检测结核病患者血清中的抗MTB抗体.方法 采用胶体金标记相对分子质量分别为6000、16 000和38 000的重组MTB早期分泌抗原靶蛋白(ESAT),成为重组MTB融合蛋白ESAT-16-38,研制出双抗原胶体金免疫层析检测卡,对浙江省德清县疾病预防控制中心2007-2009年临床确诊的结核病患者血清163份(其中肺结核痰MTB阳性57份、痰MTB阴性64份和肺外结核42份)和对照血清573份(其中体检者血清224份、急性肺炎和支气管炎患者血清217份、肺吸虫患者血清132份)样本进行检测,并与MTB蛋向芯片抗体法的检测结果进行比较.检出率的比较采用χ2检验.结果 双抗原法和蛋白芯片法检测结核病患者血清的阳性符合率分别为73.0%(120/163)和72.4%(118/163),差异无统计学意义(χ2=0.062,P＞0.05);检测对照血清的阴性检出率分别为93.9%(538/573)和92.0%(527/573),差异无统计学意义(χ2=0.635,P＞0.05);未见与肺吸虫患者血清有交叉反应,双抗原法对痰MTB阳性血清的检出率高达87.7%(50/57).结论 重组ESAT-6和ESAT-16双抗原胶体金免疫层析检测卡具有较高的敏感度和特异度,与蛋白芯片法的检测结果相似,且方法简便、快速,有很好的重现性和稳定性.
目的 建立一種簡易、快速和準確的雙抗原夾心膠體金免疫層析法,用于檢測結覈病患者血清中的抗MTB抗體.方法 採用膠體金標記相對分子質量分彆為6000、16 000和38 000的重組MTB早期分泌抗原靶蛋白(ESAT),成為重組MTB融閤蛋白ESAT-16-38,研製齣雙抗原膠體金免疫層析檢測卡,對浙江省德清縣疾病預防控製中心2007-2009年臨床確診的結覈病患者血清163份(其中肺結覈痰MTB暘性57份、痰MTB陰性64份和肺外結覈42份)和對照血清573份(其中體檢者血清224份、急性肺炎和支氣管炎患者血清217份、肺吸蟲患者血清132份)樣本進行檢測,併與MTB蛋嚮芯片抗體法的檢測結果進行比較.檢齣率的比較採用χ2檢驗.結果 雙抗原法和蛋白芯片法檢測結覈病患者血清的暘性符閤率分彆為73.0%(120/163)和72.4%(118/163),差異無統計學意義(χ2=0.062,P＞0.05);檢測對照血清的陰性檢齣率分彆為93.9%(538/573)和92.0%(527/573),差異無統計學意義(χ2=0.635,P＞0.05);未見與肺吸蟲患者血清有交扠反應,雙抗原法對痰MTB暘性血清的檢齣率高達87.7%(50/57).結論 重組ESAT-6和ESAT-16雙抗原膠體金免疫層析檢測卡具有較高的敏感度和特異度,與蛋白芯片法的檢測結果相似,且方法簡便、快速,有很好的重現性和穩定性.
목적 건립일충간역、쾌속화준학적쌍항원협심효체금면역층석법,용우검측결핵병환자혈청중적항MTB항체.방법 채용효체금표기상대분자질량분별위6000、16 000화38 000적중조MTB조기분비항원파단백(ESAT),성위중조MTB융합단백ESAT-16-38,연제출쌍항원효체금면역층석검측잡,대절강성덕청현질병예방공제중심2007-2009년림상학진적결핵병환자혈청163빈(기중폐결핵담MTB양성57빈、담MTB음성64빈화폐외결핵42빈)화대조혈청573빈(기중체검자혈청224빈、급성폐염화지기관염환자혈청217빈、폐흡충환자혈청132빈)양본진행검측,병여MTB단향심편항체법적검측결과진행비교.검출솔적비교채용χ2검험.결과 쌍항원법화단백심편법검측결핵병환자혈청적양성부합솔분별위73.0%(120/163)화72.4%(118/163),차이무통계학의의(χ2=0.062,P＞0.05);검측대조혈청적음성검출솔분별위93.9%(538/573)화92.0%(527/573),차이무통계학의의(χ2=0.635,P＞0.05);미견여폐흡충환자혈청유교차반응,쌍항원법대담MTB양성혈청적검출솔고체87.7%(50/57).결론 중조ESAT-6화ESAT-16쌍항원효체금면역층석검측잡구유교고적민감도화특이도,여단백심편법적검측결과상사,차방법간편、쾌속,유흔호적중현성화은정성.
Objective To establish a simple,fast and accurate double antigen colloidal gold immunochromatographic technique for detecting Mycobacterium tuberculosis antibody from tuberculosis patients.Methods The fusion protein ESAT-6-16-38 was constructed by using gene cloning technique for the 6,16 and 38 kDa early secreted antigenic target from Mycobacterium tuberculosis.The ESAT-6-16-38 fusion protein was marked to colloidal gold to eatablish the double antigen colloidal gold immunochromatographie assay.Serum samples from 163 patients with tuberculosis,including 57 sputumpositive cases,64 sputum-negative cases,and 42 cases with extrapulmonary tuberculosis,were collected during 2007and 2009 from the Disease Prevention and Control Center of Deqing County.In addition,573 controls(224 healthy volunteers,217 patients with acute pneumonia and bronchitis,132 patients with paragonimiasis)were recruited for comparison.Mycobacterium tuberculosis specific antibodies were detected by using immunochromatographic and protein chip technique.Detection rate was compared with Chi-square test. Results Among the 163 tuberculosis patients,the positive rates of immunochmmatographic detection and protein chip were 73.0%(120/163)and 72.4%(118/163)respectively;the difference was not statistically significant(χ2=0.062,P＞0.05).Among the 573 controls,the negative rates of immunochromatographic detection and protein chip were 93.9%(538/573)and 92.0%(527/573)respectively;the difference was not statistically significant(χ2=0.635,P＞0.05).There was no cross reaction in the paragonimiasis patients.The positive rate of the immunoehromatographie assay was as high as 87.7%(50/57)in the sputmn-positive patients.Conclusions The double antigen immunoehromatographie technique is an easy to operate, rapid,highly sensitive, specific, and reproducible method for the detection of Mycobacterium tuberculosis antibodies.
