幽门螺杆菌粪便抗原纳米金侧流免疫层析快速检测板的研制及应用评价  

Evaluation of the Development and Application of Rapid Testing Plate with Gold Nanoparticles Lateral-flow Immunochromatographic for Detecting Helicobacter Pylori Stool Antigen

作　　者：陈达伟[1] 江晓肖[1] 戴玉柱[2] 陈少明[1] 王国政[2] 孙长贵[2] 成军[2] 

机构地区：[1]解放军第117医院消化科,杭州310013 [2]解放军第117医院临床实验中心,杭州310013

出　　处：《现代检验医学杂志》2013年第3期75-77,共3页Journal of Modern Laboratory Medicine

摘　　要：目的 研制幽门螺杆菌（HP）粪便抗原（HPSA）纳米金侧流免疫层析快速检测板.方法 利用枸橼酸钠作为还原剂制备纳米金,标记兔抗HP多价抗体,建立双抗体夹心法的纳米金侧流免疫层析快速检测板;采用该检测板和英国Atlas公司试纸条分别对HP阳性病例组（123例）、健康体检HP阳性对照I组（109例）、健康体检HP阴性对照II组（129例）进行双盲法HPSA检测,并以14C-尿素呼气试验（14C-UBT）为金标准,对结果进行评价.结果 制备的直径40 nm纳米金最大吸收波长（λmax）为525 nm,标记抗体的0.05 mol/L硼酸盐缓冲液最适pH为7.6,抗体标记最佳用量为18 μg/ml（最小标记量为15 μg/ml）;检测板与试纸条的检测结果在三组之间差异无统计学意义（χ^2=0～0.337,P均〈0.05）;以14C-UBT结果为金标准,检测板的灵敏度、特异度、符合率分别为98.3%,99.2%和98.6%,试纸条的灵敏度、特异度、符合率分别为97.4%,98.4%和97.8%.结论 以纳米金标记多抗的双抗体夹心法侧流免疫层析快速检测板,检测HPSA诊断HP感染,方法简便、价格便宜、重复性好、无创伤,具有很高的敏感度和特异度.Objective To develop rapid testing plate of gold nanoparticles lateral-flow immunoehromatographic for detecting helicobacter pylori stool antigen（HPSA）. Methods The rabbit anti-HP polyvalent antibody was labeled by gold nanoparticles, which was synthetized using three citric acid sodium as reducing agent. Rapid testing plate for detecting HPSA was de- veloed with gold nanoparticles lateral-flow immunochromatographic technique of double antibody sandwich. HPSA were double-blind detected by the rapid testing plate and testing strip of the British Atlas company in 123 patients with HP posi- tive（case group）, 109 cases of healthy with HP positive（control group I）, 129 cases of healthy with HP negative （control group II） respectively. And the results were evaluated with 14C urea breath test （14 C-UBT） as the gold standard. Results The maximum absorption wavelength （kmax） of gold nanoparticles with 40 nm diameter was 525 nm. The optimum pH val- ue of 0.05 mol/L borate buffer,the optimum and minimum amount of labeled antibody were 7.6,18 t^g/ml and 15 t^g/ml,re- speetively. The results of rapid testing plate and testing strip for detecting HPSA had no statistically significant differences among the case group,control group I and control group II （X^2 = 0-0. 337, P〈0.05）. Taking ^14 C-UBT as gold standard, the sensitivity, specificity, accuracy of the rapid testing plate were 98. 3 %, 99.2 %and 98.6 %, and that of testing strip for detec- ting HPSA were 97.4 %, 98. 4 % and 97.8 %, respectively. Conclusion The rapid testing plate of lateral-flow immunochromatographic with double antibody sandwich of colloidal gold-labeled polyclonal antibody, it is simple, inexpensive, reproduci- ble,noninvasive,and has very high sensitivity and specificity for the detection of HPSA in HP infection diagnosis.

关 键 词：幽门螺杆菌 粪便 抗原 免疫层析技术 

分 类 号：R378.2[医药卫生—病原生物学] R446[医药卫生—基础医学]