Development of a sensitive immunochromatographic kit using fluorescent silica nanoparticles for rapid serodiagnosis of amebiasis
Development of a sensitive immunochromatographic kit using fluorescent silica nanoparticles for rapid serodiagnosis of amebiasis
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使用荧光二氧化硅纳米颗粒开发灵敏的免疫色谱试剂盒，用于阿米巴病的快速血清诊断
DOI：
10.1017/s0031182018000690
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发表时间：
2018
期刊：
影响因子：
2.4
通讯作者：
Masuda Gohta
中科院分区：
文献类型：
--
作者：
Tachibana Hiroshi;Kakino Azumi;Kazama Makoto;Feng Meng;Asai Satomi;Umezawa Kazuo;Nozaki Tsukasa;Makiuchi Takashi;Kamada Takayuki;Watanabe Hideki;Horiki Noriyuki;Cheng Xunjia;Masuda Gohta
我们以前已经表明，溶组织内阿米巴半乳糖-和N-乙酰-D-半乳糖胺-凝集素（C-Igl）的中间亚基的C-末端区域是一个有用的抗原阿米巴病的血清学诊断。利用在大肠杆菌中制备的C-Igl包被的荧光二氧化硅纳米颗粒开发了免疫层析试剂盒。将用于检查的样品加入到冷冻干燥的颗粒中，然后施加到免疫色谱装置上，其中膜上的测试线也用C-Igl包被。使用手持式阅读器测量荧光强度。在使用人单克隆抗体对试剂盒进行评价时，显示阳性结果的C-Igl特异性抗体的最低量为100微克。在对试剂盒间接免疫荧光抗体试验中不同抗体滴度的血清样本进行评价时，20 µL血清足以在30 min时获得阳性结果。无症状E.溶组织囊肿携带者试剂盒检测结果为阳性。用健康人和其他感染性疾病患者的血清进行评价，该试剂盒的敏感性和特异性分别为100%和97.6%。因此，我们认为，新开发的试剂盒是有用的快速血清学诊断阿米巴病。
We have previously shown that the C-terminal region of the intermediate subunit of Entamoeba histolytica galactose- and N-acetyl-D-galactosamine-inhibitable lectin (C-Igl) is a useful antigen for serodiagnosis of amebiasis. An immunochromatographic kit was developed using fluorescent silica nanoparticles coated with C-Igl prepared in Escherichia coli. Samples for examination were added to the freeze-dried particles and then applied to the immunochromatographic device, in which a test line on the membrane was also coated with C-Igl. Fluorescent intensity was measured using a hand-held reader. In an evaluation of the kit using a human monoclonal antibody, the minimum amount of C-Igl specific antibody showing positive results was 100 pg. In the evaluation of serum samples with different antibody titers in indirect immunofluorescent antibody tests in the kit, 20 µL of serum was sufficient to obtain positive results at 30 min. Serum samples from symptomatic patients with amebic colitis and amebic liver abscess and those from asymptomatic E. histolytica-cyst carriers showed positive results in the kit. Based on evaluation using sera from healthy controls and patients with other infectious diseases, the sensitivity and specificity of the kit were 100 and 97.6%, respectively. Therefore, we conclude that the newly developed kit is useful for rapid serodiagnosis of amebiasis.