Development of a simple and quick immunochromatography method for detection of anti-HPV-16/-18 antibodies.
Development of a simple and quick immunochromatography method for detection of anti-HPV-16/-18 antibodies.
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DOI：
10.1371/journal.pone.0171314
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发表时间：
2017
期刊：
影响因子：
3.7
通讯作者：
Shibasaki F
中科院分区：
文献类型：
--
作者：
Endo F;Tabata T;Sadato D;Kawamura M;Ando N;Oboki K;Ukaji M;Kobayashi K;Kobayashi Y;Ikeda T;Shibasaki F
免疫层析（IC）广泛用于检测生物液体中的目标分子。由于该方法无需特殊技术或设备即可进行，因此 IC 是一种简单快速地评估抗体或病毒和细菌等病原体是否存在的便捷方法。在本研究中，我们建立了一种 IC 方法，以蚕产生的重组 L1 蛋白为抗原，检测针对致癌人乳头瘤病毒 (HPV)-16 和 HPV-18 L1 蛋白的血清抗体。致癌性 HPV 感染是宫颈癌的主要危险因素，宫颈癌是全世界女性最常见的癌症之一。我们首先采用磁珠酶联免疫吸附试验（MB-ELISA）测定两组的血清。对于第一组，前瞻性地从计划接受 HPV 疫苗接种的年轻女性身上收集血清。第二组包括20岁以下儿童、未接种疫苗的健康女性、接种疫苗的健康女性、不典型增生、宫颈上皮内瘤变III和宫颈癌患者。我们证实，标准疫苗接种剂量显着提高了血清HPV抗体浓度，并且该水平在疫苗接种后至少持续30个月以上。相比之下，在宫颈癌前病变和宫颈癌患者中没有观察到抗体浓度增加。接下来我们用我们最初开发的IC方法对两组样品进行了测量，发现IC的测量值与MB-ELISA的测量值高度相关。简单快速的 IC 方法将成为在非实验室环境中快速监测 L1 特异性抗体水平的有用工具。只需不到一滴血清，我们的 IC 即可在 15 分钟内轻松检测血清 HPV-16/-18 抗体，无需电子设备或技术。
Immunochromatography (IC) is widely used to detect target molecules in biological fluids. Since this method can be performed without a special technique or device, IC is a convenient way to assess the existence of antibodies or pathogens such as viruses and bacteria, simply and quickly. In this study, we established an IC method to detect serum antibodies against oncogenic human papillomavirus (HPV)-16 and HPV-18 L1 proteins using recombinant L1 proteins produced by silkworms as antigens. Infection of oncogenic HPVs is a major risk factor of cervical cancer, which is one of the most common cancers in women worldwide. We first measured blood sera of two groups by magnetic beads enzyme-linked immunosorbent assay (MB-ELISA). For the first group, sera were collected prospectively from young women who planned to receive HPV vaccination. The second group consisted of children under 20 years of age, non-vaccinated healthy women, vaccinated healthy women, dysplasia, cervical intraepithelial neoplasia III, and cervical cancer patients. We confirmed that standard vaccination doses significantly increased serum HPV antibody concentrations, and the level was sustained at least more than 30 months after vaccination. In contrast, an increase in antibody concentration was not observed in patients with precancerous cervical changes and cervical cancer. We next measured the samples in both groups using the IC method we originally developed, and found that the measurement values of IC highly correlated with those of MB-ELISA. The simple and quick IC method would be a useful tool for rapid monitoring of L1 specific antibody levels in a non-laboratory environment. With less than one drop of serum, our IC can easily detect serum HPV-16/-18 antibodies within 15 minutes, without the need for electronic devices or techniques.