Endo F.,Tokyo Metropolitan Institute of Medical Science |
Tabata T.,Mie University |
Sadato D.,Tokyo Metropolitan Institute of Medical Science |
Sadato D.,Tokyo University of Science |
And 9 more authors.
PLoS ONE | Year: 2017
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. © 2017 Endo et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
PubMed | ADTEC Co., Meiji Pharmaceutical University, Koujin kai Kimachi Hospital, Mutsu General Hospital and 4 more.
Type: Journal Article | Journal: PloS one | Year: 2015
We developed an immunochromatography-based assay for detecting antibodies against recombinant -galactosidase A proteins in serum. The evaluation of 29 serum samples from Fabry patients, who had received enzyme replacement therapy with agalsidase alpha and/or agalsidase beta, was performed by means of this assay method, and the results clearly revealed that the patients exhibited the same level of antibodies against both agalsidase alpha and agalsidase beta, regardless of the species of recombinant -galactosidase A used for enzyme replacement therapy. A conventional enzyme-linked immunosorbent assay supported the results. Considering these, enzyme replacement therapy with agalsidase alpha or agalsidase beta would generate antibodies against the common epitopes in both agalsidase alpha and agalsidase beta. Most of the patients who showed immunopositive reaction exhibited classic Fabry phenotype and harbored gene mutations affecting biosynthesis of -galactosidase A. As immunochromatography is a handy and simple assay system which can be available at bedside, this assay method would be extremely useful for quick evaluation or first screening of serum antibodies against agalsidase alpha or agalsidase beta in Fabry disease with enzyme replacement therapy.
Sakurai A.,Tokyo Metropolitan Institute of Medical Science |
Takayama K.,ADTEC Co. |
Nomura N.,Tokyo Metropolitan Institute of Medical Science |
Munakata T.,Tokyo Metropolitan Institute of Medical Science |
And 13 more authors.
PLoS ONE | Year: 2013
Immunochromatography (IC) is an antigen-detection assay that plays an important role in the rapid diagnosis of influenza virus because the protocol is short time and easy to use. Despite the usability of IC, the sensitivity is approximately 103 pfu per reaction. In addition, antigen-antibody interaction-based method cannot be used for the detection of influenza viruses with major antigenic change. In this study, we established the use of fluorescent immunochromatography (FLIC) to detect a broad spectrum of H5 subtype influenza A viruses. This method has improved sensitivity 10-100 fold higher than traditional IC because of the use of fluorescent conjugated beads. Our Type-E FLIC kit detected all of the H5 subtype influenza viruses that were examined, as well as recombinant hemagglutinin (HA) proteins (rHAs) belonging to the Eurasian H5 subtype viruses and the Type-N diagnosed North American H5 subtype influenza A viruses. Thus, this kit has the improved potential to detect H5 subtype influenza viruses of different clades with both Type-E and Type-N FLIC kits. Compared with PCR-based diagnosis, FLIC has a strong advantage in usability, because the sample preparation required for FLIC is only mix-and-drop without any additional steps such as RNA extraction. Our results can provide new strategies against the spread and transmission of HPAI H5N1 viruses in birds and mammals including humans. Copyright: © 2013 Sakurai et al.
Adtec Inc. and Konica Minolta | Date: 2013-04-02
A method for detecting or quantifying an analyte, using a test strip for lateral flow type chromatography which contains a membrane and a detection part on which a capturing ligand that is to specifically bind to the analyte has been fixed on the membrane, includes: bringing an analyte contained in a sample into contact with a labeled ligand labeled with a phosphor that is to be excited by light having a wavelength from 600 nm to 800 nm to generate fluorescence, bringing a complex containing the analyte and the labeled ligand into contact with a capturing ligand at the detection part, and irradiating on the test strip light having a wavelength from 600 nm to 800 nm as an excitation light for the phosphor contained in the complex to generate fluorescence from the phosphor, and measuring a fluorescence intensity of the fluorescence.
Konica Minolta and Adtec Inc. | Date: 2015-02-11
To provide a method for detecting or quantifying an analyte, by which the fluorescence intensity of a fluorescence can be measured in a state where an autofluorescence from the material of a test strip is reduced or eliminated, and thereby, the sensitivity in analyte detection or quantification can be improved. The method for detecting or quantifying an analyte contained in a sample according to the present invention comprises a step of, by using the specified test strip for a lateral flow type chromatography, bringing an analyte contained in a sample into contact with a labeled ligand (1) wherein a ligand (1) that is to specifically bind to the analyte has been labeled with a phosphor (1) that is to be excited by the specified light, and the specified steps.
PubMed | ADTEC Co., Konica Minolta, Tokyo Metropolitan Institute of Medical Science and Hokkaido University
Type: Journal Article | Journal: PloS one | Year: 2015
Lateral flow tests also known as Immunochromatography (IC) is an antigen-detection method conducted on a nitrocellulose membrane that can be completed in less than 20 min. IC has been used as an important rapid test for clinical diagnosis and surveillance of influenza viruses, but the IC sensitivity is relatively low (approximately 60%) and the limit of detection (LOD) is as low as 10 pfu per reaction. Recently, we reported an improved IC assay using antibodies conjugated with fluorescent beads (fluorescent immunochromatography; FLIC) for subtyping H5 influenza viruses (FLIC-H5). Although the FLIC strip must be scanned using a fluorescent reader, the sensitivity (LOD) is significantly improved over that of conventional IC methods. In addition, the antibodies which are specific against the subtypes of influenza viruses cannot be available for the detection of other subtypes when the major antigenicity will be changed. In this study, we established the use of FLIC to type seasonal influenza A and B viruses (FLIC-AB). This method has improved sensitivity to 100-fold higher than that of conventional IC methods when we used several strains of influenza viruses. In addition, FLIC-AB demonstrated the ability to detect influenza type A and influenza type B viruses from clinical samples with high sensitivity and specificity (Type A: sensitivity 98.7% (74/75), specificity 100% (54/54), Type B: sensitivity 100% (90/90), specificity 98.2% (54/55) in nasal swab samples) in comparison to the results of qRT-PCR. And furthermore, FLIC-AB performs better in the detection of early stage infection (under 13 h) than other conventional IC methods. Our results provide new strategies to prevent the early-stage transmission of influenza viruses in humans during both seasonal outbreaks and pandemics.
Santivanez S.J.,Instituto Peruano Of Parasitologia Clinica Y Experimental |
Rodriguez M.L.,Instituto Peruano Of Parasitologia Clinica Y Experimental |
Rodriguez S.,Instituto Nacional Of Ciencias Neurologicas |
Sako Y.,Asahikawa University |
And 9 more authors.
Journal of Clinical Microbiology | Year: 2015
Diagnosis of cystic echinococcosis (CE) is based on the identification of the cyst(s) by imaging, using immunodiagnostic tests mainly as complementary tools in clinical settings. Among the antigens used for immunodiagnosis, previous studies described a good performance of the recombinant antigen B8/1 (rAgB) in an enzyme-linked immunosorbent assay (ELISA) format; however, in remote parts of areas where the disease is endemic, the implementation of an ELISA is difficult, so a more simple, rapid, and reliable method such as the immunochromatographic test (ICT) is required. In this study, using a set of 50 serum samples from patients with surgically confirmed CE, we compared the performance of an ICT and that of an ELISA using the rAgB. The overall sensitivities of ICT and ELISA were not statistically different (78% versus 72%; P=0.36). The overall agreement between both tests was moderate (κ=0.41; P<0.01). Concordance between ICT and ELISA was substantial or almost perfect for patients with liver involvement (κ=0.65; P<0.001) and patients with more than one hydatid cyst (κ=0.82; P<0.001), respectively. Moreover, specificity analysis using a total of 88 serum samples from healthy individuals (n=20) and patients (n=68) with other parasitic infections revealed that ICT had a specificity of 89.8%. ICT and ELISA had similar performance for the detection of specific antibodies to E. granulosus, and ICT had a high specificity, opening the possibility of using ICT as a screening tool in rural settings. © 2015, American Society for Microbiology.
PubMed | Instituto Nacional Of Ciencias Neurologicas, Hospital Nacional Dos Of Mayo, Adtec Co., National Major San Marcos University and 4 more.
Type: Comparative Study | Journal: Journal of clinical microbiology | Year: 2015
Diagnosis of cystic echinococcosis (CE) is based on the identification of the cyst(s) by imaging, using immunodiagnostic tests mainly as complementary tools in clinical settings. Among the antigens used for immunodiagnosis, previous studies described a good performance of the recombinant antigen B8/1 (rAgB) in an enzyme-linked immunosorbent assay (ELISA) format; however, in remote parts of areas where the disease is endemic, the implementation of an ELISA is difficult, so a more simple, rapid, and reliable method such as the immunochromatographic test (ICT) is required. In this study, using a set of 50 serum samples from patients with surgically confirmed CE, we compared the performance of an ICT and that of an ELISA using the rAgB. The overall sensitivities of ICT and ELISA were not statistically different (78% versus 72%; P = 0.36). The overall agreement between both tests was moderate ( = 0.41; P < 0.01). Concordance between ICT and ELISA was substantial or almost perfect for patients with liver involvement ( = 0.65; P < 0.001) and patients with more than one hydatid cyst ( = 0.82; P < 0.001), respectively. Moreover, specificity analysis using a total of 88 serum samples from healthy individuals (n = 20) and patients (n = 68) with other parasitic infections revealed that ICT had a specificity of 89.8%. ICT and ELISA had similar performance for the detection of specific antibodies to E. granulosus, and ICT had a high specificity, opening the possibility of using ICT as a screening tool in rural settings.
PubMed | Japan National Institute of Infectious Diseases, Khon Kaen University and Adtec Inc.
Type: | Journal: Parasites & vectors | Year: 2016
Human gnathostomiasis is a serious tropical disease, which is often overlooked. There is an urgent need to improve tools to aid the potential diagnosis of the disease in endemic regions. To overcome this, we produced the immunochromatographic test (ICT) kit for a rapid and simple diagnosis of human gnathostomiasis.The recombinant protein (named rGslic18) was applied to ICT kit as the antigen. The diagnostic value of ICT kit was evaluated using serum samples from parasitologically proven and clinically suspected gnathostomiasis patients, healthy volunteers and patients with other parasitic diseases. The ICT kit exhibited quite high sensitivity (93.75%) and specificity (97.01%).The ICT kit is simple, convenient and easy to implement and expected to provide reliable diagnostic results for human gnathostomiasis. It also will be a promising diagnostic tool not only for large-scale epidemiological surveys in endemic or remote areas where diagnostic facilities are poor but also for a rapid clinical diagnosis in the bedside laboratory.
Sako Y.,Asahikawa University |
Tappe D.,University of Würzburg |
Fukuda K.,Adtec Co. |
Kobayashi Y.,Adtec Co. |
And 5 more authors.
Clinical and Vaccine Immunology | Year: 2011
The performance of a rapid and simple immunochromatographic test (ICT) with recombinant Em18 (rEm18) antigen for serological follow-up of Echinococcus multilocularis infection was evaluated by comparison with that of an enzyme-linked immunosorbent assay (ELISA) with rEm18, using serum samples from patients who underwent surgery and/or received antiparasitic chemotherapy. The degree of Em18-band intensity on the ICT correlated highly with the absorbance value obtained by the ELISA. The kinetics of antibody levels obtained by the ICT paralleled those of the ELISA. These data suggest that the ICT has high potential as an easy-to-handle, fast, and reliable follow-up tool to monitor the status of alveolar echinococcosis in different stages. Copyright © 2011, American Society for Microbiology. All Rights Reserved.