Multiplex diagnosis of viral infectious diseases (AIDS, hepatitis C, and hepatitis A) based on point of care lateral flow assay using engineered proteinticles.
Lateral flow assay (LFA) is an attractive method for rapid, simple, and cost-effective point of care diagnosis. For LFA-based multiplex diagnosis of three viral intractable diseases (acquired immune deficiency syndrome and hepatitis C and A), here we developed proteinticle-based 7 different 3D probes that display different viral antigens on their surface, which were synthesized in Escherichia coli by self-assembly of human ferritin heavy chain that was already engineered by genetically linking viral antigens to its C-terminus. Each of the three test lines on LFA strip contains the proteinticle probes to detect disease-specific anti-viral antibodies. Compared to peptide probes, the proteinticle probes were evidently more sensitive, and the proteinticle probe-based LFA successfully diagnosed all the 20 patient sera per each disease without a false negative signal, whereas the diagnostic sensitivities in the peptide probe-based LFAs were 65-90%. Duplex and triplex assays performed with randomly mixed patient sera gave only true positive signals for all the 20 serum mixtures without any false positive signals, indicating 100% sensitivity and 100% specificity. It seems that on the proteinticle surface the antigenic peptides have homogeneous orientation and conformation without inter-peptide clustering and hence lead to the enhanced diagnostic performance with solving the problems of traditional diagnostic probes. Although the multiplex diagnosis of three viral diseases above was demonstrated as proof-of-concept here, the proposed LFA system can be applied to multiplex point of care diagnosis of other intractable diseases.
Department of Chemical and Biological Engineering, College of Engineering, Korea University, Anam-Ro 145, Seoul 136-713, Republic of Korea.
Jong-Hwan
JH
Lee
Department of Chemical and Biological Engineering, College of Engineering, Korea University, Anam-Ro 145, Seoul 136-713, Republic of Korea.
Hyuk Seong
HS
Seo
Department of Chemical and Biological Engineering, College of Engineering, Korea University, Anam-Ro 145, Seoul 136-713, Republic of Korea.
Jung-Hyuk
JH
Kwon
Department of Chemical and Biological Engineering, College of Engineering, Korea University, Anam-Ro 145, Seoul 136-713, Republic of Korea.
Hee-Tae
HT
Kim
Department of Chemical and Biological Engineering, College of Engineering, Korea University, Anam-Ro 145, Seoul 136-713, Republic of Korea.
Koo Chul
KC
Kwon
Department of Chemical and Biological Engineering, College of Engineering, Korea University, Anam-Ro 145, Seoul 136-713, Republic of Korea.
Sang Jun
SJ
Sim
College of Medicine, Chung-Ang University, Heuksuk-Ro 84, Seoul 156-756, Republic of Korea.
Young Joo
YJ
Cha
Department of Chemical and Biological Engineering, College of Engineering, Korea University, Anam-Ro 145, Seoul 136-713, Republic of Korea. Electronic address: leejw@korea.ac.kr.
Jeewon
J
Lee
D000163:Acquired Immunodeficiency Syndrome / Q000097:blood / Q000175:diagnosis* / Q000276:immunology; D000956:Antigens, Viral / Q000235:genetics / Q000276:immunology; D004867:Equipment Design; D019544:Equipment Failure Analysis; D006506:Hepatitis A / Q000097:blood / Q000175:diagnosis* / Q000276:immunology; D006526:Hepatitis C / Q000097:blood / Q000175:diagnosis* / Q000276:immunology; D006801:Humans; D007118:Immunoassay / Q000295:instrumentation; D056656:Lab-On-A-Chip Devices; D000067716:Point-of-Care Testing; D015202:Protein Engineering / Q000379:methods; D011934:Reagent Strips; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D019562:Viral Load / Q000295:instrumentation
D000956:Antigens, Viral; D011934:Reagent Strips
Infectious diseases; Lateral flow assay; Multiplex diagnosis; Proteinticles; Surface engineering