Nonstructural Protein 1-Specific Immunoglobulin M and G Antibody Capture Enzyme-Linked Immunosorbent Assays in Diagnosis of Flaviviral Infections in Humans

被引：24

作者：

Chao, Day-Yu ^{[1
]}

Galula, Jedhan Ucat ^{[2
]}

Shen, Wen-Fan ^{[3
]}

Davis, Brent S. ^{[4
]}

Chang, Gwong-Jen J. ^{[4
]}

机构：

[1] Natl Chung Hsing Univ, Coll Vet Med, Grad Inst Microbiol & Publ Hlth, Taichung 40227, Taiwan

[2] Natl Chung Hsing Univ, Coll Vet Med, Dept Vet Med, Taichung 40227, Taiwan

[3] Natl Chung Hsing Univ, PhD Program Microbial Genom, Taichung 40227, Taiwan

[4] Ctr Dis Control & Prevent, Div Vector Borne Dis, Ft Collins, CO USA

来源：

JOURNAL OF CLINICAL MICROBIOLOGY | 2015年 / 53卷 / 02期

关键词：

JAPANESE ENCEPHALITIS-VIRUS; GLYCOPROTEIN NS1; M IGM; DENGUE; IMMUNIZATION; ANTIGEN; FEVER; ELISA; PROTECTION; ENVELOPE;

D O I：

10.1128/JCM.02735-14

中图分类号：

Q93 [微生物学];

学科分类号：

071005 ; 100705 ;

摘要：

IgM antibody-and IgG antibody-capture enzyme-linked immunosorbent assays (MAC/GAC-ELISAs) targeted at envelope protein (E) of dengue viruses (DENV), West Nile virus, and Japanese encephalitis virus (JEV) are widely used as serodiagnostic tests for presumptive confirmation of viral infection. Antibodies directed against the flavivirus nonstructural protein 1 (NS1) have been proposed as serological markers of natural infections among vaccinated populations. The aim of the current study is to optimize an IgM and IgG antibody-capture ELISA (MAC/GAC-ELISA) to detect anti-NS1 antibodies and compare it with anti-E MAC/GAC-ELISA. Plasmids to express premembrane/envelope (prM/E) or NS1 proteins of six medically important flaviviruses, including dengue viruses (DENV-1 to DENV-4), West Nile virus (WNV), and Japanese encephalitis virus (JEV), were constructed. These plasmids were used for the production of prM/E-containing virus-like particles (VLPs) and secreted NS1 (sNS1) from COS-1 cells. Archived clinical specimens from patients with confirmed DENV, JEV, and WNV infections, along with naive sera, were subjected to NS1-MAC/GAC-ELISAs before or after depletion of anti-prM/E antibodies by preabsorption with or without VLPs. Human serum specimens from previously confirmed DENV infections showed significantly enhanced positive-to-negative (P/N) ratios for NS1-MAC/GAC-ELISAs after the depletion of anti-prM/E antibodies. No statistical differences in sensitivities and specificities were found between the newly developed NS1-and VLP-MAC/GAC-ELISAs. Further application of the assays to WNV-and JEV-infected serum panels showed similar results. A novel approach to perform MAC/GAC-ELISAs for NS1 antibody detection was successfully developed with great potential to differentiate antibodies elicited by the tetravalent chimeric yellow fever-17D/dengue vaccine or DENV infection.

引用

页码：557 / 566

页数：10

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