Improving dengue viral antigens detection in dengue patient serum specimens using a low pH glycine buffer treatment.
J Microbiol Immunol Infect
; 50(2): 167-174, 2017 Apr.
Article
em En
| MEDLINE
| ID: mdl-26260863
ABSTRACT
BACKGROUND/PURPOSES:
Early diagnosis of dengue virus (DENV) infection to monitor the potential progression to hemorrhagic fever can influence the timely management of dengue-associated severe illness. Nonstructural protein 1 (NS1) antigen detection in acute serum specimens has been widely accepted as an early diagnostic assay for dengue infection; however, lower sensitivity of the NS1 antigen-capture enzyme-linked immunosorbent assay (Ag-ELISA) in secondary dengue viral infection has been reported.METHODS:
In this study, we developed two forms of Ag-ELISA capable of detecting E-Ag containing virion and virus-like particles, and secreted NS1 (sNS1) antigens, respectively. The temporal kinetics of viral RNA, sNS1, and E-Ag were evaluated based on the in vitro infection experiment. Meanwhile, a panel of 62 DENV-2 infected patients' sera was tested.RESULTS:
The sensitivity was 3.042 ng/mL and 3.840 ng/mL for sNS1 and E, respectively. The temporal kinetics of the appearance of viral RNA, E, NS1, and infectious virus in virus-infected tissue culture media suggested that viral RNAs and NS1 antigens could be detected earlier than E-Ag and infectious virus. Furthermore, a panel of 62 sera from patients infected by DENV Serotype 2 was tested. Treating clinical specimens with the dissociation buffer increased the detectable level of E from 13% to 92% and NS1 antigens from 40% to 85%.CONCLUSION:
Inclusion of a low-pH glycine buffer treatment step in the commercially available Ag-ELISA is crucial for clinical diagnosis and E-containing viral particles could be a valuable target for acute DENV diagnosis, similar to NS1 detection.Palavras-chave
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Base de dados:
MEDLINE
Assunto principal:
Ensaio de Imunoadsorção Enzimática
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Dengue
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Vírus da Dengue
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Glicina
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Antígenos Virais
Idioma:
En
Ano de publicação:
2017
Tipo de documento:
Article