Novel rapid immunochromatographic test based on an enzyme immunoassay for detecting nucleocapsid antigen in SARS-associated coronavirus
| Autor: | Hiroyuki Kogaki, Yoshihiro Kurano, Yoshiaki Uchida, Ikuo Takashima, Kazushige Miyake, Hiko Tamashiro, Yasuji Kido, Hiroaki Kariwa, Masahisa Okada, Nobuyuki Fujii, Ai Ee Ling |
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| Rok vydání: | 2005 |
| Předmět: |
Microbiology (medical)
Time Factors medicine.drug_class Clinical Biochemistry DNA Recombinant Severe Acute Respiratory Syndrome Monoclonal antibody medicine.disease_cause Sensitivity and Specificity law.invention Immunoenzyme Techniques Mice Antigen law Chlorocebus aethiops Escherichia coli medicine Animals Immunology and Allergy Nucleocapsid Vero Cells Coronavirus Biochemistry medical Chromatography biology medicine.diagnostic_test fungi Biochemistry (medical) Public Health Environmental and Occupational Health Antibodies Monoclonal Reproducibility of Results Original Articles Hematology Virology Molecular biology Medical Laboratory Technology Immunoassay Vero cell biology.protein Recombinant DNA Antibody |
| Zdroj: | Journal of Clinical Laboratory Analysis |
| ISSN: | 1098-2825 0887-8013 |
| DOI: | 10.1002/jcla.20070 |
| Popis: | A novel severe acute respiratory syndrome‐associated coronavirus (SARS‐CoV) has been discovered. The detection of both antigens and antibodies in SARS‐CoV from human specimens with suspected SARS plays an important role in preventing infection. We developed a novel rapid immunochromatographic test (RICT) based on the sandwich format enzyme immunoassay (EIA) with an all‐in‐one device for detecting the native nucleocapsid antigen (N‐Ag) of SARS‐CoV using monoclonal antibodies (MoAbs), which we produced by immunizing recombinant N‐Ag to mice. RICT is a qualitative assay for respiratory aspirates and serum specimens. With this assay, a positive result can be judged subjectively by the appearance of a blue line on the device 15 min after the sample is applied. RICT with several pairs of MoAbs showed a high sensitivity for the detection of recombinant N‐Ag as well as viral N‐Ag of SARS‐CoV. rSN122 and rSN21‐2 were the best MoAbs for immobilized antibody and enzyme labeling, respectively. With regard to analytical sensitivity, RICT detected N‐Ag at 31 pg/mL for recombinant N‐Ag, and at 1.99×10(2) TCID(50)/mL for SARS‐CoV. The specificity of RICT was 100% when 150 human sera and 50 nasopharyngeal aspirates (NSPs) were used. RICT based on an EIA using the rSN122/rSN21‐2 pair is a sensitive, specific, and reliable rapid assay for detecting N‐Ag in SARS‐CoV treated with either heat or Triton X‐100. J. Clin. Lab. Anal. 19:150–159, 2005. © 2005 Wiley‐Liss, Inc. |
| Databáze: | OpenAIRE |
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