Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2004 Mar;11(2):287-91.
doi: 10.1128/cdli.11.2.287-291.2004.

Recombinant protein-based enzyme-linked immunosorbent assay and immunochromatographic tests for detection of immunoglobulin G antibodies to severe acute respiratory syndrome (SARS) coronavirus in SARS patients

Ming Guan  1 , Hsiao Ying Chen, Shen Yun Foo, Yee-Joo Tan, Phuay-Yee Goh, Shock Hwa Wee
Affiliations

Recombinant protein-based enzyme-linked immunosorbent assay and immunochromatographic tests for detection of immunoglobulin G antibodies to severe acute respiratory syndrome (SARS) coronavirus in SARS patients

Ming Guan et al. Clin Diagn Lab Immunol. 2004 Mar.

Abstract

An enzyme-linked immunosorbent assay (ELISA) and a rapid immunochromatographic test for detection of immunoglobulin G (IgG) antibodies in severe acute respiratory syndrome (SARS) patients were developed by utilizing the well-characterized recombinant proteins Gst-N and Gst-U274. The ELISA detected IgG antibodies to SARS-CoV in all 74 convalescent-phase samples from SARS patients while weakly cross-reacting to only 1 of the 210 control sera from healthy donors. This finding thus led to a kit sensitivity, specificity, and accuracy of 100, 99.5, and 99.6%, respectively. The test thus provided a positive predictive value (PPV) of 98.7% and a negative predictive value (NPV) of 100%. In addition, the ELISA gave a positive delta of 5.4 and a negative delta of 3.6, indicating an excellent differentiation between positives and negatives. The same recombinant proteins were also applied to a newly developed platform for the development of a 15-min rapid test. The resulting rapid test has an excellent agreement of 99.6%, with a kappa value of 1.00, with the ELISA. Again, this rapid test was able to detect 100% of the samples tested (n = 42) while maintaining a specificity of 99.0% (n = 210). The PPV and NPV for the rapid test thus reached 95.3 and 100%, respectively.

PubMed Disclaimer

Figures

FIG. 1.
FIG. 1.
Examples of the assembled rapid immunochromatographic test devices with their separators (transparent tabs) at the “removed” (after assay) position. To the left is a device after an assay with a sample from a patient, and to the right is another after an assay with a sample from a healthy control. The three lines in the viewing window for the positive sample represent (from top to bottom) the control, Gst-N, and Gst-U274, respectively. The negative sample generated the control line only.
FIG. 2.
FIG. 2.
Scatter chart of OD values obtained with sera from both SARS patients and healthy controls tested for IgG antibody to SARS-CoV by using the newly developed ELISA.
FIG. 3.
FIG. 3.
Titration curves of seven SARS patient samples obtained with the ELISA.
FIG. 4.
FIG. 4.
Correlation between the ELISA and the rapid test when comparing the dilutions at which reactivity end points were obtained with the seven SARS patient samples.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Centers for Disease Control and Prevention. 2003. Updated interim surveillance case definition for severe acute respiratory syndrome (SARS)—United States, April 29, 2003. Morb. Mortal. Wkly. Rep. 52:1-3. - PubMed
    1. Crofts, N., W. Maskill, and D. Gust. 1988. Evaluation of enzyme-linked immunosorbent assays: a method of data analysis. J. Virol. Methods 22:51-59. - PubMed
    1. Drosten, C., S. Gunther, W. Preiser, S. van der Werf, H. R. Brot, S. Becker, H. Rabenau, M. Panning, L. Kolesnikova, R. A. Fouchier, A. Berger, A. M. Burguiere, J. Cinatl, M. Eickmann, N. Escriou, K Grywna, S. Kramme, J. C. Manuguerra, S. Muller, V. Rickerts, M. Sturmer, S. Vieth, H. D. Klenk, A. D. Osterhaus, H. Schmitz, and H. W. Doerr. 2003. Identification of a novel coronavirus in patients with severe acute respiratory syndrome. N. Engl. J. Med. 384:1967-1976. - PubMed
    1. Feng, P., S. H. Chan, M. Y. R. Soo, D. X. Liu, M. Guan, E. R. Ren, and H. Z. Hu. 2001. Antibody response to Epstein-Barr virus Rta protein in patients with nasopharyngeal carcinoma. Cancer 92:1872-1880. - PubMed
    1. Fouchier, R. A. M., T. Kuiken, M. Schutten, G. van Amerongen, G. J. J. van Doornum, B. G. van den Hoogen, M. Peiris, W. Lim, K. Stohr, and A. D. M. E. Osterhaus. 2003. Aetiology: Koch's postulates fulfilled for SARS virus. Nature 423:240. - PMC - PubMed

Publication types

MeSH terms