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False-positive results in a recombinant severe acute respiratory syndrome-associated coronavirus (SARS-CoV) nucleocapsid-based western blot assay were rectified by the use of two subunits (S1 and S2) of spike for detection of antibody to SARS-CoV.
Clin Vaccine Immunol. 2006 Mar; 13(3):409-14.CV

Abstract

To evaluate the reactivity of the recombinant proteins expressed in Escherichia coli strain BL21(DE3), a Western blot assay was performed by using a panel of 78 serum samples obtained, respectively, from convalescent-phase patients infected with severe acute respiratory syndrome-associated coronavirus (SARS-CoV) (30 samples) and from healthy donors (48 samples). As antigen for detection of SARS-CoV, the nucleocapsid protein (N) showed high sensitivity and strong reactivity with all samples from SARS-CoV patients and cross-reacted with all serum samples from healthy subjects, with either those obtained from China (10 samples) or those obtained from France (38 serum samples), giving then a significant rate of false positives. Specifically, our data indicated that the two subunits, S1 (residues 14 to 760) and S2 (residues 761 to 1190), resulted from the divided spike reacted with all samples from SARS-CoV patients and without any cross-reactivity with any of the healthy serum samples. Consequently, these data revealed the nonspecific nature of N protein in serodiagnosis of SARS-CoV compared with the S1 and S2, where the specificity is of 100%. Moreover, the reported results indicated that the use of one single protein as a detection antigen of SARS-CoV infection may lead to false-positive diagnosis. These may be rectified by using more than one protein for the serodiagnosis of SARS-CoV.

Authors+Show Affiliations

Emerging Pathogens Department of bioMérieux, CERVI, 21 Avenue Tony Garnier, 69365 cedex 07, Lyon, France. mmaache@ugr.esNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article

Language

eng

PubMed ID

16522785

Citation

Maache, Mimoun, et al. "False-positive Results in a Recombinant Severe Acute Respiratory Syndrome-associated Coronavirus (SARS-CoV) Nucleocapsid-based Western Blot Assay Were Rectified By the Use of Two Subunits (S1 and S2) of Spike for Detection of Antibody to SARS-CoV." Clinical and Vaccine Immunology : CVI, vol. 13, no. 3, 2006, pp. 409-14.
Maache M, Komurian-Pradel F, Rajoharison A, et al. False-positive results in a recombinant severe acute respiratory syndrome-associated coronavirus (SARS-CoV) nucleocapsid-based western blot assay were rectified by the use of two subunits (S1 and S2) of spike for detection of antibody to SARS-CoV. Clin Vaccine Immunol. 2006;13(3):409-14.
Maache, M., Komurian-Pradel, F., Rajoharison, A., Perret, M., Berland, J. L., Pouzol, S., Bagnaud, A., Duverger, B., Xu, J., Osuna, A., & Paranhos-Baccalà, G. (2006). False-positive results in a recombinant severe acute respiratory syndrome-associated coronavirus (SARS-CoV) nucleocapsid-based western blot assay were rectified by the use of two subunits (S1 and S2) of spike for detection of antibody to SARS-CoV. Clinical and Vaccine Immunology : CVI, 13(3), 409-14.
Maache M, et al. False-positive Results in a Recombinant Severe Acute Respiratory Syndrome-associated Coronavirus (SARS-CoV) Nucleocapsid-based Western Blot Assay Were Rectified By the Use of Two Subunits (S1 and S2) of Spike for Detection of Antibody to SARS-CoV. Clin Vaccine Immunol. 2006;13(3):409-14. PubMed PMID: 16522785.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - False-positive results in a recombinant severe acute respiratory syndrome-associated coronavirus (SARS-CoV) nucleocapsid-based western blot assay were rectified by the use of two subunits (S1 and S2) of spike for detection of antibody to SARS-CoV. AU - Maache,Mimoun, AU - Komurian-Pradel,Florence, AU - Rajoharison,Alain, AU - Perret,Magali, AU - Berland,Jean-Luc, AU - Pouzol,Stéphane, AU - Bagnaud,Audrey, AU - Duverger,Blandine, AU - Xu,Jianguo, AU - Osuna,Antonio, AU - Paranhos-Baccalà,Glaucia, PY - 2006/3/9/pubmed PY - 2006/5/24/medline PY - 2006/3/9/entrez SP - 409 EP - 14 JF - Clinical and vaccine immunology : CVI JO - Clin Vaccine Immunol VL - 13 IS - 3 N2 - To evaluate the reactivity of the recombinant proteins expressed in Escherichia coli strain BL21(DE3), a Western blot assay was performed by using a panel of 78 serum samples obtained, respectively, from convalescent-phase patients infected with severe acute respiratory syndrome-associated coronavirus (SARS-CoV) (30 samples) and from healthy donors (48 samples). As antigen for detection of SARS-CoV, the nucleocapsid protein (N) showed high sensitivity and strong reactivity with all samples from SARS-CoV patients and cross-reacted with all serum samples from healthy subjects, with either those obtained from China (10 samples) or those obtained from France (38 serum samples), giving then a significant rate of false positives. Specifically, our data indicated that the two subunits, S1 (residues 14 to 760) and S2 (residues 761 to 1190), resulted from the divided spike reacted with all samples from SARS-CoV patients and without any cross-reactivity with any of the healthy serum samples. Consequently, these data revealed the nonspecific nature of N protein in serodiagnosis of SARS-CoV compared with the S1 and S2, where the specificity is of 100%. Moreover, the reported results indicated that the use of one single protein as a detection antigen of SARS-CoV infection may lead to false-positive diagnosis. These may be rectified by using more than one protein for the serodiagnosis of SARS-CoV. SN - 1556-6811 UR - https://www.unboundmedicine.com/medline/citation/16522785/False_positive_results_in_a_recombinant_severe_acute_respiratory_syndrome_associated_coronavirus__SARS_CoV__nucleocapsid_based_western_blot_assay_were_rectified_by_the_use_of_two_subunits__S1_and_S2__of_spike_for_detection_of_antibody_to_SARS_CoV_ L2 - http://cvi.asm.org/cgi/pmidlookup?view=long&pmid=16522785 DB - PRIME DP - Unbound Medicine ER -