Establishment of a reference panel for the detection of anti-SARS-CoV antibodies.
The immunological assays for detection of antibodies against SARS-CoV were developed in-house and some of them are available commercially. However, the antigens used in these assays differed. In order to validate the reliability of these assays, the standard panel should be established. In this study, we have expressed and purified severe acute respiratory syndrome (SARS) structural proteins and their fragments and developed indirect enzyme-linked immunosorbent assays (ELISAs) that detect antibodies against the SARS N, N(1), N(2), S(1), S(C), S(2), and M proteins as well as the human coronavirus OC43 and 229E N proteins. These assays were used to screen 58 samples from SARS convalescent patients, 40 serial serum specimens from patients at different phases of SARS infection, and 88 plasma specimens from normal blood donors. The samples from normal blood donors were also tested for antibodies against other respiratory virus. The representative samples were chosen to comprise a reference panel of SARS antibodies that may be used for the detection of SARS. The panel is composed of 25 positive samples, 25 negative samples, 7 diluted samples for anti-N antibody, 6 diluted samples for anti-S antibody, and one sample for validating precision. Comparison of detection results with different SARS antibody assays indicated that our panel should differentiate the specificity and sensitivity of different assays.
Liu
Chunyu
C
Department of Cell Biology, National Institute for the Control of Pharmaceutical and Biological Products, No. 2 Tiantanxili, Chongwen District, Beijing 100050, PR China.
Li
Xiuhua
X
Zhang
Chuntao
C
Xu
Sihong
S
Shao
Yiming
Y
Zhuang
Hui
H
Che
Xiaoyan
X
Qiu
Yan
Y
Yin
Hongzhang
H
Li
Defu
D
Wang
Youchun
Y
D000914:Antibodies, Viral / Q000097:blood*; D000956:Antigens, Viral / Q000097:blood; D001483:Base Sequence; D000086462:Coronavirus Nucleocapsid Proteins; D004279:DNA, Viral / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000592:standards; D005814:Genes, Viral; D006801:Humans; D008562:Membrane Glycoproteins / Q000235:genetics / Q000276:immunology; D019590:Nucleocapsid Proteins / Q000235:genetics / Q000276:immunology; D012015:Reference Standards; D045473:Severe acute respiratory syndrome-related coronavirus / Q000235:genetics / Q000276:immunology*; D045169:Severe Acute Respiratory Syndrome / Q000175:diagnosis / Q000276:immunology / Q000821:virology; D064370:Spike Glycoprotein, Coronavirus; D014759:Viral Envelope Proteins / Q000235:genetics / Q000276:immunology
D000914:Antibodies, Viral; D000956:Antigens, Viral; D000086462:Coronavirus Nucleocapsid Proteins; D004279:DNA, Viral; D008562:Membrane Glycoproteins; D019590:Nucleocapsid Proteins; D064370:Spike Glycoprotein, Coronavirus; D014759:Viral Envelope Proteins