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4. Potential Cross-Reactivity of Severe Acute Respiratory Syndrome-Associated Coronavirus (SARS-CoV) Nucleocapsid (N)-based IgG ELISA Assay for Plasma Samples from HIV-1 Positive Intravenous Drug Usets (IDUs)

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Aim: to evaluate the specificity of the SARS-CoV N protein-based IgG ELISA assay for detection of immunoglobulin G (IgG) in plasma samples obtained from HIV-1 positive and HIV-1 negative intravenous drug users (IDUs). Methods: the SARS-Co V N gene was cloned into pQE-80L vector, and the constructs were transformed into Escherichia coli BL21. The 6 x His-tagged N protein was expressed by inducing the bacterial cells with isopropyl-1-thio-D-galactopyranoside (IPTG) and purified by Ni-NTA affinity resin. The 6 x His-tagged N protein was used as antigen for ELISA assay and evaluated for the serum samples from patients with SARS positive and the plasma samples from the HIV-I positive and negative IDUs.
Results: all sera samples from patients with SARS positive were the ELISA positive (100% sensitivity). The ELISA assay yielded no positive results of the total 61 HIV-1 negative IDU samples (100% specificity) and two positive results of the total 68 HIV-1 positive IDU samples (97.06% specificity).
Conclusion: the specificity of the SARS-CoV N protein-based IgG ELISA assay for the detection of the SARS-CoV N specific IgG in plasma samples from IDUs with HIV-1 positive is, therefore, questionable.
Key words: SARS-Co V; N protein, ELISA, intravenous drug users, HIV-1.

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Acta Medica Indonesiana: The Indonesian Journal of

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Jakarta : FK universitas indonesia., 2012

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6p

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0125-9326

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