Development of Monoclonal Antibodies against HIV-1 p24 Protein and Its Application in Colloidal Gold Immunochromatographic Assay for HIV-1 Detection

Human immunodeficiency virus type 1 (HIV-1) p24 protein is the most abundant viral protein of HIV-1. This protein is secreted in blood serum at high levels during the early stages of HIV-1 infection, making it a biomarker for early diagnosis. In this study, a colloidal gold immunochromatographic assay (GICA) was established for detecting p24 protein using mouse monoclonal antibodies (mAbs). The HIV-1 p24 protein was expressed in E. coli strain BL21 and the purified protein was used to immunize mice. Stable hybridoma cell lines secreting anti-p24 monoclonal antibodies were obtained after ELISA screening and subcloning by limiting dilution. 34 different capture and labeling mAb pairs were selected by a novel antibody-capture indirect sandwich ELISA and then applied in GICA to detect p24 protein. The GICA method has a limit of detection (LOD) of 25 pg/mL and could detect p24 protein in all 10 positive samples obtained from the National Reference of HIV-1 p24 antigen. Out of 153 negative samples tested, 3 false positives results were obtained. The overall specificity of this test was 98.03%. The good sensitivity and specificity of this method make it a suitable alternative to provide a more convenient and efficient tool for early diagnosis of HIV infection

Rapid Diagnostic Test and Real Time RT-PCR of clinical samples from influenza A(H7N9) positive patients.

<p>Rapid Diagnostic Test and Real Time RT-PCR of clinical samples from influenza A(H7N9) positive patients.</p

Rapid Diagnostic Test Quality Control Panel.

<p>* Signal intensity was determined by comparing with a standard color chart and categorized as weak (+, C7–9), moderate (++, C4–6) or strong (+++, C1–3) positivity, respectively. “−” indicates a negative test result.</p

Analytic sensitivity and specificity of rapid diagnostic test for recombinant hemagglutinin derived from influenza viruses.

<p>*Signal intensity was determined by comparing with a standard color chart and categorized as weak (+, C7–9), moderate (++, C4–6) or strong (+++, C1–3) positivity, respectively. “−” indicates a negative test result.</p

Detection Limit of Rapid Diagnostic Test and Real Time RT-PCR for H7N9 isolates.

a<p>State Key Laboratory for Diagnosis and Treatment of Infection Diseases, First Affiliated Hospital, College of Medicine, Zhejiang University.</p>b<p>Beijing #1: Influenza Branch of the Center for Disease Prevention and Control of China.</p>c<p>Beijing #2: Institute of Microbiology and Epidemiology, Academy of Military Medical Sciences (Beijing, China).</p

Rapid Diagnostic Test of clinical samples from influenza A(H7N9) positive patients.

<p>Rapid Diagnostic Test of clinical samples from influenza A(H7N9) positive patients.</p