An improved microtiter assay for evaluating anti-HIV-1 neutralizing antibodies from sera or plasma

BMC Infectious Diseases

Kunlong Ben²  Yunyun Chen³  Chiyu Zhang¹ 

[1] The School of Medical Technology, Jiangsu University, Zhenjiang, Jiangsu 212001, China;Kunming Chinawave Biotechnology Company Ltd., Kunming, Yunnan 650106, China;Laboratory for Molecular and Cell Immunology, Kunming Institute of Zoology, Chinese Academy of Sciences, Kunming, Yunnan 650223, China

关键词: serum or plasma;    microtiter neutralization assay;    MTT;    neutralizing antibody;    HIV-1;   

Others  :  1177108
DOI  :  10.1186/1471-2334-3-30

 received in 2003-10-05, accepted in 2003-12-23,  发布年份 2003

【 摘 要 】

Background

The anti-HIV-1 neutralizing antibody assay is widely used in AIDS vaccine research and other experimental and clinical studies. The vital dye staining method applied in the detection of anti-HIV-1 neutralizing antibody has been used in many laboratories. However, the unknown factor(s) in sera or plasma affected cell growth and caused protection when the tested sera or plasma was continuously maintained in cell culture. In addition, the poor solubility of neutral red in medium (such as RPMI-1640) also limited the use of this assay.

Methods

In this study, human T cell line C8166 was used as host cells, and 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) instead of neutral red was used as vital dye. In order to avoid the effect of the unknown factor(s), the tested sera or plasma was removed by a washout procedure after initial 3–6 h culture in the assay.

Result

This new assay eliminated the effect of the tested sera or plasma on cell growth, improved the reliability of detection of anti-HIV-1 neutralizing antibody, and showed excellent agreement with the p24 antigen method.

Conclusion

The results suggest that the improved assay is relatively simple, highly duplicable, cost-effective, and well reliable for evaluating anti-HIV-1 neutralizing antibodies from sera or plasma.

【 授权许可】

   
2003 Zhang et al; licensee BioMed Central Ltd. This is an Open Access article: verbatim copying and redistribution of this article are permitted in all media for any purpose, provided this notice is preserved along with the article's original URL.

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