MTT法在抗HIV药物研究中的应用

Apply a MTT assay to studying on anti HIV drugs

目的参考多篇文献建立用于体外药物抗ＨＩＶ研究的ＭＴＴ试验方法。方法其过程可简述为先用９６孔微量板做抗病毒实验，４天后，加ＭＴＴ（０．７ｍｇ／ｍｌ）２０μｌ／孔，置３７℃ＣＯ２箱中４ｈ，移去１５０μｌ／孔培养液，再加１００μｌ／孔的ＤＭＳＯ过１０分钟，即可测定ＯＤ５４０值，计算药物对ＨＩＶ感染细胞的保护率。结果ＨＬ的ＣＣ５０＝０．３３５４ｍｇ／ｍｌ，ＥＣ５０＝０．０８６４ｍｇ／ｍｌ，ＭＴＴ法较ＣＰＥ法更为敏感。结论ＭＴＴ法能够以光密度值快速、客观、准确地得到试验结果。

Objective\ A MTT method for evaluate the efficacy of drugs for anti HIV 1 infection in vitro,was developed with reference materials.Methods\ Briefly,Anti HIV assay was first in progress with 96 well plastic microtiter trays.Four days after infection,20μl/well of MTT(0.7mg/ml) was added,the trays were incubated at 37℃ in a CO 2 incubator for 4h.150μl/well of medium was removed,then 100μl/well of DMSO was added.The absorbance (OD540) were read,then the percent protection of drugs in HIV infected cells was calculated.Results\ CC50=0.3354mg/ml,EC50=0.0864mg/ml of HL,MTT assay is more sensitive then CPE assay.Conclusion\ By the MTT assay,a rapid,objective,accurate assay results can achieve with absorbance,therefore,enhance quality and efficiency of the experiments.