中华实验和临床病毒学杂志
中華實驗和臨床病毒學雜誌
중화실험화림상병독학잡지
CHINESE JOURNAL OF EXPERIMENTAL AND CLINICAL VIROLOGY
2011年
1期
73-75
,共3页
种辉辉%许四宏%李敬云%王佑春
種輝輝%許四宏%李敬雲%王祐春
충휘휘%허사굉%리경운%왕우춘
HIV%重组假病毒法%逆转录酶抑制剂%茚地那韦%奈维拉平
HIV%重組假病毒法%逆轉錄酶抑製劑%茚地那韋%奈維拉平
HIV%중조가병독법%역전록매억제제%인지나위%내유랍평
HIV%Recombinant virus assay%Reverse transcriptase inhibitors%IDV%NVP
目的 对比重组假病毒法与活病毒法在评价抗HIV-1药物中的效果.方法 将pSG3△env质粒和带有ENV基因的质粒B01共转染获得重组假病毒.用重组假病毒分别与不同稀释度的药物共孵育,根据相对荧光单位的不同分析计算EC50、ED50.在P3实验室用活病毒分别与不同稀释度的药物共孵育,根据细胞病变程度的不同分析计算EC50.结果 用重组假病毒法和活病毒法分别检测抗HIV-1药物奈维拉平(NVP),其EC50分别是88.9 nmol/L、89.5 nmol/L,CV值分别是0和0.6%.用重组假病毒法和活病毒法分别检测抗HIV-1药物茚地那韦(IDV),其EC50分别是0.36 μmol/L、0.23μmol/L,CV值分别是0和60%.用重组假病毒法检测抗HIV-1药物可以定量计算出ED50,IDV和NVP的ED50分别是70.6 nmol/L和0.62 μmol/L,CV值分别是14.3%和9.7%.结论 重组假病毒法可以用于对抗HIV-1药物的评价.计算EC50时,重组假病毒法与活病毒法的结果 相差不超过2倍,但活病毒具有生物危害,而重组假病毒法的重复性较好,并且还可以定量计算出药物的ED50.
目的 對比重組假病毒法與活病毒法在評價抗HIV-1藥物中的效果.方法 將pSG3△env質粒和帶有ENV基因的質粒B01共轉染穫得重組假病毒.用重組假病毒分彆與不同稀釋度的藥物共孵育,根據相對熒光單位的不同分析計算EC50、ED50.在P3實驗室用活病毒分彆與不同稀釋度的藥物共孵育,根據細胞病變程度的不同分析計算EC50.結果 用重組假病毒法和活病毒法分彆檢測抗HIV-1藥物奈維拉平(NVP),其EC50分彆是88.9 nmol/L、89.5 nmol/L,CV值分彆是0和0.6%.用重組假病毒法和活病毒法分彆檢測抗HIV-1藥物茚地那韋(IDV),其EC50分彆是0.36 μmol/L、0.23μmol/L,CV值分彆是0和60%.用重組假病毒法檢測抗HIV-1藥物可以定量計算齣ED50,IDV和NVP的ED50分彆是70.6 nmol/L和0.62 μmol/L,CV值分彆是14.3%和9.7%.結論 重組假病毒法可以用于對抗HIV-1藥物的評價.計算EC50時,重組假病毒法與活病毒法的結果 相差不超過2倍,但活病毒具有生物危害,而重組假病毒法的重複性較好,併且還可以定量計算齣藥物的ED50.
목적 대비중조가병독법여활병독법재평개항HIV-1약물중적효과.방법 장pSG3△env질립화대유ENV기인적질립B01공전염획득중조가병독.용중조가병독분별여불동희석도적약물공부육,근거상대형광단위적불동분석계산EC50、ED50.재P3실험실용활병독분별여불동희석도적약물공부육,근거세포병변정도적불동분석계산EC50.결과 용중조가병독법화활병독법분별검측항HIV-1약물내유랍평(NVP),기EC50분별시88.9 nmol/L、89.5 nmol/L,CV치분별시0화0.6%.용중조가병독법화활병독법분별검측항HIV-1약물인지나위(IDV),기EC50분별시0.36 μmol/L、0.23μmol/L,CV치분별시0화60%.용중조가병독법검측항HIV-1약물가이정량계산출ED50,IDV화NVP적ED50분별시70.6 nmol/L화0.62 μmol/L,CV치분별시14.3%화9.7%.결론 중조가병독법가이용우대항HIV-1약물적평개.계산EC50시,중조가병독법여활병독법적결과 상차불초과2배,단활병독구유생물위해,이중조가병독법적중복성교호,병차환가이정량계산출약물적ED50.
Objective To compaire results of recombinant virus assay and live virus assay on evaluateing anti-HIV-1 drugs. Methods The pseudoviruse was generated by cotransfection of the plasmid B01 containing gp160 genes and pSG3△ env plasmid. After co-incubation of pseudovirus with serially diluted drug, the EC50 and ED50 were calculated according to RLU(relative light unit) for each drug. After coincubation of live virus with serially diluted drug, the EC50 was calculated according to cytopathic effect.Results EC50 of IDV measured by the recombinant virus assay and live virus assay was 88.9 nmol/L, 89.5 nmol/L, respectively, while EC50 of NVP measured by the recombinant virus assay and live virus assay was 0. 36 μmol/L,0. 23 μmol/L, respectively. The recombinant virus assay showed good reproducibility with coefficient variation of 0, however coefficient variation of live virus assay reached to 60%. ED50 of IDV and NVP measured by the recombinant virus assay were 70.6 nmol/L and 0.62 μmol/L, respectively.Coefficient variations for IDV and NVP were 14.3% and 9.7%, respectively. Conclusion The pseudoviruses could be used in evaluating anti-HIV-1 drugs. The recombinant virus assay showed good reproducibility and could calculate not only the EC50 but also the ED50 of drugs.
