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目的建立单纯疱疹病毒1型(HSV-1)药物敏感性的测定方法,并建立体外耐药病毒株。方法将HSV-1接种于乳兔肾细胞中,加入不同浓度的阿昔洛韦(ACV),培养72 h后固定、染色、清点空斑数并计算药物的半数抑制浓度(IC50),通过IC50来判断HSV-1的药物敏感性。将HSV-1在含ACV环境中连续培养9代,分别在第3、6和9代测定其IC50。结果ACV对HSV-1标准株SM44毒株的IC50为0.7μg/ml。HSV-1在含ACV的环境中连续培养3代后即产生了耐药性,第3、6和9代的IC50分别为7.2、34.2μg/ml和85.3μg/ml。结论空斑抑制试验是测定病毒药物敏感性的有效、实用的方法,建立的体外耐药病毒株可用于HSV-1的耐药性研究。
Objective To establish a method for the determination of herpes simplex virus type 1 (HSV-1) drug sensitivity and to establish an in vitro drug-resistant virus strain. Methods HSV-1 cells were inoculated into rabbit renal grafts and acyclovir (ACV) of different concentrations were added. After being cultured for 72 h, HSV-1 cells were fixed, stained, counted for plaque count and the IC50 value was calculated. To determine the drug sensitivity of HSV-1. HSV-1 was cultured continuously for 9 passages in an ACV-containing environment and its IC50 was measured at the 3rd, 6th and 9th generation respectively. Results The IC50 of ACV against HSV-1 Strain SM44 was 0.7 μg / ml. HSV-1 developed resistance after 3 generations of continuous culture in an ACV-containing environment with IC50 of 7.2, 34.2 and 85.3 μg / ml for the 3rd, 6th and 9th generation, respectively. Conclusion The plaque inhibition test is an effective and practical method for the determination of the drug sensitivity of the virus. The established drug-resistant virus strains can be used for the drug resistance study of HSV-1.