单纯疱疹病毒2型药物敏感性测定及其阿昔洛韦耐药株的建立

Method for Measuring Antiviral Susceptibility of Herpes Simplex Virus Type 2 and Establishment of Resistant Strain

目的应用空斑抑制试验测定单纯疱疹病毒2型(HSV-2)药物敏感性,并建立体外耐药病毒株。方法将HSV-2接种于乳兔肾细胞中,加入不同浓度的阿昔洛韦(ACV),培养72h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-2的药物敏感性。将HSV-2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-2标准株Sav毒株的IC50为1.1μg/mL。HSV-2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8.4μg/mL,56.9μg/mL和121.3μg/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-2的耐药性研究。

Objective To develop the method for measuring antiviral susceptibility and to establish an acyclovir（ACV） resistant herpes simplex virus type 2 （HSV-2）. Methods Baby hamster kidney cells were inoculated with HSV-2, and ACV was added to the medium in various concentrations. After 72 hours incubation, plaques were counted and the 50% inhibiting concentration（IC50 ）was calculated. HSV-2 was incubated in the presence of ACV for 9 passages. The IC50 of ACV was determined after the 3rd, 6th and 9th passage. Results The IC50 of ACV to HSV-2 Say was 1. 1μg/mL. In the presence of ACV, emergence of drug resistance was observed after passage 3 ; the IC50 of ACV after the 3rd, 6th and 9th passage was 8.41μg/mL,56. 9μg/mL and 121.3μg/mL respectively. Conclusion Plaque reduction assay is an efficient method for determination of antiviral drug sus- ceptibility of HSV and the establishment of ACV-resistant strain is essential for the study of resistance.