Practical applications of different inoculation methods in the isolation of herpes simplex virus
10.3760/cma.j.cn112309-20241022-00358
- VernacularTitle:不同接种方法在单纯疱疹病毒分离中的实践应用
- Author:
Hui WANG
1
;
Jieqiong ZHANG
1
;
Ying LI
1
;
Jiao WANG
1
;
Mingming WANG
1
;
Haotian LI
1
;
Dongbo YU
1
;
Shiwen WANG
1
;
Xuexin LU
1
Author Information
1. 中国疾病预防控制中心病毒病预防控制所 传染病溯源预警与智能决策全国重点实验室，北京 102206
- Publication Type:Journal Article
- Keywords:
Herpes simplex virus;
Viral isolation;
Suspended method;
Mixed method
- From:
Chinese Journal of Microbiology and Immunology
2025;45(4):304-309
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To establish a protocol for virus isolation using the mixed method, and evaluate the efficacy of the suspended method and the mixed method in isolating herpes simplex virus (HSV).Methods:Simulated HSV-infected clinical samples were prepared using HSV-1 F strain and CDC-P1 strain. Both the suspended method and the mixed method were used to isolate HSV-1 from these samples. The virus isolation efficiency of the mixed method under various conditions was assessed. These conditions included different multiplicity of infection (MOI), cell seeding densities, and virus adsorption times.The 50% tissue culture infective dose (TCID 50) assay was used for the assessment. The positive rates of virus detection under low viral load conditions were compared between the two methods. Results:Under the conditions of a MOI of 0.005, a virus adsorption time of 15 min, and a cell seeding density of 1×10 6 cell/ml, the mixed method achieved effective isolation of HSV-1. When the virus titer of the sample was 100 TCID 50/ml, the positivity rate of the mixed method reached 100.0%, while the positivity rates of the suspended method were 50.7% (38/75) and 52.0% (39/75) after cultured for 72 h and 96 h, respectively. When the virus titer of the sample was 10 TCID 50/ml, the positivity rate of the mixed method was 100.0%, while the positivity rate of the suspension method was 0. Conclusions:The mixed method exhibits significantly higher efficiency in HSV isolation compared with the suspended method. Under the conditions of high viral load, both the suspended method and the mixed method can be effective in isolating HSV-1. For clinical samples with low viral loads, the mixed method has greater applicability.
