Preparation and identification of influenza H1N1 subtype vaccine candidate strain in China
10.3760/cma.j.issn.1003-9279.2017.04.016
- VernacularTitle: 2015—2016年度中国甲型H1N1亚型流感病毒疫苗候选株的制备及鉴定
- Author:
Jing TANG
1
,
2
;
Li XIN
;
Xiaodan LI
;
Yongkun CHEN
;
Junfeng GUO
;
Weijuan HUANG
;
Yanhui CHENG
;
Dayan WANG
;
Yuelong SHU
Author Information
1. National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention
2. Key Laboratory for Medical Virology, Ministry of Health, Beijing 102206, China
- Publication Type:Journal Article
- Keywords:
Influenza A virus;
Classical reassortment;
H1N1 subtype;
Chinese epidemic strain
- From:
Chinese Journal of Experimental and Clinical Virology
2017;31(4):348-352
- CountryChina
- Language:Chinese
-
Abstract:
Objective:Influenza H1N1 subtype vaccine candidate strains from a 2015—2016 year epidemic strain in China were prepared and identified by themethod of classical reassortment.
Methods:The influenza H1N1 epidemic strain and H3N2 high-yield reassortant parental strain (X-157) were mixed and inoculated into embryonated chicken eggs by the classical reassortmentmethod . The negative selection of mixed culture virus was carried out with the antiserum of H3 protein and the antiserum of X-157 strain. Real-time PCRmethod was used to test the HA and NA genes. Restriction enzyme digestionmethod was used to identify the internal genes. HA and NA genes of selected strains were sequenced. The strain which HA and NA genes possessed the same amino acid constitution with the wild type virus was selected and immunized to ferret. Two-way test was carried out.
Results:Five strains with expected HA and NA genes were selected by real-time PCR. Internal genes were identified, with 4 strains had 6+ 2 constitution, 1 strain had 5+ 3 constitution. Comparing with the wild type virus, HA and NA genes of the 5 strains had no mutation. HA titer of reassortant strains was above 1 024. HI titer of the selected NO.12 reassortment strain reached 5 120, and two-way test was passed. The yield of reassortant strain was 64 times that of the wild type strain.
Conclusions:A circulating influenza A (H1N1) strain of influenza A (2015—2016) was successfully prepared in China and laid the foundation for vaccine storage and disease prevention and control.
