First isolation and identification of Zika virus in China
10.3760/cma.j.issn.0254-5101.2016.04.002
- VernacularTitle:我国首例寨卡病毒的分离与鉴定
- Author:
De WU
;
Qiqi TAN
;
Jiufeng SUN
;
Huiqiong ZHOU
;
Dawei GUAN
;
Huan ZHANG
;
Dan NING
;
Changwen KE
- Publication Type:Journal Article
- Keywords:
Zika virus;
Viral isolation;
Arbovirus;
Suckling mice
- From:
Chinese Journal of Microbiology and Immunology
2016;36(4):247-251
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish a method for the isolation of Zika virus and to gather experi-ences for viral isolation. Methods Suckling mice at age 1-3 days were inoculated with serum samples posi-tive for Zika virus through intracranial injection. All mice were sacrificed 6 days after the injection. Viral nu-cleic acids were extracted from brain, heart, liver, spleen, lung, kidney, muscle, skin and intestine tissue samples and analyzed by real-time RT-PCR. The supernatants of brain tissues positive for Zika virus were used for subculturing. Nested PCR was performed to amplify the NS5 gene of the isolated virus. The se-quences of NS5 gene were analyzed by using MEGA6. 0 software. Results All of the tissue samples were positive for Zika virus. Higher viral loads were detected in heart and brain tissue samples with cycle thresh-old (Ct) values of 24. 4 and 25. 3, respectively. The second generation of Zika virus was identified in suck-ling mice brain tissues 2 days after infection by using real-time RT-PCR. The amplified product of nested PCR was 972 bp in length. Sequencing analysis showed that the isolated Zika virus ( GDZ16002 strain) be-longed to the Asian lineage. Conclusion A strain of Zika virus was successfully isolated in China by using intracranial injection via a suckling mouse model. The isolated Zika virus belonged to the Asian lineage.
