[Identification and sequence analysis of E gene of Dengue virus type 2 strain isolated from patient serum in Shenzhen].
OBJECTIVE
To isolate and identify the pathogen of Dengue fever from Shenzhen city in 2005 - 2006, and to analyze the molecular characteristics of the isolated Dengue virus strain as well as to explore its possible origin.


METHODS
IgM and IgG of serum samples taken from 60 suspected Dengue fever patients were detected by ELISA and immunochromatography, and 9 specimens were positive. Nine samples from patients with early stage Dengue fever were used to isolate virus with C6/36 cell line and the positive cell cultures were identified by MGB fluorescent PCR. The type of isolated virus strain was determined by RT-semi-nested-PCR and fluorescent PCR. E gene of isolated virus strain was amplified by RT-PCR and sequenced. Homology and phylogenetic tree of E gene of Shenzhen Dengue virus with the strains isolated from other areas were constructed.


RESULTS
Of nine antibody-positive serum samples, one strain of Dengue virus was successfully isolated. The isolated virus strain was confirmed as Dengue virus type 2 and designated as DEN2-SZ0521. The homology of nucleotide sequence and the deduced amino acid sequence of E gene of SZ0521 with standard type 2 Dengue virus NGC strain was 94.2% and 98.2%, but the homology with standard Dengue virus 1, 3, 4 in the same fragment were 59.1%, 57.2%, 58.5% and 68.1%, 66.7%, 63.2%, respectively. The phylogenetic tree indicated that SZ0521 had the greatest similarity with the Malay0412a/Tw strain and they lied in the same branch of the phylogenetic tree. The corresponding homology of nucleotide sequence and amino acid sequence was 99.8% and 100%, respectively. The isolated Dengue virus type 2 belonged to genotype IV with Indonesia-76, Somalia-84 and Sri Lanka-90.


CONCLUSIONS
Dengue virus was isolated from Shenzhen for the first time, and it was classified as type 2. It was confirmed that the type 2 Dengue virus may come from the epidemic area in Malaysia.
Yang
Fan
F
Shenzhen Center for Disease Prevention and Control, China. sailing_728@163.com
He
Jian-fan
JF
Xian
Hui-xia
HX
Zhang
Hai-long
HL
He
Ya-qing
YQ
Yang
Hong
H
Yao
Xiang-jie
XJ
D000330:Aedes / Q000821:virology; D000818:Animals; D002681:China; D003715:Dengue / Q000821:virology*; D003716:Dengue Virus / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification*; D005814:Genes, Viral; D006801:Humans; D010802:Phylogeny; D020539:Sequence Analysis, Protein; D017423:Sequence Analysis, RNA