[Expression of recombinant VP2 gene in insect sf9 cells and screening of clinical specimens].
OBJECTIVE
To clone and express VP, gene from HBoV, and the expressed VP, protein was as the antigen in order to detect serum from children in Wenling area with lower respiratory tract infections.


METHODS
The VP, gene was recombined with the genome of Baculovirus, which infected the insect cell. The fusion protein with HA tag was applied to confirm the specificity of expressed protein. Furthermore, the recombinant protein was observed using electron microscopy. The 176 serum from children in Wenling area with lower respiratory tract infections was screened using Western blot.


RESULTS
The expressed VP2 protein was more than 60% in total proteins from insect cell, and MWt about 60 x 10(3). The virus-like particle (VLP) was observed using electron microscopy, and size about 20 nm. The 176 serum from children in wenling area with lower respiratory tract infections was screened using Western blot. The HBoV positive rate was 2.28% (4/176).


CONCLUSIONS
The VP2 protein from human bocavirus was expressed in insect cell successfully. Through HA tag the VP2 protein was specific, and then the assay using SDS-PAGE with Western blot could detect and screen the antibody in serum from children with lower respiratory tract infections rapidly and accurately.
Teng
Ling-fang
LF
Wenling Afiliated Hospital, Wenzhou Medical College, Zhejiang 317500, China.
Lin
Feng
F
Zheng
Me-yun
MY
Zheng
Chang-hua
CH
Wu
Feng
F
Zeng
Ai-ping
AP
Huang
En-pei
EP
Mo
Yi-han
YH
Zheng
Min-qiao
MQ
Li
Xu-yang
XY
Hou
Jian-yi
JY
D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D052660:Bocavirus / Q000235:genetics* / Q000276:immunology; D036022:Capsid Proteins / Q000235:genetics* / Q000276:immunology; D002675:Child, Preschool; D005260:Female; D015870:Gene Expression; D006801:Humans; D007223:Infant; D008297:Male; D010322:Parvoviridae Infections / Q000097:blood / Q000175:diagnosis* / Q000276:immunology / Q000821:virology; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D018411:Spodoptera
D000914:Antibodies, Viral; D036022:Capsid Proteins; D011994:Recombinant Proteins