[Isolation and cell culture of human bocavirus (HBoV) by human bronchial epithelial cell lines].
OBJECTIVE
To investigate pave a way for studying pathogenicty of HBoV.


METHODS
Isolation and cell culture of HBoV by human bronchial epithelial cell line, which was founded in our laboratory. The morphology of the virus were primarily studied with a transmission electron microscope. In addition, transcript mRNA was detected in human bronchial epithelial cells, which was passaged and infected within HBoV, using the reverse-transcription polymerase chain reaction (RT-PCR). The amplified products nucleotide sequence of HBoV were sequencing and sequence analysis.


RESULTS
Cytopathic effect (CPE) was observed after the aseptic residue of filtration of 2 case sputum specimens with HBoV, which was inoculated to the human bronchial epithelial cell line. The virus particles were observed in the cytoplasm, which were hexagonal or spherical in shape and 18-26 nm in diameter,bulk was 20 nm. cDNA amplicon obtained 295 bp fragment results of electrophoresis bands as same as NS1 region of the conserved matrix gene of publish sequence of HboV. PCR products nucleotide sequence of HboV were compared with corresponding HboV GeneBank sequences. The comparison/alignment and construction of phylogenetic trees also point to an affiliation of the parvovirus to the species HBoV.


CONCLUSIONS
Isolation and identification of HBoV could be done in the human bronchial epithelial cell, and we found some characterizing CPE in the human bronchial epithelial cell after HBoV infection. The above studies pave a way for studying pathogenicty of human bocavirus.
Lin
Feng
F
Wenling Afiliated Hospital, Wenzhou Medical College, Zhejiang 317500, China.
Teng
Ling-fang
LF
Zheng
Mei-yun
MY
Zheng
Chang-hua
CH
Wu
Feng
F
Li
Hua
H
Zheng
Ming-qiao
MQ
Zeng
Ai-ping
AP
Hung
En-pei
EP
Mo
Yi-han
YH
Hou
Jian-yi
JY
D001980:Bronchi / Q000166:cytology* / Q000821:virology; D018929:Cell Culture Techniques; D002460:Cell Line; D002648:Child; D002675:Child, Preschool; D004847:Epithelial Cells / Q000821:virology*; D056661:Human bocavirus / Q000145:classification / Q000235:genetics / Q000254:growth & development* / Q000302:isolation & purification*; D006801:Humans; D007223:Infant; D008297:Male; D008969:Molecular Sequence Data; D010322:Parvoviridae Infections / Q000821:virology*; D010802:Phylogeny; D012141:Respiratory Tract Infections / Q000821:virology*; D014776:Virus Cultivation