Detection of BK virus infection in renal transplant recipients and clinical application / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To study the detection methods of BK virus infection in kidney transplant recipients, and to explore the clinical application.</p><p><b>METHODS</b>132 cases of renal transplant recipients were undertaken BK virus detection including presence of decoy cells in urinary sediment, urine and serum BKV-DNA to demonstrate the BK virus replication.</p><p><b>RESULT</b>Among 132 cases of renal transplant recipients, urinary decoy cell was found in 37 (28.0%) patients and the median time was 12 months after surgery. 32 (24.2%) patients were diagnosed as BK viruria at a median of 11 months after surgery, and 16 (12.1%) recipients were diagnosed as BK viremia at a median of 15 months after surgery, 5 patients with BK viruria were diagnosed as BK virus associated nephropathy according to allograft biopsy.</p><p><b>CONCLUSION</b>To make early diagnosis of BK virus infection, detection of urine decoy cells and BKV-DNA in urine and plasma sample is important,which provides an important basis for the prevention of BK virus associated nephropathy.</p>

Clinical features and gene mutation analysis in Machado-Joseph disease of spinocerebellar ataxia type 3 in littoral of Zhejiang / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To study the clinical features and gene mutation analysis in Machado-Joseph disease of spinocerebellar ataxia type 3 in littoral of Zhejiang.</p><p><b>METHODS</b>Clinical manifestation and brain MRI data 18 patients with SCA in family were analyzed. The gene mutations of 18 patients and 10 family numbers without abnormal presentation, and 12 healthy persons of controls.</p><p><b>RESULTS</b>The gene mutations of 18 patients is SCA3/MJD, and 2 asymptomatic SCA3/MJD had been detected in SCA family. Normal alleles of SCA3/MJD have CAG repeats ranging from 14 to 27, patients from 67 to 82, asymptomatic and carrier SCA3/MJD from 28 to 45. The main features of 18 patients included gait ataxia, ambiguity in speech and action clumsiness. Brain MRI showed remarkable atrophy on cerebellum and brain stem.</p><p><b>CONCLUSION</b>CAG expansions were related to SCA3/MJD. The clinical manifestations are ataxia and dysarthria. The detection of repeated times CAG can provide an effective way for the genetic and asymptomatic diagnosis.</p>

Expression of recombinant VP2 gene in insect sf9 cells and screening of clinical specimens / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>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.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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).</p><p><b>CONCLUSION</b>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.</p>

Isolation and cell culture of human bocavirus (HBoV) by human bronchial epithelial cell lines / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To investigate pave a way for studying pathogenicty of HBoV.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSION</b>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.</p>

Discovery and identification of WU polyomavirus in children from Zhejing region / 病毒学报

WU polyomavirus, which was firstly discovered in 2007, is a new human polyomavirus belonging to Polyomaviridae and containing circular double-stranded genomic DNA. In this study, the 278 clinical sputum specimens from children under 5 years old were collected from Wenzhou Medical College affiliated Wenling First Hospital, Zhejiang Province. Based on identification assay of WU polyomavirus previously reported, a WU polyomavirus was identified from clinical samples successfully, the positive rate was 0.4%. The sequences of PCR products were identical to that of VP2 gene and large T antigen gene derived from WU polyomavirus reported. The above results strongly suggested that the WU polyomavirus isolated was firstly found in Chinese children with acute lower respiratory tract infections. This study provides a firm basis for further research of WU polyomavirus.

Characterization of the cytopathic effect in human bronchial epithelial cell after Human Bocavirus Infection (HBoV) / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>In this study, human bronchial epithelial cells were inoculated with positive sputum specimens of HBoV. After four days' infection, cytopathic effects (CPE) were observed by inverted microscopy. These viruses all cause typical cell damages such as rounded and shrivelled, fusion and fallout. These damages got quick following increased future degenerations. The other assay result of CPE within the infected cells were observed by inverted microscopy, have typical "owl's eye" plaque and above 90 percent hemadsorption within the infected cells by erythrocytes for hemadsorption technique. The typical fluorescence lump of nucleus within the infected cells was found by indirect immunofluorescence technique.</p><p><b>CONCLUSION</b>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 pathogenicity of human bocavirus.</p>

Clinical application of microarray technique to quantify the lamivudine-resistant gene mutant of hepatitis B virus / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To evaluate clinical applicability of a novel technique that can quantify the lamivudine-resistant mutants of hepatitis B virus (HBV) in the serum of patients utilizing gene microarray technology.</p><p><b>METHODS</b>The oligonucleotide microarray was designed to detect 3 important mutational positions. Fifty-one patients who were receiving lamivudine therapy were selected as subjects. The oligonucleotide microarray and traditional sequencing were applied to detect the lamivudine resistant mutation, the monitoring lasted for 24 months. Then the clinical result was analyzed and the obtained data were compared between the two methods.</p><p><b>RESULTS</b>Lamivudine resistant mutation was detected in 39 percent of the patients during the 2 years period. The results of the oligonucleotide microarray technique was consistent to the results of traditional sequencing in accuracy and the miroarray was more sensitive in detection of the mixed infection.</p><p><b>CONCLUSION</b>Application of the oligonucleotide microarray for quantitative detection of lamivudine-resistant mutation of HBV is feasible.</p>

Clinical prospective study on maternal-fetal transmission of human bocavirus / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To investigate maternal-fetal transmission at human bocavirus (HBoV).</p><p><b>METHODS</b>IgG antibody to HBoV in serum samples of 316 mothers were determined with ELISA and HBoV DNA was determined with real time PCR in the sera of the mothers and their infants.</p><p><b>RESULTS</b>HBoV-IgG was positive in 40.20 percent (127/316) of the mothers, while it was positive in 29.43 percent (93/316) of the cord blood specimens of the infants. The difference between the two groups was significant (X2=8.12, P less than 0.005); 93 samples of both the mothers and the infants were positive for HBoV-IgG.</p><p><b>CONCLUSION</b>HBoV-IgG can cross the placenta to the fetuses through placenta. Further study is needed to answer the question whether vertical maternal-fetal transmission occurs.</p>

Sequence analysis of the complete genome of bocavirus WLL-1 / 病毒学报

Human bocavirus, which was firstly discovered in 2005, is a new human parvovirus associated with lower respiratory tract infection in children. In this study, a human bocavirus, named WLL-1 isolate, was identified in Wenlin County, Zhejiang Province. The genome of bocavirus WLL-1 has been sequenced and analyzed. Phylogenetic analyses showed that WLL-1 shares 99% homology with other bocaviruses recently reported, but also has some special variations.