1.A new member of Brevidensovirus, 0507JS11 virus isolated from Culex mosquitoes collected in Xinjiang.
Xin-jun LÜ ; You-gang ZHAI ; Xiao-hong SUN ; Shi-hong FU ; Huan-qin WANG ; Su-xiang TONG ; Song ZHANG ; Guo-dong LIANG
Chinese Journal of Preventive Medicine 2009;43(5):385-389
OBJECTIVETo probe the primary characteristic of 0507JS11 virus isolated from Culex sp. and determine the classification of 0507JS11 virus in taxonomy.
METHODS0507JS11 virus was cultured in Aedes albopictus C6/36 cells and cytopathic effects (CPEs) were recorded. Electro-microscopic morphology of 0507JS11 virus was observed. Total DNA extract of 0507JS11 virus was detected by 1% Agarose Gel Electrophoresis. Complete genomic sequence of 0507JS11 virus was sequenced and then made phylogenetic analysis.
RESULTS0507JS11 virus could cause CPEs in Aedes albopictus C6/36 cells. Viral particles have no envelope and appear icosahedron symmetry with diameter of 20 nm. The genome of 0507JS11 virus was positive single strand DNA (ssDNA) with full length of 3977 nt. However, a DNA band about 4 kbp was observed in the electrophoresis of total DNA extract of 0507JS11 virus. The coding region of the genome included three ORFs, ORF1 and ORF2 code NSP1 and NSP2, ORF3 codes VP. Phylogenetic analysis of the complete genomic sequence of 0507JS11 virus indicated an independent linear in Brevidensovirus.
CONCLUSION0507JS11 virus is a new member in Brevidensovirus.
Animals ; Culex ; virology ; DNA, Viral ; genetics ; Densovirinae ; classification ; genetics ; isolation & purification ; Genome, Viral ; Sequence Analysis, DNA
3.Sequence analysis of the complete genome of bocavirus WLL-1.
Feng LIN ; Ai-Ping ZENG ; En YANG ; Hai-Yan LIN ; Chang-Hua ZHENG ; Hong CHEN ; Huai LI ; Xu-Yang LI ; Ming-Sul YU ; Ning-Min YANG ; Da-Zhi JIN ; Guang-Chuang YU ; Xiao-Chen BO ; Si-Yuan WEN ; Sheng-Qi WANG
Chinese Journal of Virology 2007;23(1):57-59
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.
Bocavirus
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classification
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genetics
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isolation & purification
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China
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DNA, Viral
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chemistry
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genetics
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Genome, Viral
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Humans
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Molecular Sequence Data
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Phylogeny
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Sequence Analysis, DNA
4.Surveillance of viral contamination of invasive medical instruments in dentistry.
Lin-fu ZHOU ; Hai-hong ZHU ; Jun LIN ; Min-jun HU ; Feng CHEN ; Zhi CHEN
Journal of Zhejiang University. Science. B 2006;7(9):745-748
OBJECTIVETo investigate the viral contamination of invasive medical instruments in dentistry and to provide health administrative institutions with surveillance data.
METHODSSterilized samples were randomly collected from the department of dentistry to detect HBV-DNA, HCV-RNA, HIV-RNA and HBsAg.
RESULTSOf the invasive medical instruments that were sterilized with 2% glutaraldehyde, one of the samples was positive for HBV-DNA, and another sample was positive for HBsAg.
CONCLUSIONThough massive virus contamination of invasive medical instruments in dentistry has been reduced to a low level, the occurrence of contamination still remains.
DNA, Viral ; analysis ; Dental Instruments ; virology ; Equipment Contamination ; HIV ; isolation & purification ; Hepacivirus ; isolation & purification ; Hepatitis B Surface Antigens ; analysis ; Hepatitis B virus ; isolation & purification ; Humans ; RNA, Viral ; analysis
5.Analysis on the SARS-CoV genome of PUMC01 isolate.
Ke ZOU ; Hua ZHU ; Ke-yue DING ; Zhong WANG ; Yong LIU ; Ting WANG ; Jian YANG ; Guo-zhu WEI ; Xin-feng ZHOU ; Wen ZHANG ; Zhan-xia YU ; Zheng FAN ; Xiao-zhong PENG ; Chuan QIN ; Xiang-jun LIU ; Yan SHEN ; An-ping NI ; Bo-qin QIANG
Acta Academiae Medicinae Sinicae 2003;25(5):495-498
OBJECTIVETo perform variation and phylogenetics analysis on the SARS-CoV genome sequence (PUMC01) isolated in the Peking Union Medical College Hospital.
METHODSThe cDNA library of SARS-CoV (PUMC01 isolate) was constructed by means of random-priming strategy. Random selected plasmid was sequenced and the genome sequence of SARS-CoV-PUMC01 was assembled by conventional methods (The Genebank Accession No. of SARS-CoV-PUMC01 is AY350750). The variation and phylogenetics analysis were performed by comparing the PUMC01 sequence with other SARS-CoV isolates.
RESULTSTen variation sites were found by comparing PUMC01 isolate with Tor2 and Urbani isolates. In phylogenetic analysis of 18 SARS-CoV isolates, two classes were observed and there is different differential time between these two classes and the different isolates in each class.
CONCLUSIONSThe evidence of phylogenetic analysis of different SARS-CoV isolates from different region is instructive for understanding the clinical relations between the different isolates and the transmission chain of SARS-CoV.
Amino Acid Sequence ; Base Sequence ; China ; DNA, Viral ; genetics ; Genetic Variation ; Genome, Viral ; Molecular Sequence Data ; Phylogeny ; SARS Virus ; genetics ; isolation & purification ; Sequence Analysis, DNA ; Viral Proteins ; genetics
6.Detection of Epstein-Barr virus DNA in plasma/serum: a useful serological indicator for diagnosis of nasopharyngeal carcinoma.
Shijuan MAI ; Yongsheng ZONG ; Min ZHANG ; Biling ZHONG ; Suxia LIN
Chinese Medical Journal 2002;115(12):1895-1897
OBJECTIVETo compare the detection rates of Epstein-Barr virus (EBV) DNA in the serum/plasma between apparently healthy adults (AHAs) and nasopharyngeal carcinoma (NPC) patients in attempt to evaluate the efficiency of EBV DNA assay for serodiagnosis of NPC.
METHODSThe plasma and serum were obtained from 58 AHAs and 66 untreated NPC patients. EBV DNA W-fragment was detected using nested ploymerase chain reaction (PCR). Immunoenzymatic assay for titration of IgA-VCA was also adopted.
RESULTSEBV DNA detection rate (84.85%) in the plasma/serum of 66 NPC patients was significantly higher than that (10.34%) in 58 AHAs. The sensitivity of plasma/serum EBV DNA assay (0.8485) was higher than that (0.8030) of titrating IgA-VCA (positive criterion >/= 1:40) though the specificities of these two tests were the same (0.8966). The correct rate, predictive value of a positive test, and Odds ratio of dual positivity (0.8387, 0.9792 and 141.0, respectively) were higher than those of single positivity either to plasma/serum EBV assay (0.5242, 0.7333 and 1.1423, respectively) or to IgA-VCA >/= 1:40 test (0.4839, 0.5385 and 1.0480, respectively).
CONCLUSIONThe EBV DNA detection in the plasma/serum using nested PCR may be a useful indicator for serodiagnosis of NPC.
Antigens, Viral ; blood ; DNA, Viral ; blood ; Herpesvirus 4, Human ; isolation & purification ; Humans ; Immunoglobulin A ; blood ; Nasopharyngeal Neoplasms ; diagnosis ; virology ; Serologic Tests
7.Absence of Human Papillomavirus DNA in Nongenital Seborrheic Keratosis.
Eun So LEE ; Mi Ran WHANG ; Won Hyoung KANG
Journal of Korean Medical Science 2001;16(5):619-622
Seborrheic keratosis (SK) is a benign epidermal tumor of unknown etiology. Because of its wart-like morphology, human papillomavirus (HPV) has been suggested as a possible causative agent. Viral involvement, however, has not been confirmed yet despite extensive research. The aim of this study was to evaluate the presence of HPV 6/11, 31, 33 DNA in nongenital SK. We analyzed 40 biopsy specimens taken from patients with nongenital SK using in situ polymerase chain reaction (PCR) and PCR with tissue extracts. The SK specimens (n=40), analyzed by in situ PCR, were negative for all HPV probes tested (types 6/11, 31, 33). Control slides (condyloma acuminatum, n=3) were positive for type 6/11, 31, and 33 HPV probes tested. Melasma samples (n=4), the negative controls, were consistently negative. No HPV DNA band was detected by PCR with the tissue extracts from paraffin-embedded SK samples, while condyloma acuminatum, the positive controls, showed DNA bands of the correct molecular weights. Our results show that HPV type 6/11, 31, and 33 cannot be recognized as causative agents for nongenital SK, which is in contrast to the previous studies. Further studies are required to reveal the presence of other types (more than 90) of HPV DNA.
DNA, Viral/*analysis
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Female
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Human
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Keratosis, Seborrheic/*virology
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Male
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Papillomavirus, Human/*isolation & purification
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Polymerase Chain Reaction
8.Detection of food-borne rotavirus by molecular motor biosensor.
Jie ZHANG ; Meiling XU ; Xuan WANG ; Yu WANG ; Xiaojin WANG ; Yan LIU ; Dezhou GU ; Guangquan CHEN ; Peirong WANG ; Jiachang YUE
Chinese Journal of Biotechnology 2013;29(5):681-690
To develop a specific, rapid and convenient method based on molecular motor biosensor to detect food-borne rotavirus. A specific probe was encompassed the conservative region of rotavirus's VP7 segment, and a molecular motor detect device was constructed by connecting probes to F0F1-ATPase molecular motor through biotin-streptavidin system. This biosensor's sensitivity was 0.005 ng/mL for rotavirus RNA. Extracted virus RNA was conjugated with the biosensor separately, at the same time ATP was synthesized. By comparing fluorescence intensity, we can detect rotavirus RNA in samples. This method possessed specificity for rotavirus, without any cross-reaction with Hepatitis A virus and noroviris, and it could be accomplished within 1 h. We detected 15 samples using this method and the results were compared with RT-PCR results. This method is sensitive and specific for rotavirus, and it can be used to detect food-borne rotavirus.
Biosensing Techniques
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methods
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DNA, Viral
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analysis
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genetics
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Food Microbiology
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methods
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Rotavirus
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genetics
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isolation & purification
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Sensitivity and Specificity
9.Replication of duck plague virus in artificially infected ducks detected by in situ hybridization.
An-Chun CHENG ; Yong-Hong LIAO ; De-Kang ZHU ; Ming-Shu WANG ; Gui-Ping YUAN ; Chao XU ; Xiao-Ying HAN
Chinese Journal of Virology 2008;24(1):72-75
Replication of duck plague virus(DPV) in artificially infected ducks were detected by in situ hybridization (ISH) which employed a 37bp oligonucleotide as probe designed according to DPV DNA sequence in GenBank. The results indicated that DPV DNA was detected in liver, intestine and bursa Fabricius at 4 h, in spleen and esophagus at 6h, in thymus at 12h post infection; DPV DNA in lung and kidney was detected only in dead ducks and no positive signal was detected in muscle, heart, cerebrum and pancreas. DPV DNA was distributed in cell nucleus and cytoplasm. Hepatocytes, sinus endodermal cells and Kuffer's cells were the mainly infected cell types in liver. DPV DNA was mainly detected in epithelium of villi, in lamina propria of intestinal villi of duodenum, in stratum spinosum of esophagus, and in epithelium, cortex, medulla of bursa Fabricius. The positive signals were mainly detected in medulla of thymus, lymphocytes and macrophages of spleen. The research suggests that ISH is a direct and specific method in detecting DPV DNA in paraffin sections and it's also a good method for virus diagnosis and DNA location of DPV.
Animals
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DNA, Viral
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analysis
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Ducks
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virology
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In Situ Hybridization
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Influenza A virus
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genetics
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isolation & purification
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physiology
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Virus Replication
10.Characterization of a novel podoviridae-phage infecting Serratia marcescens isolated in China.
Feng-Yu XU ; Yong-Jie LIU ; Hong-Xia MA ; Yan ZHANG ; Sheng-Bing SU ; Chan-Juan SHEN ; Cheng-Ping LU
Chinese Journal of Virology 2012;28(4):439-443
Serratia marcescens jn01 was employed as the host for the isolation of phages from environmental sewage. One strain of phage named SmPjn was purified by picking transparent plaque with 2mm diameter and clear edge on the double-layer agar repeatedly. Electron micrographs indicated that the phage head was icosahedral with head size and tail length of (58 +/- 2.16) x (55 +/- 0.47) nm and (7 +/- 1.25) nm, respectively. On the basis of the morphology, this phage belongs to the family Podoviridae. Host-range determination revealed that the phage was capable of infecting the other two isolates of S. marcescens, P25 and CMCC41002. The optimal multiplicity of infection was 1. A one-step growth curve of SmPjn indicated that the latent period and burst size were estimated at 50 min and 1,125 pfu/cell, respectively . Genomic DNA of SmPjn was above 27kb in size and could be digested by Hind Ill and EcoR I into 11 and 9 visible fragments after electrophoresis, respectively. A novel Podoviridae-phage infecting S. marcescens was firstly reported in China.
China
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DNA, Viral
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genetics
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isolation & purification
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metabolism
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Host Specificity
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Podoviridae
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genetics
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growth & development
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isolation & purification
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Restriction Mapping
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Serratia marcescens
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physiology