ParvoDB ID PV027024 GenBank Accession GQ403973
Subfamily Parvovirinae Genus Bocaparvovirus
Species Bocaparvovirus primate1 Sequence Length 2016
Definition Human bocavirus isolate R3071126003 capsid protein (VP1) gene, complete cds. Submission Date 2009-07-21
Strain Name R3071126003 Sampling Date 2007-11-26
Sampling Country China Location the Shanghai Nanxiang Hospital, China
Submitted By Wang,K., Wang,W., Shen,J., Yan,H., Ren,P., Zhang,J. and Deubel,V. Submitting Institution Unit of Emerging Viruses, Institut Pasteur of Shanghai, Chinese Academy of Sciences, Shanghai Institute of Biological Sciences, 411 Hefei Road, Shanghai 200025, P.R. China
Original Host Homo sapiens Sample Type nasal swabs, throat swabs, blood, as well as urine samples
Standardized Host Name Homo sapiens Host Tax Rank species
Standardized Host Class Mammalia Standardized Host Order Primates
Environmental Origin - N/A - Reagent Origin - N/A -
Number Reagent/Consumables Purpose PMID/Url Reference
1 Viral Transport Medium Storage medium for collected nasal and throat swab samples. 20022295 View Related Publication
2 3.2% Sodium Citrate Anticoagulant in blood sample collection tubes.
3 QIAamp Viral RNA Mini Kit (QIAGEN, Hilden, Germany) Extraction of total RNA and DNA from specimens.
4 RNase-free water Elution of nucleic acids.
5 Takara Premix Ex TaqTM Perfect Real Time (Takara Biotechnology, Dalian, China) Real-time PCR assays for the detection of HBoV.
6 Primers and Probe (targeted to the NP1 gene) Used in real-time PCR assays to detect HBoV.
7 ABI PRISM 7900HT (Applied Biosystems, Foster City, CA, USA) Equipment for running and analyzing real-time PCR assays.
8 pskHBoV containing the whole genome of HBoV ST2 genotype Used as a standard for generating the standard curve in PCR.
9 One-Step RT-PCR kit (QIAGEN) Conventional RT-PCR to detect HBoV and other respiratory viruses.
10 Multiplex RT-PCR reagent (described previously) Diagnosis of other respiratory viruses in HBoV-positive samples.
11 QIAGEN’s One-Step RT-PCR kit (QIAGEN) Amplification of the entire VP1 gene for sequencing.
12 QIAquick Gel Extraction Kit (QIAGEN) Extraction of PCR product fragments for sequencing analyses.
13 pMD18T (Takara Biotechnology) Cloning vector for sequencing.
14 Biosune Sequence Company and Life Biotechnology (Shanghai, China) Services for sequencing the amplified products.
15 MEGA version 3.1 software Software for phylogenetic analysis (excluded as it's software).
16 Spodoptera frugiperda Sf21 cells Host cells for infection to recover recombinant baculovirus.
17 High Five cells Cells infected with recombinant baculovirus to express VP2 proteins.
18 NaHCO3 (25 mM) Solution for cell lysis.
19 Ammonium sulfate (20%) Precipitation of proteins.
20 PBS (phosphate-buffered saline) Resuspending proteins and dialysis.
21 pET30a (expression vector containing 6xHis tag) Cloning and expression of VP2 fragments in E. coli.
22 E. coli BL21(DE3) host strains Host strains for expression of recombinant proteins.
23 IPTG Induction of protein expression in E. coli.
24 Ni-NTA agarose (Invitrogen) Purification of His-tagged recombinant proteins.
25 SDS-PAGE gel (12.5%) Separation of proteins for immunoblotting.
26 Nitrocellulose membranes Transfer of proteins for Western blotting.
27 TBS-T (0.5% Tween-20 in TBS buffer) Blocking and washing buffer during immunoblotting.
28 Nonfat dry milk (5%) Blocking solution in immunoblotting.
29 Patient serum samples Primary antibodies for detection in immunoblotting.
30 Anti-human IgG or IgM conjugated with horseradish peroxidase (HRP) (Bethy Laboratories, Montgomery, TX, USA) Secondary antibodies for detection in immunoblotting.
31 3,3′-diaminobenzidine and H2O2 Substrates to reveal peroxidase activity in Western blot.
32 SARS-HCoV spike protein produced in E. coli Negative control antigen for ELISA.
33 o-Phenylenediamine Dihydrochloride (OPD) (Sigma, St. Louis, MO, USA) Substrate for HRP in ELISA.