A multiplex liquid-chip assay based on Luminex xMAP technology for simultaneous detection of six common respiratory viruses.

We utilized one-step multiplex reverse transcription-PCR (RT-PCR) and Luminex xMAP technology to develop a respiratory multiplex liquid-chip assay (rMLA) for simultaneous detection of 6 common respiratory viruses, including influenza virus type A (FluA) and type B (FluB), para-influenza virus type 3 (PIV-3), respiratory syncytial virus (RSV), human metapneumovirus (MPV) and a threatening virus to China, Middle East Respiratory Syndrome coronavirus (MERS-CoV). Performance of rMLA was evaluated by comparing with real-time RT-PCR. Detection data from clinical specimens showed that the rMLA had diagnostic sensitivities of 97.

[Tracing investigation and diagnosis of one transfusion-transmitted malaria case in Zhejiang Province].

Objective: To identify the sources of infection and the mode of transmission of a malaria case with unknown origin.

Methods: Clinical data of the case were collected and the epidemiological investigation was conducted. The blood samples of the patient and the suspected infection source (blood donor) were detected by microscopy, rapid diagnostic test strip (RDT) and nested PCR.

[Laboratory testing for a case of imported Plasmodium ovale infection in Zhejiang Province].

Blood sample obtained from a patient, which returned from Equatorial Guinea, with clinical diagnosis of Plasmodium infection was confirmed as imported P. ovale infection by etiology and molecular biological methods. 50 microl blood was obtained before taking anti-malarial drugs to make thin and thick blood smears, Giemsa stained, and observed by microscopy.

[Secretory expression of salivary ATP diphosphohydrolase (apyrase) from Aedes albopictus in Pichia pastoris].

The gene-coding mature apyrase protein from Aedes albopictus was amplified by RT-PCR and cloned in frame with the a-factor secretion signal peptide into Pichia pastoris secreting expression vector pGAPZalpha-A resulting in the pGAPZa-A-apyrase. After being linearized by Bln I restriction enzyme, the recombinant pGAPZalpha-A-apyrase was trans-formed into Pichia pastoris GS115 by electroporation. Recombinant strains pGAPZalpha-A-apyrase/GS115 were screened on YPDS plates containing Zeocin and identified by PCR.

[Pathogen identification and clinical diagnosis for one case infected with Babesia].

Objective: To identify the pathogen and make diagnosis on a case who was misdiagnosed as malaria.

Methods: Clinical and epidemiological information of the suspected case was collected. Blood samples during hospitalization were collected and examined microscopically.

[Temporal and spatial population dynamics of rabies virus isolates in China].

In order to study phylogeography, population dynamics and molecular evolution of rabies viruses (RABVs) isolates from China, especially spatio-temporal dynamics, the timescale of RABVs evolution and its pattern of migration, we performed an extensive comparative analysis of RABV N gene sequence data, representing 167 isolates sampled from 20 provinces in a 78-year period (from 1931 through 2009). The available Chinese isolates could be divided into two distinct clades:Phylogroup clades I comprised Chinese group 1-4; Phylogroup clades II contained Chinese group 5-8. We found no evidence for positive selection (dN/dS>1) acting at any codon and found strong selective constraints for N gene.

[Molecular epidemiology investigation of rabies virus nucleoprotein genes in China].

Objective: To study the relationship between the distribution of rabies virus and genetic variation, the genetic characterization and variation of rabies virus strains in China were analyzed.

Methods: The downstream 720 nucleotides of Nucleoprotein (N) gene coding region of the rabies specimens from different areas and host animals were sequenced, and then homology and phylogenesis were analyzed.

Results: Nucleotide similarities of 34 N gene sequences were 87.

[Sequencing and analysis of complete genome of rabies viruses isolated from Chinese Ferret-Badger and dog in Zhejiang province].

Based on sequencing the full-length genomes of four Chinese Ferret-Badger and dog, we analyze the properties of rabies viruses genetic variation in molecular level, get the information about rabies viruses prevalence and variation in Zhejiang, and enrich the genome database of rabies viruses street strains isolated from China. Rabies viruses in suckling mice were isolated, overlapped fragments were amplified by RT-PCR and full-length genomes were assembled to analyze the nucleotide and deduced protein similarities and phylogenetic analyses from Chinese Ferret-Badger, dog, sika deer, vole, used vaccine strain were determined. The four full-length genomes were sequenced completely and had the same genetic structure with the length of 11, 923 nts or 11, 925 nts including 58 nts-Leader, 1353 nts-NP, 894 nts-PP, 609 nts-MP, 1575 nts-GP, 6386 nts-LP, and 2, 5, 5 nts- intergenic regions(IGRs), 423 nts-Pseudogene-like sequence (psi), 70 nts-Trailer.

[Sequencing and analyses on glycoprotein gene of rabies viruses isolated in Zhejiang province, China].

Objective: Based on sequencing the genomes of glycoprotein (GP) gene of rabies viruses isolated in Zhejiang, we analyzed the properties of rabies viruses genetic variation in molecular level, and to compare with those of other representative vaccine strains and street virus strains, get the information about rabies viruses variation.

Methods: Suckling mice against rabies virus were selected. Overlapped fragments were amplified by RT-PCR and full-length genomes were assembled to analyze the nucleotide and deduced protein similarities and phylogenetic analyses of the GP genes.

[Complete genome sequencing and analyses of rabies viruses isolated from wild animals (Chinese Ferret-Badger) in Zhejiang province].

Objective: Based on sequencing the full-length genomes of two Chinese Ferret-Badger, we analyzed the properties of rabies viruses genetic variation in molecular level to get information on prevalence and variation of rabies viruses in Zhejiang, and to enrich the genome database of rabies viruses street strains isolated from Chinese wildlife.

Methods: Overlapped fragments were amplified by RT-PCR and full-length genomes were assembled to analyze the nucleotide and deduced protein similarities and phylogenetic analyses of the N genes from Chinese Ferret-Badger, sika deer, vole, dog. Vaccine strains were then determined.

[New animal hosts of rabies virus in mountain areas in Zhejiang province].

Objective: To understand the prevalence of rabies among wild animals and the animal species in rabies epidemic areas of Zhejiang province.

Methods: One hundred and sixty samples were collected from the brain tissues of cats, stoats, Apodemus agrarius, Moschus chinensis, and Sus scrofa in Lishui and Chunan cities of Zhejiang province. Each sample was divided into four parts: cerebrum, mesencephali, cerebellum and gyms hippocampi which were used to determine the positive samples by detection of rabies virus specific antigens and nucleotides, using DFA and RT-PCR methods.

[Study on the difference of genes and the type identification of hantavirus from Lishui, Zhejiang province].

Objective: To isolate hantavirus from Lishui county--one of the epidemic regions for hemorrhagic fever with renal syndrome (HFRS), in Zhejiang province, and to identify the serotype and molecular/biological characteristics of a new HTN subtype hantavirus (HV) strains, hopefully to provide evidence for HFRS prevention and therapy.

Methods: Data on the host animals was collected from Lishui, Zhejiang province in 2007. Direct immunofluorscece assay was adopted to determine HFRS antigens and the lung tissues from HV infected Vero-E6 cells for HV isolation, then total RNA was extracted from Hantavirus Lishui strains and amplified by RT-PCR M, S segments of strains genome were also cloned and sequenced and compared with those of other strains of HV.

Effects of H pylori infection on gap-junctional intercellular communication and proliferation of gastric epithelial cells in vitro.

Aim: To explore the effects of H pylori infection on gap-junctional intercellular communication (GJIC) and proliferation of gastric epithelial cells in vitro.

Methods: A human gastric epithelial cell line (SGC-7901) cultured on coverslips was exposed overnight to intact H pylori (CagA(+) or CagA(-) strains) and sonicated extracts, respectively. GJIC between the cells was detected by fluorescence redistribution after photobleaching (FRAP) technique.