[Establish and optimization of real-time fluorescent reverse transcription loop-mediated isothermal amplification for detection of avian influenza H5 hemagglutinin gene].

H5 subtype avian influenza (AIV-H5) is a major causative agent of animalloimia a rapid and sensitive molecular biological diagnosis is crucial to the control program of AIV-H5. AIV-H5 real-time fluorescent reverse transcription loop-mediated isothermal amplification (qRT-LAMP) was established by means of heat treatment of the samples. The sensitivity, specificity and repeatability of this method were assessed and the performance of Calcein,SYBR Green I,HNB,SYTO 81 in colorimetric detection was comparatively analyzed to screen the optimum dye.

Rapid detection and simultaneous genotyping of Cronobacter spp. (formerly Enterobacter sakazakii) in powdered infant formula using real-time PCR and high resolution melting (HRM) analysis.

Cronobacter spp. is an emerging pathogen that causes meningitis, sepsis, bacteremia, and necrotizing enterocolitis in neonates and children. The present study developed an assay integrating real-time PCR and high resolution melting (HRM) analysis targeting the OmpA gene for the specific detection and rapid identification of Cronobacter spp.

Subtyping animal influenza virus with general multiplex RT-PCR and Liquichip high throughput (GMPLex).

This study developed a multiplex RT-PCR integrated with luminex technology to rapidly subtype simultaneously multiple influenza viruses. Primers and probes were designed to amplify NS and M genes of influenza A viruses HA gene of H1, H3, H5, H7, H9 subtypes, and NA gene of the N1 and N2 subtypes. Universal super primers were introduced to establish a multiplex RT-PCR (GM RT-PCR).

[Detection of bovine, goat, pig and chicken derived ingredients in animal products with universal PCR-microarray method].

We analyzed the sequence of vertebrate molecular marker genes, then we selected the mitochondrial DNA (mtDNA) 16S rRNA gene as marker gene. In order to detect four kinds of animal-derived ingredients, which including bovine, goat, pig and chicken. We utilized a pair of universal primers, designed four sets of species-specific microarray probes and two pairs of quality control probes.

Rapid determination of clenbuterol in urine by a competitive bead immunoassay based on Luminex technology.

A new competitive bead immunoassay (CBIA) based on Luminex technology for detecting clenbuterol in urine was reported. The carboxylated fluorescent beads were directly coated with clenbuterol derivatives without carrier protein spacer. Clenbuterol antibody was biotinylated, which was used for clenbuterol detection in combination with the functionalized bead and streptavidin-phycoerythrin (SAPE).

A rapid assay for detecting antibody against Bluetongue virus with a latex agglutination test using recombinant VP7 antigen.

A latex agglutination test (LAT) for detecting antibody against Bluetongue virus (BTV) in ruminants was developed using latex beads coupled with recombinant VP7 protein. Compared with competitive enzyme-linked immunosorbent assay (ELISA), the specificity and sensitivity of the LAT were 99.0% and 93.

Development and evaluation of an immunochromatographic strip for the detection of serum antibodies against bluetongue virus.

In this study, an immunochromatographic strip (ICS) was developed for the detection of bluetongue virus (BTV) serum antibodies. Colloidal gold particles labeled with streptococcal protein G (SPG), which can bind to the F(C) fragment of mammalian immunoglobulins, were used as the detector reagent. A recombinant VP7 BTV protein and a purified rabbit anti-SPG antibody were immobilized on test and control regions of a nitrocellulose membrane, respectively.

[Multiplex fluorescent real-time PCR detection of bovine, goat and sheep derived materials in animal products].

We designed the specific primers and TaqMan probes targeting cytochrome b genes of mitochondrial DNA from bovine, goat and sheep. We used different fluorescents to label the probes. After optimization of reaction conditions, we set up a multiplex fluorescent real-time PCR method to detect bovine, goat and sheep derived materials, simultaneously.