Hua Xiu-guo

Proteomic study of Mycoplasma suis using the gel-based shotgun strategy.

Mycoplasma suis (M. suis) is an uncultivable pathogen that colonizes the surface of porcine erythrocytes. In the present study, gel-based LC-MS/MS was performed to analyze the proteomic composition of M. suis. We identified 191 proteins with two or more peptides and additional 217 putative proteins with one peptide hit by cross-species searching the Mollicutes protein databases. Resultant proteins were classified based on their molecular functions. The majority of enzymes involved in central carbon metabolic pathways and nucleotide biosynthesis were confirmed by searching the KEGG database. The present study is the first report addressing the proteome of M. suis. Results indicate that the energy source of M. suis might depend mainly on glycometabolism.

Overexpression of complement receptor type I (CR1, CD35) on erythrocytes in patients with hemoplasma infection.

In the present study, 12 patients with fever of undetermined origin, anemia and icterus were diagnosed with hemoplasma infection by light microscopy, transmission electron microscopy examination and PCR assay after being excluded from other usual febrile diseases. Complement receptor type I (CR1, CD35) expression on the surface of erythrocytes was assessed by flow cytometry using mouse anti‐human CD35 antibody. Compared with healthy volunteers, the level of CD35 was significantly elevated in patients with severe hemoplasma infection at diagnosis, and decreased sharply after treatment. However, in latent infection cases without clinical manifestations, CD35 expression showed an ascending trend but had no statistical difference compared to the healthy controls. The present study demonstrated that hemoplasma infection can induce high levels of expression of CR1 on the membrane of red blood cells, which may be a reaction to the immunity challenge.

Development and application of a TaqMan-MGB real-time RT-PCR assay for the detection of porcine epidemic diarrhoea virus strains in China

Abstract Introduction: A real-time RT-PCR method for identification and quantification of porcine epidemic diarrhoea virus (PEDV) strains in China was developed. Material and Methods: Based on the conserved sequence of the PEDV nucleocapsid (N) gene, a primer pair and probe were designed to establish a TaqMan-MGB real-time RT-PCR assay for quantitative detection of the virus. The sequence was cloned into the pMD18-T vector and a series of diluted recombinant plasmids were used to generate a standard curve with an R2 value of 0.999. Results: The developed quantitative PCR assay detected viral titres as low as 0.1 TCID50 with high specificity and no cross-reaction with other porcine viruses (PoRV, TGEV, PRRSV, or CSFV). The intra-batch and inter-batch coefficients of variation were both less than 1%, which indicated good reproducibility. Thirty clinical diarrhoea samples obtained from pigs in Shanghai and Fujian were analysed using this quantitative PCR assay. Out of these samples, 93.3% were found to be PEDV positive. Conclusion: This approach is suitable for clinical sample identification and pathogenesis studies.

Bartonella henselae infection and its effects on human health

The genus Bartonella consists of 20 species and subspecies, and of these, 10 species are responsible for human infections. The most common bartonellosis cat scratch disease (Bartonella henselae), Carrions disease (Bartonella bacilliformis), and trench fever (Bartonella quintana) have been recorded worldwide. Bartonella henselae is the most common Bartonella infection worldwide. As diagnostic techniques improve, the clinical spectrum resulting from B. henselae infection is widening. The microbiological features, clinical manifestations, diagnosis, and treatment will be reviewed.