Development of an ELISA based on a multi-fragment antigen of infectious bronchitis virus for antibodies detection.

Objectives: To develop a cost-effective ELISA for detection of antibodies against infectious bronchitis virus (IBV) by using a multi-fragment protein as coating antigen.

Results: A multi-fragment antigen, termed BE, which was composed of eight antigenic fragments selected from the three major proteins (S, M, and N) of IBV, was expressed in Escherichia coli. The entire protein had a molecular weight of 61.

Development of a multi-epitope antigen of S protein-based ELISA for antibodies detection against infectious bronchitis virus.

An indirect enzyme-linked immunosorbent assay (ELISA) method based on a novel multi-epitope antigen of S protein (SE) was developed for antibodies detection against infectious bronchitis virus (IBV). The multi-epitope antigen SE protein was designed by arranging three S gene fragments (166-247 aa, S1 gene; 501-515 aa, S1 gene; 8-30 aa, S2 gene) in tandem. It was identified to be approximately 32 kDa as a His-tagged fusion protein and can bind IBV positive serum by western blot analysis.

Comparative dynamic distribution of avian infectious bronchitis virus M41, H120, and SAIBK strains by quantitative real-time RT-PCR in SPF chickens.

Avian infectious bronchitis is an acute, highly contagious disease of chickens. To study the differences of dynamic distribution between nephropathogenic infectious bronchitis virus (IBV) strains such as SAIBK and other strains (the M41 and H120 strains), relative quantitative real-time reverse transcription-polymerase chain reaction was developed by housekeeping gene selection. Glyceraldehyde-3-phosphate dehydrogenase and Ubiquitin were chosen for normalization in this experimental set.

Establishment of reverse genetics system for infectious bronchitis virus attenuated vaccine strain H120.

Infectious bronchitis virus (IBV) strain H120 was successfully rescued as infectious clone by reverse genetics. Thirteen 1.5-2.

Detection of clinically important β-lactamases in commensal Escherichia coli of human and swine origin in western China.

Data correlating β-lactamases found in commensal Escherichia coli of human and animal origin are limited. In this study, 447 commensal E. coli isolates from the faeces of humans and swine (280 human isolates from four hospitals and 167 swine isolates from seven farms) were collected between September 2006 and January 2009 in western China.

Complete genome sequence and recombination analysis of infectious bronchitis virus attenuated vaccine strain H120.

The strain H120 of infectious bronchitis virus (IBV) is one of the earliest and representative attenuated live Infectious Bronchitis vaccine strains. To investigate the genomic feature of H120 and further understand its role in the epidemiology of IBV, complete genome of H120 was sequenced and compared with sequences of other IBV strains by phylogenetic and recombination analysis. The complete genome of H120 is 27631 nucleotides in length and has a similar structure with that of Beaudette strain.

An effective method for isolation of DNA from pig faeces and comparison of five different methods.

Polymerase chain reaction (PCR) detection of microorganism in faecal specimens is hampered by poor recovery of DNA and by the presence of PCR inhibitors. In this paper, we describe a new modified method for extracting PCR-quality microbial community DNA from pig faecal samples, which combines the pretreatment with polyformaldehyde, and subsequent DNA lysis in the presence of CTAB, salt, PVP, and beta-mercaptoethanol, followed by isolation of nucleic acids using chloroform (no phenol) based protocol. The method resulted in a 1.